Anti-GPRC5d monoclonal antibodies and uses thereof

Abstract

Provided are antibodies or fragment thereof having binding specificity to the human GPRC5D protein. The antibodies or fragment thereof are capable of targeting cancer cells expressing GPRC5D, and thus can be used to treat the cancer, in particular hematological cancer.

Description

(19) *EP004670724A2* (11) EP 4 670 724 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: 31.12.2025 Bulletin 2026 / 01 (21) Application number: 25209453.7 (22) Date of filing: 05.01.2022 (51) International Patent Classification (IPC): A61K 35 / 15 (2025.01) (52) Cooperative Patent Classification (CPC): A61P 35 / 00; A61K 35 / 15; A61K 35 / 17; A61K 47 / 68031; A61K 47 / 6849; C07K 16 / 28; A61K 2039 / 505; A61K 2039 / 54; A61K 2039 / 545; C07K 2317 / 24; C07K 2317 / 56; C07K 2317 / 565; C07K 2317 / 732; C07K 2317 / 76; C07K 2317 / 77; (Cont.) (84) Designated Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR Designated Extension States: BA ME Designated Validation States: MA TN (30) Priority: 05.01.2021 PCT / CN2021 / 070314 (62) Document number(s) of the earlier application(s) in accordance with Art. 76 EPC: 22736532.7 / 4 274 851 (71) Applicant: Lanova Medicines Development Co., Ltd. Shanghai 201315 (CN) (72) Inventors: • LI, Runsheng Shanghai 201315 (CN) • HUANG, Wentao Shanghai 201315 (CN) (74) Representative: AstraZeneca Intellectual Property Eastbrook House Shaftesbury Road Cambridge CB2 8BF (GB) Remarks: •The complete document including Reference Table(s) and the Sequence Listing(s) can be downloaded from the EPO website •This application was filed on 17.10.2025 as a divisional application to the application mentioned under INID code 62. (54) ANTI‑GPRC5D MONOCLONAL ANTIBODIES AND USES THEREOF (57) Provided are antibodies or fragment thereof havingbinding specificity to the humanGPRC5Dprotein. The antibodies or fragment thereof are capable of target- ing cancer cells expressing GPRC5D, and thus can be used to treat the cancer, in particular hematological can- cer. EP 4 67 0 72 4 A 2 Processed by Luminess, 75001 PARIS (FR) (Cont. next page) (52) Cooperative Patent Classification (CPC): (Cont.) C07K 2317 / 92 2 EP 4 670 724 A2 Description BACKGROUND

[0001] G-protein-coupled receptor family C group 5memberD (GPRC5D) ismember of theG protein-coupled receptor family. GPRC5D is a transmembrane protein. Overexpression of GPRC5D was observed in patients with multiple myeloma. In particular, its high expression had a significant correlation with poor outcome of the disease and treatment.

[0002] Given its specific high expression on malignant cells, it has been proposed that antibodies that are specific to GPRC5D can be useful for treating the malignancy, such as via a bispecific T-cell-redirecting antibody, or through antibody-dependent cellular cytotoxicity (ADCC). SUMMARY

[0003] Anti-GPRC5D antibodies, including their humanized derivatives, are discovered herein that have high affinity to the human GPRC5D protein. The antibodies or fragment thereof are capable of targeting cancer cells expressing GPRC5D, and thus can be used to treat the cancer, in particular hematological cancer.

[0004] One embodiment of the present disclosure provides an antibody or antigen-binding fragment thereof having binding specificity to a human G-protein-coupled receptor family C group 5 member D (GPRC5D) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region (VH) comprising heavy chain complementarity determining regions CDRH1, CDRH2, and CDRH3 and a light chain variable region (VL) comprising complementarity determining regions CDRL1, CDRL2, and CDRL3. In some embodiments, the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise: (a) the amino acid sequences of SEQ ID NO:29‑34; (b) the amino acid sequences of SEQ ID NO:42‑47; (c) the amino acid sequences of SEQ ID NO:54‑59; or (d) the amino acid sequences of SEQ ID NO:68‑73.

[0005] In some embodiments, the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:29‑34. In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:7 and 35‑37, and the VL comprises an amino acid sequence selected from the groupconsistingofSEQ IDNO:8and38‑41. In someembodiments, theVHcomprises theaminoacid sequenceofSEQ ID NO:35, and the VL comprises the amino acid sequence of SEQ ID NO:38.

[0006] In some embodiments, the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:42‑47. In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:9 and 48‑50, and the VL comprises an amino acid sequence selected from the group consisting of SEQ IDNO:10 and 51‑53. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO:48, and the VL comprises the amino acid sequence of SEQ ID NO:51.

[0007] In some embodiments, the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:54‑59. In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:61‑64, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:16 and 65‑67. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO:61, and the VL comprises the amino acid sequence of SEQ ID NO:65. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO:63, and the VL comprises the amino acid sequence of SEQ ID NO:65.

[0008] In some embodiments, the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:68‑73. In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1 and 74‑79, and the VL comprises an amino acid sequence selected from the groupconsistingofSEQ IDNO:2and80‑86. In someembodiments, theVHcomprises theaminoacid sequenceofSEQ ID NO:76, and the VL comprises the amino acid sequence of SEQ ID NO:82. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO:77, and the VL comprises the amino acid sequence of SEQ ID NO:82.

[0009] In some embodiments, the antibody or fragment thereof is humanized. In some embodiments, the antibody or fragment thereof is ADCC-competent.

[0010] Also provided, in some embodiments, the antibody or fragment thereof further comprises a cytotoxic drug conjugated to the antibody or fragment thereof.

[0011] Also provided is a bispecific antibody, comprising the antigen-binding fragment of the present disclosure, and a second antigen-binding fragment having specificity to a second target protein. In some embodiments, the second target protein is selected from the group consisting of CD3, CD16, CD19, CD28, CD64 and 4‑1BB. In some embodiments, the second target protein is CD3. In some embodiments, the second target protein is 4‑1BB.

[0012] Also provided is a method of treating cancer in a patient in need thereof, comprising administering to the patient the antibody or fragment thereof of the present disclosure. Also provided is amethod of treating cancer in a patient in need thereof, comprising (a) treating a Tcell, a natural killer (NK) cell, or a macrophage, in vitro, with the antibody or fragment thereof of the present disclosure, and (b) administering the treated cell to the patient. 3 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55

[0013] In some embodiments, the cancer is a hematological cancer, such as aGPRC5D-expressing B cell cancer (e.g., multiple myeloma). BRIEF DESCRIPTION OF THE DRAWINGS

[0014] FIG. 1 shows binding ofmurine antibodies collected fromhybridoma supernatant toGPRC5Dexpressed onCHO-K1 cells. FIG. 2 shows binding of chimeric antibodies to GPRC5D expressed on CHO-K1 cells. FIG. 3 shows that antibody binding induced endocytosis. FIG. 4 shows that the antibodies were ADCC-competent. FIG. 5‑8 show the affinity testing results of the humanized antibodes derived from 6G10D9, 58F9G10, 34D3H1, and 37B9C4, respectively. FIG. 9 confirms the binding of selected humanized antibodies to GPRC5D expressed on CHO-K1 cells. FIG. 10 confirms the binding of selected humanized antibodies to GPRC5D expressed on NCI-H929 cells. FIG. 11 shows the ADCC efficacy of tested humanized antibodies. FIG. 12 shows the internalization induction activity of the tested humanized antibodies. FIG. 13 shows the cell killing activities of the tested antibody-drug conjugates. FIG. 14 shows the tumor inhibition activities of the tested antibody. DETAILED DESCRIPTION Definitions

[0015] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "an antibody," is understood to represent one or more antibodies. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0016] As used herein, the term "polypeptide" is intended to encompass a singular "polypeptide" as well as plural "polypeptides," and refers toamolecule composedofmonomers (aminoacids) linearly linkedbyamidebonds (alsoknown as peptide bonds). The term "polypeptide" refers to any chain or chains of two ormore amino acids, and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, "protein," "amino acid chain," or any other term used to refer to a chain or chains of two ormore amino acids, are includedwithin the definition of "polypeptide," and the term "polypeptide" may be used instead of, or interchangeably with any of these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of the polypeptide, including without limitation glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, or modification by non‑ naturally occurring amino acids. A polypeptidemay be derived from a natural biological sourceor producedby recombinant technology, but is not necessarily translated fromadesignatednucleic acid sequence. It may be generated in any manner, including by chemical synthesis.

[0017] "Homology" or "identity" or "similarity" refers to sequence similarity between twopeptides or between twonucleic acid molecules. Homology can be determined by comparing a position in each sequence which may be aligned for purposesof comparison.Whenaposition in the compared sequence is occupiedby the samebaseor aminoacid, then the molecules are homologous at that position. A degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. An "unrelated" or "non-homologous" sequence shares less than 40% identity, though preferably less than 25% identity, with one of the sequences of the present disclosure.

[0018] A polynucleotide or polynucleotide region (or a polypeptide or polypeptide region) has a certain percentage (for example, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 98 % or 99 %) of "sequence identity" to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences. 4 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55

[0019] The term "an equivalent nucleic acid or polynucleotide" refers to a nucleic acid having a nucleotide sequence havinga certain degreeof homology, or sequence identity,with thenucleotide sequenceof the nucleic acid or complement thereof. A homolog of a double stranded nucleic acid is intended to include nucleic acids having a nucleotide sequence which has a certain degree of homologywith or with the complement thereof. In one aspect, homologs of nucleic acids are capable of hybridizing to the nucleic acid or complement thereof. Likewise, "an equivalent polypeptide" refers to a polypeptide having a certain degree of homology, or sequence identity, with the amino acid sequence of a reference polypeptide. In some aspects, the sequence identity is at least about 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%. In someaspects, the equivalent polypeptide or polynucleotide has one, two, three, four or five addition, deletion, substitution and their combinations thereof as compared to the reference polypeptide or polynucleotide. In some aspects, the equivalent sequence retains the activity (e.g., epitope-binding) or structure (e.g., salt-bridge) of the reference sequence.

[0020] As used herein, an "antibody" or "antigen-binding polypeptide" refers to a polypeptide or a polypeptide complex that specifically recognizes and binds to an antigen. An antibody can be a whole antibody and any antigen binding fragment or a single chain thereof. Thus the term "antibody" includes any protein or peptide containing molecule that comprisesat least aportionof an immunoglobulinmoleculehavingbiological activity of binding to theantigen.Examplesof such include, but arenot limited to acomplementarity determining region (CDR)of a heavyor light chain or a ligandbinding portion thereof, a heavy chain or light chain variable region, a heavy chain or light chain constant region, a framework (FR) region, or any portion thereof, or at least one portion of a binding protein.

[0021] The terms "antibody fragment" or "antigen-binding fragment", as used herein, is a portion of an antibody such as F(ab’)2, F(ab)2, Fab’, Fab, Fv, scFvand the like.Regardlessof structure, anantibody fragment bindswith the sameantigen that is recognized by the intact antibody. The term "antibody fragment" includes aptamers, spiegelmers, and diabodies. The term "antibody fragment" also includes any synthetic or genetically engineered protein that acts like an antibody by binding to a specific antigen to form a complex.

[0022] A "single-chain variable fragment" or "scFv" refers to a fusionprotein of the variable regionsof theheavy (VH) and light chains (VL) of immunoglobulins. In someaspects, the regions are connectedwith a short linker peptide of ten to about 25 amino acids. The linker can be rich in glycine for flexibility, as well as serine or threonine for solubility, and can either connect the N-terminus of the VH with the C-terminus of the VL, or vice versa. This protein retains the specificity of the original immunoglobulin, despite removal of the constant regions and the introduction of the linker. ScFv molecules are known in the art and are described, e.g., in US patent 5,892,019.

[0023] The termantibody encompasses various broad classes of polypeptides that can be distinguished biochemically. Those skilled in the art will appreciate that heavy chains are classified as gamma,mu, alpha, delta, or epsilon (y,µ,α, δ, ε) with some subclasses among them (e.g., γ1‑ γ4). It is the nature of this chain that determines the "class" of the antibody as IgG, IgM, IgA IgG, or IgE, respectively. The immunoglobulin subclasses (isotypes) e.g., IgG1, IgG2, IgG3, IgG4, IgG5, etc. are well characterized and are known to confer functional specialization. Modified versions of each of these classes and isotypes are readily discernable to the skilled artisan in viewof the instant disclosure and, accordingly, arewithin the scope of the instant disclosure. All immunoglobulin classes are clearly within the scope of the present disclosure, the following discussion will generally be directed to the IgG class of immunoglobulin molecules. With regard to IgG, a standard immunoglobulin molecule comprises two identical light chain polypeptides of molecular weight approximately 23,000 Daltons, and two identical heavy chain polypeptides of molecular weight 53,000‑70,000. The four chains are typically joinedbydisulfidebonds ina "Y" configurationwherein the light chainsbracket theheavychainsstartingat themouthof the "Y" and continuing through the variable region.

[0024] Antibodies, antigen-binding polypeptides, variants, or derivatives thereof of the disclosure include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized, primatized, or chimeric antibodies, single chain antibodies, epitope-binding fragments, e.g., Fab, Fab’ and F(ab’)2, Fd, Fvs, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv), fragments comprising either a VK or VH domain, fragments produced by a Fab expression library, and anti‑ idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies to LIGHT antibodies disclosedherein). Immunoglobulin or antibodymoleculesof thedisclosure canbeof any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgAl and IgA2) or subclass of immunoglobulin molecule.

[0025] Light chains are classified as either kappa or lambda (K, λ). Each heavy chain class may be bound with either a kappa or lambda light chain. In general, the light and heavy chains are covalently bonded to each other, and the "tail" portionsof the twoheavychainsarebonded toeachother bycovalent disulfide linkagesor non-covalent linkageswhen the immunoglobulins are generated either by hybridomas, B cells or genetically engineered host cells. In the heavy chain, the amino acid sequences run from anN-terminus at the forked ends of the Y configuration to the C-terminus at the bottom of each chain.

[0026] Both the light and heavy chains are divided into regions of structural and functional homology. The terms "constant" and "variable" are used functionally. In this regard, it will be appreciated that the variable domains of both the light (VK) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light chain (CK) and the heavy chain (CH1, CH2 or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, and the like. By convention the numbering of the 5 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 constant region domains increases as they become more distal from the antigen-binding site or amino‑ terminus of the antibody. The N-terminal portion is a variable region and at the C-terminal portion is a constant region; the CH3 and CK domains actually comprise the carboxy-terminus of the heavy and light chain, respectively.

[0027] As indicatedabove, thevariable regionallows theantibody to selectively recognizeandspecifically bindepitopes on antigens. That is, the VK domain and VH domain, or subset of the complementarity determining regions (CDRs), of an antibody combine to form the variable region that defines a three dimensional antigen-binding site. This quaternary antibody structure forms the antigen-binding site present at the end of each arm of the Y. More specifically, the antigen- binding site is definedby threeCDRsoneachof theVHandVKchains (i.e.CDR-H1,CDR-H2,CDR-H3,CDR-L1,CDR-L2 and CDR-L3). In some instances, e.g., certain immunoglobulin molecules derived from camelid species or engineered based on camelid immunoglobulins, a complete immunoglobulin moleculemay consist of heavy chains only, with no light chains. See, e.g., Hamers-Casterman et al., Nature 363:446‑448 (1993).

[0028] In naturally occurring antibodies, the six "complementarity determining regions" or "CDRs" present in each antigen-binding domain are short, non-contiguous sequences of amino acids that are specifically positioned to form the antigen-binding domain as the antibody assumes its three dimensional configuration in an aqueous environment. The remainder of the amino acids in the antigen-binding domains, referred to as "framework" regions, show less inter- molecular variability. The framework regions largely adopt a β-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the β -sheet structure. Thus, framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions. The antigen-binding domain formed by the positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the non-covalent binding of the antibody to its cognate epitope. The amino acids comprising the CDRs and the framework regions, respectively, can be readily identified for any given heavy or light chain variable region by one of ordinary skill in the art, since they have been precisely defined (see "Sequences of Proteins of Immunological Interest," Kabat, E., et al., U.S. Department of Health andHumanServices, (1983); andChothia and Lesk, J. Mol. Biol., 196:901‑917 (1987)).

[0029] In the case where there are two or more definitions of a term which is used and / or accepted within the art, the definition of the term as used herein is intended to include all such meanings unless explicitly stated to the contrary. A specific example is the use of the term "complementarity determining region" ("CDR") to describe the non-contiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. This particular region has been described by Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983) and byChothia et al., J.Mol. Biol. 196:901‑917 (1987), which are incorporated herein by reference in their entireties. The CDR definitions according to Kabat and Chothia include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The appropriate amino acid residueswhich encompass theCDRs as defined by each of the above cited references are set forth in the table belowas a comparison.Theexact residuenumberswhichencompassaparticularCDRwill varydependingon thesequenceandsize of the CDR. Those skilled in the art can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of the antibody. Kabat Chothia CDR-H1 31‑35 26‑32 CDR-H2 50‑65 52‑58 CDR-H3 95‑102 95‑102 CDR-L1 24‑34 26‑32 CDR-L2 50‑56 50‑52 CDR-L3 89‑97 91‑96

[0030] Kabat et al. also defined a numbering system for variable domain sequences that is applicable to any antibody. One of ordinary skill in the art can unambiguously assign this system of "Kabat numbering" to any variable domain sequence, without reliance on any experimental data beyond the sequence itself. As used herein, "Kabat numbering" refers to the numbering system set forth by Kabat et al., U.S. Dept. of Health andHumanServices, "Sequence of Proteins of Immunological Interest" (1983).

[0031] In addition to table above, the Kabat number system describes the CDR regions as follows: CDR-H1 begins at approximately amino acid 31 (i.e., approximately 9 residues after the first cysteine residue), includes approximately 5‑7 amino acids, and ends at the next tryptophan residue. CDR-H2 begins at the fifteenth residue after the end of CDR-H1, 6 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 includes approximately 16‑19 amino acids, and ends at the next arginine or lysine residue. CDR-H3 begins at approximately the thirty third amino acid residue after the end of CDR-H2; includes 3‑25 amino acids; and ends at the sequence W-G-X-G, where X is any amino acid. CDR-L1 begins at approximately residue 24 (i.e., following a cysteine residue); includes approximately 10‑17 residues; and ends at the next tryptophan residue. CDR-L2 begins at approxi- mately the sixteenth residue after the end of CDR-L1 and includes approximately 7 residues. CDR-L3 begins at approximately the thirty third residue after the end of CDR-L2 (i.e., following a cysteine residue); includes approximately 7‑11 residues and ends at the sequence F or W-G-X-G, where X is any amino acid.

[0032] Antibodies disclosed herein may be from any animal origin including birds and mammals. Preferably, the antibodies are human, murine, donkey, rabbit, goat, guinea pig, camel, llama, horse, or chicken antibodies. In another embodiment, the variable region may be condricthoid in origin (e.g., from sharks).

[0033] As used herein, the term "heavy chain constant region" includes amino acid sequences derived from an immunoglobulin heavy chain. A polypeptide comprising a heavy chain constant region comprises at least one of: a CH1domain, a hinge (e.g., upper,middle, and / or lower hinge region) domain, aCH2domain, aCH3domain, or a variant or fragment thereof. For example, an antigen-binding polypeptide for use in thedisclosuremay comprise apolypeptide chain comprising aCH1 domain; a polypeptide chain comprising aCH1 domain, at least a portion of a hinge domain, and aCH2 domain; a polypeptide chain comprising aCH1domain andaCH3domain; a polypeptide chain comprising aCH1domain, at least a portion of a hinge domain, andaCH3domain, or a polypeptide chain comprising aCH1domain, at least a portion of a hingedomain, aCH2domain, andaCH3domain. In another embodiment, a polypeptide of the disclosure comprises a polypeptide chain comprising a CH3 domain. Further, an antibody for use in the disclosuremay lack at least a portion of a CH2domain (e.g., all or part of aCH2domain). As set forth above, it will be understoodbyoneof ordinary skill in the art that the heavy chain constant regionmay bemodified such that they vary in amino acid sequence from the naturally occurring immunoglobulin molecule.

[0034] The heavy chain constant region of an antibody disclosed herein may be derived from different immunoglobulin molecules.Forexample, aheavychainconstant regionofapolypeptidemaycompriseaCH1domainderived froman IgG1 molecule and a hinge region derived from an IgG3 molecule. In another example, a heavy chain constant region can comprise a hinge region derived, in part, from an IgGmolecule and, in part, from an IgG3molecule. In another example, a heavy chain portion can comprise a chimeric hinge derived, in part, from an IgG1 molecule and, in part, from an IgG4 molecule.

[0035] Asused herein, the term "light chain constant region" includes amino acid sequences derived fromantibody light chain. Preferably, the light chain constant region comprises at least one of a constant kappa domain or constant lambda domain.

[0036] A "light chain-heavy chain pair" refers to the collection of a light chain and heavy chain that can form a dimer through a disulfide bond between the CL domain of the light chain and the CH1 domain of the heavy chain.

[0037] Aspreviously indicated, the subunit structures and three dimensional configuration of the constant regions of the various immunoglobulin classes are well known. As used herein, the term "VH domain" includes the amino terminal variable domain of an immunoglobulin heavy chain and the term "CH1 domain" includes the first (most amino terminal) constant region domain of an immunoglobulin heavy chain. The CH1 domain is adjacent to the VH domain and is amino terminal to the hinge region of an immunoglobulin heavy chain molecule.

[0038] As used herein the term "CH2 domain" includes the portion of a heavy chain molecule that extends, e.g., from about residue 244 to residue 360 of an antibody using conventional numbering schemes (residues 244 to 360, Kabat numbering system; and residues 231‑340, EU numbering system; see Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983). The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgGmolecule. It is alsowell documented that theCH3domain extends from theCH2domain to the C-terminal of the IgG molecule and comprises approximately 108 residues.

[0039] As used herein, the term "hinge region" includes the portion of a heavy chainmolecule that joins theCH1domain to theCH2domain.Thishinge regioncomprisesapproximately25 residuesand is flexible, thusallowing the twoN-terminal antigen-binding regions to move independently. Hinge regions can be subdivided into three distinct domains: upper, middle, and lower hinge domains (Roux et al., J. Immunol 161:4083 (1998)).

[0040] Asusedherein the term "disulfidebond" includes thecovalent bond formedbetween twosulfur atoms.Theamino acid cysteine comprises a thiol group that can form a disulfide bond or bridge with a second thiol group. In most naturally occurring IgGmolecules, theCH1andCKregionsare linkedbyadisulfidebondand the twoheavychainsare linkedby two disulfide bonds at positions corresponding to 239 and 242 using the Kabat numbering system (position 226 or 229, EU numbering system).

[0041] As used herein, the term "chimeric antibody" will be held to mean any antibody wherein the immunoreactive region or site is obtained or derived from a first species and the constant region (whichmay be intact, partial or modified in accordance with the instant disclosure) is obtained from a second species. In certain embodiments the target binding region or site will be from a non-human source (e.g. mouse or primate) and the constant region is human. 7 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55

[0042] As used herein, "percent humanization" is calculated by determining the number of framework amino acid differences (i.e., non-CDRdifference) between the humanized domain and the germline domain, subtracting that number from the total number of amino acids, and then dividing that by the total number of amino acids and multiplying by 100.

[0043] By "specifically binds" or "has specificity to," it is generally meant that an antibody binds to an epitope via its antigen-binding domain, and that the binding entails some complementarity between the antigen-binding domain and the epitope. According to this definition, an antibody is said to "specifically bind" to an epitopewhen it binds to that epitope, via its antigen-binding domain more readily than it would bind to a random, unrelated epitope. The term "specificity" is used herein toqualify the relativeaffinitybywhichacertainantibodybinds toacertainepitope.Forexample,antibody "A"maybe deemed to haveahigher specificity for a givenepitope than antibody "B," or antibody "A"maybe said to bind to epitope "C" with a higher specificity than it has for related epitope "D."

[0044] As used herein, the terms "treat" or "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, suchas theprogressionof cancer. Beneficial or desired clinical results include, but arenot limited to, alleviationof symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can alsomean prolonging survival as compared to expected survival if not receiving treatment. Those in needof treatment include thosealreadywith the condition or disorder aswell as thoseprone tohave the condition or disorder or those in which the condition or disorder is to be prevented.

[0045] By "subject" or "individual" or "animal" or "patient" or "mammal," is meant any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired. Mammalian subjects include humans, domestic animals, farm animals, and zoo, sport, or pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows, and so on.

[0046] As used herein, phrases such as "to a patient in need of treatment" or "a subject in need of treatment" includes subjects, suchasmammalian subjects, thatwould benefit fromadministration of anantibody or composition of the present disclosure used, e.g., for detection, for a diagnostic procedure and / or for treatment. Anti-GPRC5D Antibodies

[0047] Using the hybridoma technology, the accompanying experimental examples showed that a number of murine antibodies were obtained. Fourteen of the murine antibodies were sequenced, and humanized antibodies were made. These humanized antibodies exhibited potent GPRC5D-binding activities, and were able to induce receptor-mediated endocytosis. In vivo testing showed that theseantibodieswereactive in inducingADCCand inhibiting tumordevelopment.

[0048] Four of the murine antibodies, 34D3H1, 37B9C4, 58F9G10 and 6G10D9, went through the humanization process. Some of the humanized antibodies, including 6-H3L3, 6-H4L3 (both derived from 6G10D9), 58-H1L1, 58-H3L1 (both derived from 58F9G10), 34-H1L1 (derived from 34D3H1) and 37-H1L1 (derived from 37B9C4) further showed promise to continued clinical development.

[0049] In accordance with one embodiment of the present disclosure, therefore, provided is an antibody or antigen- binding fragment thereof having binding specificity to a human G-protein-coupled receptor family C group 5 member D (GPRC5D) protein. The antibody or fragment thereof includes a heavy chain variable region (VH) comprising heavy chain complementarity determining regions CDRH1, CDRH2, and CDRH3 and a light chain variable region (VL) comprising complementarity determining regions CDRL1, CDRL2, and CDRL3.

[0050] In someembodiments, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2, andCDRL3, respectively, include (a) the amino acid sequences of SEQ ID NO: 29‑34; (b) the amino acid sequences of SEQ ID NO: 42‑47; (c) the amino acid sequences of SEQ IDNO:54‑59, or SEQ IDNO:54, 60, and 56‑59; or (d) the amino acid sequences of SEQ IDNO:68‑73.

[0051] Inoneembodiment, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2,andCDRL3, respectively, include theamino acid sequences of SEQ ID NO:29‑34. Example sequences for the VH include SEQ ID NO:7 and 35‑37 or a sequence having at least 85%, 90%, or 95% sequence identity to any of SEQ ID NO:7 and 35‑37. Example sequences for the VL includeSEQ IDNO:8 and38‑41or a sequence having at least 85%, 90%, or 95%sequence identity to any of SEQ IDNO:8 and 38‑41.

[0052] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:35 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:35. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:38 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:38.

[0053] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:35 and a VL that includes the amino acid sequence of SEQ ID NO:38.

[0054] Inoneembodiment, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2,andCDRL3, respectively, include theamino acid sequences of SEQ ID NO:42‑47. Example sequences for the VH include SEQ ID NO: 9 and 48‑50 or a sequence 8 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 having at least 85%, 90%, or 95% sequence identity to any of SEQ ID NO: 9 and 48‑50. Example sequences for the VL includeSEQ IDNO:10 and51‑53or a sequencehaving at least 85%, 90%, or 95%sequence identity to any of SEQ IDNO: 10 and 51‑53.

[0055] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:48 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:48. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:51 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:51.

[0056] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:48 and a VL that includes the amino acid sequence of SEQ ID NO:51.

[0057] Inoneembodiment, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2,andCDRL3, respectively, include theamino acid sequences of SEQ IDNO:54‑59. Example sequences for the VH include SEQ IDNO: 61‑64 or a sequence having at least 85%,90%,or 95%sequence identity toanyofSEQ IDNO:61‑64.Example sequences for theVL includeSEQ IDNO: 16 and 65‑67 or a sequence having at least 85%, 90%, or 95% sequence identity to any of SEQ ID NO: 16 and 65‑67.

[0058] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:61 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:61. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:65 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:65.

[0059] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:61 and a VL that includes the amino acid sequence of SEQ ID NO:65.

[0060] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:63 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:63. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:65 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:65.

[0061] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:63 and a VL that includes the amino acid sequence of SEQ ID NO:65.

[0062] Inoneembodiment, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2,andCDRL3, respectively, include theamino acid sequences of SEQ ID NO:54, 60, and 56‑59. Example sequences for the VH include SEQ ID NO:15 or a sequence having at least 85%, 90%, or 95% sequence identity to any of SEQ IDNO15. Example sequences for the VL include SEQ ID NO: 16 and 65‑67 or a sequence having at least 85%, 90%, or 95% sequence identity to any of SEQ ID NO: 16 and 65‑67.

[0063] Inoneembodiment, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2,andCDRL3, respectively, include theamino acid sequences of SEQ ID NO: 68‑73. Example sequences for the VH include SEQ ID NO: 1 and 74‑79 or a sequence having at least 85%, 90%, or 95% sequence identity to any of SEQ ID NO: 1 and 74‑79. Example sequences for the VL includeSEQ IDNO:2and80‑86or asequencehavingat least 85%,90%,or 95%sequence identity toanyofSEQ IDNO:2 and 80‑86.

[0064] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:76 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:76. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:82 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:82.

[0065] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:76 and a VL that includes the amino acid sequence of SEQ ID NO:82.

[0066] In one embodiment, the VH includes the amino acid sequence of SEQ ID NO:77 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:77. In one embodiment, the VL includes the amino acid sequence of SEQ ID NO:82 or a sequence having at least 85%, 90%, or 95% sequence identity to SEQ ID NO:82.

[0067] Alsoprovided, in someembodiments,areantibodiesandantigen-binding fragments thereof that bind to thesame epitopeonFAPαasanyoneof theseantibodies.Alsoprovided, in someembodiments, areantibodiesandantigen-binding fragments thereof that compete with any one of these antibodies in binding to FAPα, such as one that has a VH that includes the amino acid sequence of SEQ ID NO:77 and a VL that includes the amino acid sequence of SEQ ID NO:82.

[0068] In someembodiments, the antibody or fragment thereof isADCC-competent.Methods andmaterials suitable for making an antibody ADCC-competent are known in the art, such as by using the appropriate Fc fragments or decreasing / removing fucosylation.

[0069] In someembodiments, theCDRH1,CDRH2,CDRH3,CDRL1,CDRL2, andCDRL3, respectively, include (a) the amino acid sequences of SEQ ID NO:29‑34; (b) the amino acid sequences of SEQ ID NO:42‑47; (c) the amino acid sequences of SEQ IDNO:54‑59, or SEQ IDNO:54, 60, and 56‑59; or (d) the amino acid sequences of SEQ IDNO:68‑73, 9 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 wherein each of the recited CDR sequence includes one, two or three conservative amino acid substitution.

[0070] A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residuehavingasimilar side chain. Familiesof aminoacid residueshaving similar side chainshavebeendefined in theart, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a nonessential amino acid residue in an immunoglobulin polypeptide is preferably replacedwith another amino acid residue from the sameside chain family. In another embodiment, a stringof aminoacids canbe replacedwith a structurally similar string that differs in order and / or composition of side chain family members.

[0071] Non-limitingexamplesof conservativeaminoacid substitutionsareprovided in the tablebelow,whereasimilarity score of 0 or higher indicates conservative substitution between the two amino acids. Table B. Conservative Amino Acid Substitutions For Amino Acid Substitution With Alanine D-Ala, Gly, Aib, β-Ala, L-Cys, D-Cys Arginine D-Arg, Lys, D-Lys, Orn D-Orn Asparagine D-Asn, Asp, D-Asp, Glu, D-Glu Gln, D-Gln Aspartic Acid D-Asp, D-Asn, Asn, Glu, D-Glu, Gln, D-Gln Cysteine D-Cys, S-Me-Cys, Met, D-Met, Thr, D-Thr, L-Ser, D-Ser Glutamine D-Gln, Asn, D-Asn, Glu, D-Glu, Asp, D-Asp Glutamic Acid D-Glu, D-Asp, Asp, Asn, D-Asn, Gln, D-Gln Glycine Ala, D-Ala, Pro, D-Pro, Aib, β-Ala Isoleucine D-Ile, Val, D-Val, Leu, D-Leu, Met, D-Met Leucine Val, D-Val, Met, D-Met, D-Ile, D-Leu, Ile Lysine D-Lys, Arg, D-Arg, Orn, D-Orn Methionine D-Met, S-Me-Cys, Ile, D-lle, Leu, D-Leu, Val, D-Val Phenylalanine D-Phe, Tyr, D-Tyr, His, D-His, Trp, D-Trp 10 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) For Amino Acid Substitution With Proline D-Pro Serine D-Ser, Thr, D-Thr, allo-Thr, L-Cys, D-Cys Threonine D-Thr, Ser, D-Ser, allo-Thr, Met, D-Met, Val, D-Val Tyrosine D-Tyr, Phe, D-Phe, His, D-His, Trp, D-Trp Valine D-Val, Leu, D-Leu, Ile, D-Ile, Met, D-Met

[0072] It will also be understood by one of ordinary skill in the art that antibodies as disclosed herein may be modified such that they vary in amino acid sequence from thenaturally occurring binding polypeptide fromwhich theywere derived. For example, a polypeptide or amino acid sequence derived froma designated proteinmay be similar, e.g., have a certain percent identity to the starting sequence, e.g., it may be 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to the starting sequence.

[0073] In someembodiments, theanti-CCR8antibodiesaremodifiedmAbscomprisingamodifiedheavychainconstant region, such as an afucosylated heavy chain, that binds with higher affinity to activating Fcy receptor that mediated enhanced ADCC compared to an unmodified mAb. In some embodiments, the anti-CCR8 antibodies comprises a heavy chainwhich is of a human IgG1 variant that include the single or combination of L234Y, L235Q,G236W,S239D / M, F243L, H268D, D270E, R292P, S298A, Y300L, V305I, K326D, A330L / M, I332E, K334A / E, P396L that enhance ADCC function (all EU numbering).

[0074] In certain embodiments, the antibody comprises an amino acid sequence or one or more moieties not normally associatedwith an antibody. Exemplarymodifications are described inmore detail below. For example, an antibody of the disclosure may comprise a flexible linker sequence, or may be modified to add a functional moiety (e.g., PEG, a drug, a toxin, or a label).

[0075] Antibodies, variants, or derivatives thereof of the disclosure include derivatives that are modified, i.e., by the covalent attachment of any type of molecule to the antibody such that covalent attachment does not prevent the antibody frombinding to theepitope.For example, but not bywayof limitation, theantibodies canbemodified,e.g., byglycosylation, acetylation,pegylation, phosphorylation,phosphorylation,amidation, derivatizationbyknownprotecting / blockinggroups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Additionally, the antibodies may contain one or more non-classical amino acids. Antibody-Drug Conjugates

[0076] GPRC5D is overexpressed on certain malignant hematological cells, such as multiple myeloma cells. The antibodies and fragments of the instant disclosure, therefore, can be used to target those malignant cells for inhibition, inactivation of destruction. In one example, the antibody or fragment is conjugated to an agent that helps to inhibit, inactive or destruct the malignant cell. It is shown herein that the antibodies are able to induce GPRC5D-mediated endocytosis.

[0077] In some embodiments, the antibodies or fragments may be conjugated to therapeutic agents, prodrugs, peptides, proteins, enzymes, viruses, lipids, biological response modifiers, pharmaceutical agents, or PEG. In some embodiments, the conjugatedagentmaybe short interferingRNA (siRNA) or an innatemodulator, suchasaSTimulator of Interferon Genes (STING) agonist, or a TLR7 / 8 agonist.

[0078] In one embodiment, the antibodies or fragments of the disclosure are covalently attached to a drug moiety. The drug moiety may be, or be modified to include, a group reactive with a conjugation point on the antibody. For example, a drug moiety can be attached by alkylation (e.g., at the epsilon-amino group lysines or the N-terminus of antibodies), reductive amination of oxidized carbohydrate, transesterification between hydroxyl and carboxyl groups, amidation at amino groups or carboxyl groups, and conjugation to thiols.

[0079] In some embodiments, the number of drug moieties, p, conjugated per antibody molecule ranges from an averageof 1 to 8; 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In someembodiments, p ranges fromanaverageof 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4 or 2 to 3. In other embodiments, p is an average of 1, 2, 3, 4, 5, 6, 7 or 8. In someembodiments, p ranges fromanaverageofabout1 toabout20,about1 toabout10,about2 toabout10, about2 toabout9, about1 toabout8, about 1 to about 7, about 1 to about 6, about 1 to about 5, about 1 to about 4, about 1 to about 3, or about 1 to about 2. In some embodiments, p ranges fromabout 2 toabout 8, about 2 toabout 7, about 2 toabout 6, about 2 toabout 5, about 2 to about 4 or about 2 to about 3.

[0080] For example, when chemical activation of the protein results in formation of free thiol groups, the protein may be conjugated with a sulfhydryl reactive agent. In one aspect, the agent is one which is substantially specific for free thiol 11 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 groups. Such agents include, for example,malemide, haloacetamides (e.g., iodo, bromoor chloro), haloesters (e.g., iodo, bromoor chloro), halomethyl ketones (e.g., iodo, bromoor chloro), benzylichalides (e.g., iodide,bromideor chloride), vinyl sulfone and pyridylthio.

[0081] Thedrugcanbe linked to theantibodyor fragmentbya linker.Suitable linkers include, for example, cleavableand non-cleavable linkers. A cleavable linker is typically susceptible to cleavage under intracellular conditions. Suitable cleavable linkers include, for example, a peptide linker cleavable by an intracellular protease, such as lysosomal protease or an endosomal protease. In exemplary embodiments, the linker can be a dipeptide linker, such as a valine-citrulline (val- cit), a phenylalanine-lysine (phe-lys) linker, or maleimidocapronic-valine-citruline-p-aminobenzyloxycarbonyl (mc-Val- Cit-PABA) linker. Another linker is Sulfosuccinimidyl‑4‑[N-maleimidomethyl]cyclohexane‑1-carboxylate (smcc). Sulfo- smcc conjugation occurs via a maleimide group which reacts with sulfhydryls (thiols, ‑SH), while its Sulfo-NHS ester is reactive toward primary amines (as found in Lysine and the protein or peptide N-terminus). Yet another linker is maleimidocaproyl (mc). Other suitable linkers include linkers hydrolyzable at a specific pH or a pH range, such as a hydrazone linker. Additional suitable cleavable linkers include disulfide linkers. The linker may be covalently bound to the antibody to suchanextent that theantibodymust bedegraded intracellularly in order for thedrug tobe releasede.g. themc linker and the like.

[0082] A linker can include a group for linkage to the antibody. For example, linker can include an amino, hydroxyl, carboxyl or sulfhydryl reactivegroups (e.g.,malemide,haloacetamides (e.g., iodo,bromoorchloro), haloesters (e.g., iodo, bromoor chloro), halomethyl ketones (e.g., iodo, bromoor chloro), benzylichalides (e.g., iodide,bromideor chloride), vinyl sulfone and pyridylthio).

[0083] In some embodiments, the drug moiety is a cytotoxic or cytostatic agent, an immunosuppressive agent, a radioisotope, a toxin, or the like. The conjugate can be used for inhibiting the multiplication of a tumor cell or cancer cell, causing apoptosis in a tumor or cancer cell, or for treating cancer in a patient. The conjugate can be used accordingly in a variety of settings for the treatment of animal cancers. The conjugate canbe used to deliver a drug to a tumor cell or cancer cell. Without being bound by theory, in some embodiments, the conjugate binds to or associates with a cancer cell expressing GPRC5D, and the conjugate and / or drug can be taken up inside a tumor cell or cancer cell through receptor- mediated endocytosis.

[0084] Once inside the cell, one or more specific peptide sequences within the conjugate (e.g., in a linker) are hydrolytically cleaved by one or more tumor-cell or cancer-cell-associated proteases, resulting in release of the drug. The released drug is then free to migrate within the cell and induce cytotoxic or cytostatic or other activities. In some embodiments, the drug is cleaved from the antibody outside the tumor cell or cancer cell, and the drug subsequently penetrates the cell, or acts at the cell surface.

[0085] Examples of drug moieties or payloads are selected from the group consisting of DM1 (maytansine, N2’- deacetyl-N2’‑(3-mercapto‑1-oxopropyl)‑orN2’-deacetyl-N2’‑(3-mercapto‑1-oxopropyl)‑maytansine),mc-MMAD(6-mal- eimidocaproyl-monomethylauristatin-D or N-methyl-L-valyl-N‑[(1S,2R)‑2-methoxy‑4‑[(2S)‑2‑[(1R,2R)‑1-methoxy‑2- methyl‑3-oxo‑3‑[[(1S)‑2-phenyl‑1‑(2-thiazolyl)ethyl]amino]propyl]‑1-pyr rolidinyl]‑1‑[(1S)‑1-methylpropyl]‑4-oxobu- tyl]‑N-methyl‑(9Cl)‑L-valinamide), mc-MMAF (maleimidocaproyl-monomethylauristatin F or N‑[6‑(2,5-dihydro‑2,5-diox- o‑1H-pyrrol‑1-yl)‑1-oxohexyl]‑N-methyl-L-valyl-L-valyl‑(3R,4S,5S)‑3-methoxy‑5-methyl‑4‑(methylamino)heptanoy- l‑(αR,βR,2S)-β-methoxy-α-methyl‑2-pyrrolidinepropanoyl-L-phenylalanine) andmc-Val-Cit-PABA-MMAE (6-maleimido- caproyl-ValcCit‑(p-aminobenzyloxycarbonyl)‑monomethylauristatin E or N‑[[[4‑[[N‑[6‑(2,5-dihydro‑2,5-dioxo‑1H-pyr- rol‑1-yl)‑1-oxohexyl]‑L-valyl-N5‑(aminocarbonyl)‑L-ornithyl]amino]phenyl]methoxy]carbonyl]‑N-meth yl-L-valyl- N‑[(1S,2R)‑4‑[(2S)‑2‑[(1R,2R)‑3‑[[(1R,2S)‑2-hydroxy‑1-methyl‑2-phenylethyl]amino]‑1-methoxy‑2-methyl‑3-oxopro- pyl]‑1-pyrrolidinyl] ‑2-methoxy‑1‑[(1S)‑1-methylpropyl]‑4-oxobutyl]‑N-methyl-L-valinamide). DM1 is a derivative of the tubulin inhibitor maytansine while MMAD, MMAE, and MMAF are auristatin derivatives. In some embodiments, the drug moiety is selected from the group consisting of mc-MMAF andmc-Val-Cit-PABA-MMAE. In some embodiments, the drug moiety is a maytansinoid or an auristatin.

[0086] The antibodies or fragments may be conjugated or fused to a therapeutic agent, which may include detectable labels such as radioactive labels, an immunomodulator, a hormone, an enzyme, an oligonucleotide, a photoactive therapeutic or diagnostic agent, a cytotoxic agent, whichmay be a drug or a toxin, an ultrasound enhancing agent, a non- radioactive label, a combination thereof and other such agents known in the art.

[0087] The antibodies can be detectably labeled by coupling it to a chemiluminescent compound. The presence of the chemiluminescent-taggedantigen-bindingpolypeptide is thendeterminedbydetecting thepresenceof luminescence that arisesduring thecourseofachemical reaction.Examplesofparticularly useful chemiluminescent labelingcompoundsare luminol, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester.

[0088] The antibodies can also be detectably labeled using fluorescence emittingmetals such as 152Eu, or others of the lanthanide series. These metals can be attached to the antibody using such metal chelating groups as diethylenetria- minepentacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA). Techniques for conjugating various moieties to an antibody are well known, see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243‑56 (Alan R. Liss, Inc. (1985); 12 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 Hellstrom et al., "Antibodies For Drug Delivery", in Controlled Drug Delivery (2nd Ed.), Robinson et al., (eds.), Marcel Dekker, Inc., pp. 623‑ 53 (1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies ’84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475‑506 (1985); "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal AntibodiesForCancerDetectionAndTherapy,Baldwin et al. (eds.), AcademicPress pp. 303‑16 (1985), andThorpeet al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates ", Immunol. Rev. (52:119‑58 (1982)). Bi-functional Molecules and Combination Therapies

[0089] GPRC5D is overexpressed on certain malignant hematological cells, such as multiple myeloma cells. The antibodies and fragments of the instant disclosure, therefore, can be used to target those malignant cells for inhibition, inactivation of destruction. In some embodiments, a bi-functional or bispecific molecule / antibody is provided that targets both the GPRC5D protein and an immune cell.

[0090] In some embodiments, the immune cell is selected from the group consisting of a Tcell, a B cell, a monocyte, a macrophage, a neutrophil, a dendritic cell, a phagocyte, a natural killer cell, an eosinophil, a basophil, and a mast cell. Molecules on the immune cell which can be targeted include, for example, CD3, CD16, CD19, CD28, CD64 and 4‑1BB (also known as CD137). Other examples include PD‑1, CTLA‑4, LAG‑3 (also known as CD223), CD28, CD122, TIM3, OX‑40orOX40L,CD40orCD40L, LIGHT, ICOS / ICOSL,GITR / GITRL, TIGIT,CD27,VISTA,B7H3,B7H4,HEVMorBTLA (also known as CD272), killer-cell immunoglobulin-like receptors (KIRs), and CD47. Specific examples of bispecificity include, without limitation, GPRC5D / CD3.

[0091] Different format of bispecific antibodies are also provided. In some embodiments, each of the anti-PD-L1 fragment and the second fragment each is independently selected from a Fab fragment, a single-chain variable fragment (scFv), or a single-domain antibody. In some embodiments, the bispecific antibody further includes a Fc fragment. Polynucleotides Encoding the Antibodies and Methods of Preparing the Antibodies

[0092] Thepresentdisclosurealsoprovides isolatedpolynucleotidesornucleicacidmoleculesencoding theantibodies, variants or derivatives thereof of the disclosure. The polynucleotides of the present disclosure may encode the entire heavy and light chain variable regions of the antigen-binding polypeptides, variants or derivatives thereof on the same polynucleotide molecule or on separate polynucleotide molecules. Additionally, the polynucleotides of the present disclosuremayencode portions of the heavy and light chain variable regions of the antigen-binding polypeptides, variants or derivatives thereof on the same polynucleotide molecule or on separate polynucleotide molecules.

[0093] Methods of making antibodies are well known in the art and described herein. In certain embodiments, both the variable and constant regions of the antigen-binding polypeptides of the present disclosure are fully human. Fully human antibodies can be made using techniques described in the art and as described herein. For example, fully human antibodies against a specific antigen can be prepared by administering the antigen to a transgenic animal which has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled. Exemplary techniques that can be used to make such antibodies are described in U.S. patents: 6,150,584; 6,458,592; 6,420,140 which are incorporated by reference in their entireties. Treatment Methods

[0094] As described herein, the antibodies, variants or derivatives of the present disclosure may be used in certain treatment and diagnostic methods.

[0095] Thepresentdisclosure is further directed toantibody-based therapieswhich involveadministering theantibodies of the disclosure to a patient such as an animal, a mammal, and a human for treating one or more of the disorders or conditions described herein. Therapeutic compounds of the disclosure include, but are not limited to, antibodies of the disclosure (including variants and derivatives thereof as described herein) and nucleic acids or polynucleotides encoding antibodies of the disclosure (including variants and derivatives thereof as described herein).

[0096] The antibodies of the disclosure can also be used to treat or inhibit cancer. As provided above, GPRC5D can be overexpressed in cancer cells, in particular multiple myeloma. Inhibition of GPRC5D has been shown to be useful for treating the cancers.

[0097] Accordingly, in some embodiments, provided aremethods for treating a cancer in a patient in need thereof. The method, in one embodiment, entails administering to the patient an effective amount of an antibody of the present disclosure. In some embodiments, at least one of the cancer cells in the patient over-express GPRC5D. In some embodiments, the antibody or fragment is ADCC-competent. In some embodiments, the antibody or fragment further comprisesacytotoxic agent. In someembodiments, theantibody isbispecific that further targetsan immunecell, suchasa cytotoxic T cell. 13 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55

[0098] Cellular therapies, such as chimeric antigen receptor (CAR) T-cell ( or NK cell, macrophage) therapies, are also provided in the present disclosure. A suitable cell can be used, that is put in contact with an anti-GPRC5D antibody of the present disclosure (or alternatively engineered to express an anti-GPRC5D antibody of the present disclosure). Upon such contact or engineering, the cell can then be introduced to a cancer patient in need of a treatment. The cancer patient may have a cancer of any of the types as disclosed herein. The cell (e.g., Tcell) can be, for instance, a tumor-infiltrating T lymphocyte, a CD4+ T cell, a CD8+ T cell, a natural killer (NK) cell, a macrophage, or the combination thereof, without limitation.

[0099] In some embodiments, the cell was isolated from the cancer patient him‑ or her-self. In some embodiments, the cell was provided by a donor or from a cell bank. When the cell is isolated from the cancer patient, undesired immune reactions can be minimized.

[0100] Non-limiting examples of cancers include hematological cancers such as multiple myeloma. Other examples include leukemia (including acute leukemias (e.g., acute lymphocytic leukemia, acute myelocytic leukemia (including myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroleukemia)) and chronic leukemias (e.g., chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia)), and lymphomas (e.g., Hodgkin’s disease and non-Hodgkin’s disease), and multiple myeloma.

[0101] Aspecific dosageand treatment regimen for any particular patientwill depend upona variety of factors, including the particular antibodies, variant or derivative thereof used, the patient’s age, body weight, general health, sex, and diet, and the time of administration, rate of excretion, drug combination, and the severity of the particular disease being treated. Judgment of such factors by medical caregivers is within the ordinary skill in the art. The amount will also depend on the individual patient to be treated, the route of administration, the type of formulation, the characteristics of the compound used, the severity of the disease, and the desired effect. The amount used can be determined by pharmacological and pharmacokinetic principles well known in the art.

[0102] Methods of administration of the antibodies, variants or include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The antigen-binding polypeptides or compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be adminis- tered together with other biologically active agents. Thus, pharmaceutical compositions containing the antigen-binding polypeptides of the disclosure may be administered orally, rectally, parenterally, intracistemally, intravaginally, intraper- itoneally, topically (as by powders, ointments, drops or transdermal patch), bucally, or as an oral or nasal spray.

[0103] The term "parenteral" asusedherein refers tomodesof administrationwhich include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intra-articular injection and infusion.

[0104] Administration can be systemic or local. In addition, it may be desirable to introduce the antibodies of the disclosure into the central nervous system by any suitable route, including intraventricular and intrathecal injection; intraventricular injectionmay be facilitated by an intraventricular catheter, for example, attached to a reservoir, such as an Ommaya reservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.

[0105] Itmay bedesirable to administer the antigen-binding polypeptides or compositions of the disclosure locally to the area in need of treatment; thismay be achieved by, for example, and not byway of limitation, local infusion during surgery, topical application,e.g., in conjunction,with awounddressingafter surgery, by injection, bymeansof a catheter, bymeans of a suppository, or by means of an implant, said implant being of a porous, nonporous, or gelatinous material, including membranes, suchassialasticmembranes, or fibers.Preferably,whenadministeringaprotein, includinganantibody, of the disclosure, care must be taken to use materials to which the protein does not absorb.

[0106] Theamount of the antibodies of the disclosurewhichwill be effective in the treatment, inhibition andprevention of an inflammatory, immuneormalignant disease,disorderor conditioncanbedeterminedbystandardclinical techniques. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulationwill also dependon the route of administration, and the seriousnessof the disease, disorder or condition, and should bedecided according to the judgment of the practitioner andeachpatient’s circumstances. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0107] Asageneral proposition, the dosage administered to a patient of the antigen-binding polypeptides of the present disclosure is typically 0.1mg / kg to 100mg / kgof thepatient’sbodyweight, between0.1mg / kgand20mg / kgof thepatient’s bodyweight, or 1mg / kg to 10mg / kgof the patient’sbodyweight.Generally, humanantibodieshavea longer half-lifewithin the human body than antibodies from other species due to the immune response to the foreign polypeptides. Thus, lower dosages of human antibodies and less frequent administration is often possible. Further, the dosage and frequency of administration of antibodies of the disclosure may be reduced by enhancing uptake and tissue penetration (e.g., into the brain) of the antibodies by modifications such as, for example, lipidation.

[0108] In an additional embodiment, the compositions of the disclosure are administered in combinationwith cytokines. Cytokines that may be administered with the compositions of the disclosure include, but are not limited to, IL‑2, IL‑3, IL‑4, IL‑5, IL‑6, IL‑7, IL‑10, IL‑12, IL‑13, IL‑15, anti-CD40, CD40L, and TNF-α. 14 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55

[0109] In additional embodiments, the compositions of the disclosure are administered in combination with other therapeutic or prophylactic regimens, such as, for example, radiation therapy. Diagnostic Methods

[0110] Over-expression of GPRC5D is observed in certain tumor samples, and patients having GPRC5D-over- expressing cells are likely responsive to treatments with the anti-GPRC5D antibodies of the present disclosure. Accordingly, the antibodies of the present disclosure can also be used for diagnostic and prognostic purposes.

[0111] A sample that preferably includes a cell can be obtained froma patient, which can be a cancer patient or a patient desiring diagnosis. The cell be a cell of a tumor tissue or a tumor block, a blood sample, a urine sample or any sample from the patient. Upon optional pretreatment of the sample, the sample can be incubated with an antibody of the present disclosure under conditions allowing the antibody to interact with a GPRC5D protein potentially present in the sample. Methods such as ELISA can be used, taking advantage of the anti-GPRC5D antibody, to detect the presence of the GPRC5D protein in the sample.

[0112] Presence of the GPRC5D protein in the sample (optionally with the amount or concentration) can be used for diagnosis of cancer, as an indication that the patient is suitable for a treatmentwith the antibody, or as an indication that the patient has (or has not) responded to a cancer treatment. For a prognostic method, the detection can be done at once, twice or more, at certain stages, upon initiation of a cancer treatment to indicate the progress of the treatment. Compositions

[0113] The present disclosure also provides pharmaceutical compositions. Such compositions comprise an effective amount of an antibody, and an acceptable carrier. In some embodiments, the composition further includes a second anticancer agent (e.g., an immune checkpoint inhibitor).

[0114] In a specific embodiment, the term "pharmaceutically acceptable"meansapprovedbya regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. Further, a "pharmaceutically acceptable carrier" will generally be a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0115] The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Suchpharmaceutical carriers canbesterile liquids, suchaswaterandoils, including thoseof petroleum,animal, vegetable or synthetic origin, suchaspeanut oil, soybeanoil,mineral oil, sesameoil and the like.Water is apreferred carrierwhen the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skimmilk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates.Antibacterial agents suchasbenzyl alcohol ormethyl parabens; antioxidants suchasascorbic acidor sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and agents for the adjustment of tonicity such as sodium chloride or dextrose are also envisioned. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences by E. W. Martin, incorporated herein by reference. Such compositions will contain a ther- apeutically effectiveamount of theantigen-bindingpolypeptide, preferably inpurified form, togetherwithasuitableamount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration. The parental preparation canbeenclosed in ampoules, disposable syringes ormultiple dose vialsmadeof glass or plastic.

[0116] In an embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous adminis- tration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline.Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

[0117] The compounds of the disclosure can be formulated as neutral or salt forms. Pharmaceutically acceptable salts 15 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc. EXAMPLES Example 1: Generation of murine monoclonal antibodies against human GPRC5D

[0118] The human GPRC5D protein was used to immunize different strains of mice and hybridomas were generated accordingly. More than twenty hybridoma clones were collected for further analysis.

[0119] Antibodies harvested from the supernatants of the hybridomas were tested for their binding to the human GPRC5D protein expressed on CHO K1 cells was tested. CHO-K1 cells stably expressed human GPRC5D, were harvested from flasks. 100 µl of 1×106 cells / ml of cells were incubated with murine antibodies in 3-fold serial dilutions for 30minutes on ice. After being washed with 200µl of FACS buffer twice, cells were incubated with secondary antibody for 30minuteson ice.Cellswerewashedwith200µl of FACSbuffer twiceand transferred toBDFalcon5ml tubeandanalyzed byFACS. The results of the study showed that themurine antibodies can bind to humanGPRC5Dwith highEC50 (FIG.1 ). The results are shown in the table below. All of them exhibited good binding activity. Bottom Top LogEC50 HillSlope EC50 Span 2D3C12 46110 835137 0.4535 1.132 2.841 789027 6G10D9 6870 1429638 0.9116 0.9535 8.159 1422768 11C1H10 40235 859162 0.5274 1.101 3.368 818927 14C2A11C6 5573 728662 0.9934 0.8801 9.849 723089 14C2F7C6 15616 751279 0.9634 0.9615 9.193 735663 14F3H2 20428 872785 0.8744 1.146 7.489 852357 21C3C11B3 57013 1508205 0.6538 1.1 4.506 1451192 26E1D12 52742 677713 0.2246 1.303 1.677 624970 29A7H11B9D3 71394 1385928 0.4219 1.259 2.642 1314534 29A7H11H4C12 43439 1461931 0.7377 1.109 5.466 1418492 29B2C10 68634 1607305 0.6201 1.305 4.17 1538671 31H11C10H7 16791 961550 0.9534 1.134 8.983 944759 34D3F8B5 ‑16109 1407656 1.084 0.7475 12.12 1423765 34D3H1 ‑13314 1412131 0.8843 0.8042 7.662 1425446 34D4A10 2737 921851 1.019 0.8496 10.44 919114 36F11E9H3 13852 766682 0.8732 1.087 7.468 752830 37B9C4 10880 1758861 0.5917 0.9933 3.906 1747981 38A10E9A11 24016 1785041 0.9472 1.099 8.854 1761025 39D2B6 7833 1774093 0.5469 0.98 3.523 1766260 40A5B2 25394 1196144 0.5108 0.9236 3.242 1170750 57G11D4A10 ‑22363 2089505 0.9703 0.8141 9.34 2111868 58F9G10 ‑14744 2052463 0.9635 0.8313 9.194 2067207

[0120] Fourteen hybridomas were subcloned and the VH / VL sequences were determined (see Table 1). 16 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 Table 1. VH / VL sequence of the lead murine antibodies Name Sequence (CDRs are underlined) SEQ ID NO: 6G10D9 VH 1 6G10D9 VL 2 21C3C11B3 VH 3 21C3C11B3 VL 4 29B2C10 VH 5 29B2C10 VL 6 34D3H1 VH 7 34D3H1 VL 8 37B9C4 VH 9 37B9C4 VL 10 38A10E9A11 VH 11 38A10E9A11 VL 12 40A5B2 VH 13 40A5B2 VL 14 58F9G10 VH 15 17 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Name Sequence (CDRs are underlined) SEQ ID NO: 58F9G10 VL 16 2D3C12 VH 17 2D3C12 VL 18 14C2A11C6 VH 19 14C2A11C6 VL 20 14F3H2 VH 21 14F3H2 VL 22 26E1D12 VH 23 26E1D12 VL 24 29A7H11H4C12 VH 25 29A7H11H4C12 VL 26 34D4A10 VH 27 34D4A10 VL 28 Example 2. Binding of Chimeric Antibodies to GPRC5D

[0121] Themurine VH and VK genes were produced synthetically and then respectively cloned into vectors containing the human gamma 1 and human kappa constant domains. The purified chimeric antibodies were produced from transfected CHOs cells.

[0122] CHO-K1 cells stably expressed human GPRC5D, were harvested from flasks. 100 µl of 1×106 cells / ml of cells were incubated with primary chimeric antibodies in 3-fold serial dilutions starting from 300 nM to 0.001 nM for 30 minutes on ice. After being washed with 200 µl of FACS buffer twice, cells were incubated with secondary antibody for 30minutes 18 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 on ice. Cells werewashedwith 200µl of FACSbuffer twice and transferred to BDFalcon 5ml tube and analyzed by FACS. The results of the study showed that the chimeric antibodies can bind to human GPRC5D with high EC50 (FIG. 2 ).

[0123] The results are shown in the table below. Bottom Top LogEC50 HillSlope EC50 Span 58F9G10 ‑521.8 414579 0.3596 0.8258 2.289 415101 40A5B2 1487 440269 0.3436 0.8189 2.206 438781 38A9E10A11 3016 386984 0.2453 1.037 1.759 383968 37B9C4 2078 405062 0.249 1.045 1.774 402985 34D3H1 3448 507844 0.5214 1.012 3.322 504396 29B2C10 4326 430443 0.342 1.041 2.198 426116 21C3C11B3 5774 403938 0.2293 1.286 1.696 398164 6G10D9 6886 458124 0.1012 1.343 1.262 451238 Example 3. EC50 of internalization of chimeric GPRC5D antibodies on CHO-GPRCSD cells

[0124] pHAbDyesarepHsensordyes that havevery lowfluorescenceat pH>7andadramatic increase in fluorescence as thepHof the solution becomesacidic. pHAbDyeshaveexcitationmaxima (Ex) at 532nmandemissionmaxima (Em)at 560nm. pHAb Dye-conjugated antibodies can be used for monitoring receptor-mediated antibody internalization. When an antibody-pHAb Dye conjugate binds to its receptor on the cell membrane, it exhibits minimal fluorescence. However, upon receptor-mediated internalization, antibody-pHAb Dye conjugates traffic to the endosome and lysosomal vesicles where pH is acidic, causing the pHAb Dye to fluoresce. This fluorescence can be detected using various techniques, including cell imaging, flow cytometry and fluorescent plate-based readers with appropriate filters.

[0125] Stably transfected human GPRC5D CHO cells were harvested with 0.05% Trypsin / EDTA (Gibco, 25300‑054) and plated in a 96-well black plate (Thermo Scientific #165305) at the density of 10 K per 90 µl per well. Plates were incubated for 20‑24 h before treatment with pHAb labeled antibodies.

[0126] For internalization, pHAbconjugated chimericGPRC5Dantibodieswereadded to thecells at different andmixed gently for 1‑2minonaplatemixer and then incubatedovernight to allow internalization (internalization canbedetected in a few hours). Plates were read on a fluorescent plate reader at Ex / Em: 532 nm / 560 nm on a Tecan Infinity M1000 Pro. To achieve higher sensitivity, media was replaced by PBS before reading the plate.

[0127] The results normalizedwithDARareshown in theFIG. 3.Asshown inFIG. 3, the testedchimeric antibodieshave potent internalization activity. Example 4. Testing of Antibody-Dependent Cellular Cytotoxicity (ADCC)

[0128] The ADCC Reporter Bioassay uses an alternative readout at an earlier point in ADCC MOA pathway activa- tion:theactivationof gene transcription through theNFAT (nuclear factor of activatedT-cells) pathway in theeffector cell. In addition, the ADCC Reporter Bioassay uses engineered Jurkat cells stably expressing the FcγRIIIa receptor, V158 (high affinity) variant, andanNFATresponseelement driving expression of firefly luciferase as effector cells. Antibodybiological activity in ADCC MOA is quantified through the luciferase produced as a result of NFAT pathway activation; luciferase activity in the effector cell is quantified with luminescence readout. Signal is high, and assay background is low.

[0129] Serial dilutions of GPRC5D chimeric monoclonal antibody were incubated for 6 hours of induction at 37°C with engineered Jurkat effector cells (ADCCBioassayEffector Cells), with or without ADCCBioassay Target Cells (GPRC5D). Luciferase activity was quantified using Bio-Glo™ Reagent (in FIG. 4). All of the tested antibodies exhibited potent capability in inducing ADCC. Example 5. Humanization of the mouse mAbs

[0130] The murine antibody variable region genes were employed to create humanized mAbs. In the first step of this process, the amino acid sequences of the VH and VL of mAb were compared against the available database of human Ig gene sequences to find the overall best-matching human germline Ig gene sequences.

[0131] The amino acid sequences of the humanized antibody are provided below. Humanized sequences 19 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 A. 34D3H1

[0132] Table 2A. Humanization of 34D3H1 - VH Name Sequence SEQ ID NO: 34D3H1 VH 7 V1 (CDR grafting) 35 V2 (with back mutations) 36 V3 (with back mutations) 37 Table 2B. CDR Sequences CDR Sequence SEQ ID NO: CDR-H1 SYGMS 29 CDR-H2 TISSGGSYTYYPDSVKG 30 CDR-H3 QGGDAMDY 31 Table 2C. Humanization of 34D3H1 - VL Name Sequence SEQ ID NO: 34D3H1 VL 8 V1 (CDR grafting) 38 V2 (with back mutations) 39 V3 (with back mutations) 40 V4 (with back mutations) 41 Table 2D. CDR Sequences CDR Sequence SEQ ID NO: CDR-L1 RASQSINNNLH 32 CDR-L2 YASQSIS 33 20 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) CDR Sequence SEQ ID NO: CDR-L3 QQSNSRLT 34 Table 2E. Humanized antibodies VL VL v1 VL v2 VL v3 VL v4 VH 34-XI VH v1 34-H1L1 34-HIL2 34-H1L3 34-H1L4 VH v2 34-H2L1 34-H2L2 34-H2L3 34-H2L4 VH v3 34-H3L1 34-H3L2 34-H3L3 34-H3L4 B. 37B9C4

[0133] Table 3A. Humanization of 37B9C4 - VH Name Sequence SEQ ID NO: 37B9C4 VH 9 V1 (CDR grafting) 48 V2 (with back mutations) 49 V3 (with back mutations) 50 Table 3B. CDR Sequences CDR Sequence SEQ ID NO: CDR-H1 DYWMN 42 CDR-H2 EIRLKSNNYATHYAESVKG 43 CDR-H3 PLLWFRRYYAMDY 44 Table 3C. Humanization of 37B9C4 - VL Name Sequence SEQ ID NO: 37B9C4 VL 10 V1 (CDR grafting) 51 21 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Name Sequence SEQ ID NO: V2 (with back mutations) 52 V3 (with back mutations) 53 Table 3D. CDR Sequences CDR Sequence SEQ ID NO: CDR-L1 SASQGISNYLN 45 CDR-L2 YTSSLHS 46 CDR-L3 QQYSKLPFT 47 Table 3E. Humanized antibodies VL VL v1 VL v2 VL v3 VH 37B9C4-XI VH v1 37-H1L1 37-H1L2 37-H1L3 VH v2 37-H2L1 37-H2L2 37-H2L3 VH v3 37-H3L1 37-H3L2 37-H3L3 C. 58F9G10

[0134] Table 4A. Humanization of 58F9G10 - VH Name Sequence SEQ ID NO: 58F9G10 VH 15 V1 (CDR grafting) 61 V2 (with back mutations) 62 V3 (with back mutations) 63 V4 (with back mutations) 64 22 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 Table 4B. CDR Sequences CDR Sequence SEQ ID NO: CDR-H1 GYYIH 54 CDR-H2 YISCYNGATSFNQKFKG 60 YISSYNAATSFNQKFKG 55 CDR-H3 TELRGPWFAY 56 Table 4C. Humanization of 58F9G10 - VL Name Sequence SEQ ID NO: 58F9G10 VL 16 V1 (CDR grafting) 65 V2 (with back mutations) 66 V3 (with back mutations) 67 Table 4D. CDR Sequences CDR Sequence SEQ ID NO: CDR-L1 SASSSVSYMN 57 CDR-L2 DTSKLAS 58 CDR-L3 QQWSNNPLT 59 Table 3E. Humanized antibodies VL VL v1 VL v2 VL V3 VH 58F9G10-XI VH v1 58-H1L1 58-H1L2 58-H1L3 VH v2 58-H2L1 58-H2L2 58-H2L3 VH v3 58-H3L1 58-H3L2 58-H3L3 VH v4 58-H4L1 58-H4L2 58-H4L3 D. 6G10D9

[0135] Table 5A. Humanization of 6G10D9 - VH Name Sequence SEQ ID NO: 6G10D9 VH 1 23 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Name Sequence SEQ ID NO: V1 (CDR grafting) 74 V2 (with back mutations) 75 V3 (with back mutations) 76 V4 (with back mutations) 77 V5 (with back mutations) 78 V6 (with back mutations) 79 Table 5B. CDR Sequences CDR Sequence SEQ ID NO: CDR-H1 TYTMH D01 CDR-H2 YINPSSGYTNYNQKFKD D02 CDR-H3 LRSRGYFDY D03 Table 5C. Humanization of 6G10D9 - VL Name Sequence SEQ ID NO: 6G10D9 VL 2 V1 (CDR grafting) 80 V2 (with back mutations) 81 V3 (with back mutations) 82 V4 (with back mutations) 83 V1a (CDR grafting version a) 84 24 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Name Sequence SEQ ID NO: V2a (with back mutations) 85 V3a (with back mutations) 86 Table 5D. CDR Sequences CDR Sequence SEQ ID NO: CDR-L1 KASONVGTAVV 71 CDR-L2 SASNRYT 72 CDR-L3 QQYSSYPYT 73 Table 5E. Humanized antibodies VL VL v1 VL v2 VL v3 VL v4 VL v1a VL v2a VL v3a VH 6G10D9-XI VH v1 6-H1L1 6-H1L2 6-H1L3 VH v2 6-H2L1 6-H2L2 6-H2L3 VH v3 6-H3L1 6-H3L2 6-H3L3 VH v4 6-H4L1 6-H4L2 6-H4L3 VH v5 VH v6 Example 6. Testing of Humanized Antibodies

[0136] This example tested some of the humanized antibodies for the ability to bind toGPRC5D expressed onCHO-K1 cells.

[0137] Out of the tested humanized antibodies derived from6G10D9, 6-H2L1 and 6-H3L3 outperformed others (FIG. 5, Table 6). Table 6. Activities of humanized antibodies of 6G10D9 to bind GPRC5D Bottom Top LogEC50 HillSlope EC50 Span 6G10D9-XI 6858 121725 0.2992 1.525 1.991 114867 6-H1L1 192.6 ~ 6397 ~ 3.102 1.198 ~ 1264 ~ 6205 6-H1L2 191.9 898894 5.224 1.19 167639 898702 6-H1L3 226.2 16478302 5.389 1.156 244790 16478076 6-H2L1 189.1 661560 5.039 1.232 109294 661371 6-H2L2 192.5 1271084 5.37 1.191 234364 1270892 6-H2L3 215.7 10592380 5.039 1.23 109279 10592164 6-H3L1 ‑205.1 74293 2.352 0.8392 225.1 74498 6-H3L2 68.94 47463 2.244 0.9532 175.3 47394 6-H3L3 3994 119304 0.9553 1.13 9.022 115310 6-H4L1 61.5 50293 1.654 0.9277 45.05 50231 25 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Bottom Top LogEC50 HillSlope EC50 Span 6-H4L2 9.56 39449 1.976 0.911 94.63 39440 6-H4L3 2344 126413 0.9049 1.052 8.033 124069

[0138] As shown inFIG. 6 andTable 7, all of the humanized versions of 58F9G10 appeared to have good performance. Table 7. Activities of humanized antibodies of 58F9G10 to bind GPRC5D Bottom Top LogEC50 HillSlope EC50 Span 58F9G10-XI 6697 127349 0.2925 1.292 1.961 120652 58-H1L1 15047 107881 0.2519 1.342 1.786 92834 58-H1L2 5500 114432 0.2632 1.238 1.833 108932 58-H1L3 4227 105346 0.2308 1.14 1.701 101119 58-H2L1 3617 109672 0.3776 1.064 2.386 106055 58-H2L2 4837 107888 0.3448 1.091 2.212 103050 58-H2L3 1229 91713 0.2697 0.9633 1.861 90483 58-H3L1 9887 103663 ‑0.1918 1.382 0.643 93776 58-H3L2 5981 105458 0.3742 1.195 2.367 99476 58-H3L3 4022 97718 0.3177 1.105 2.078 93695 58-H4L1 4993 104559 0.3655 1.179 2.32 99566 58-H4L2 4600 105051 0.3407 1.143 2.191 100451 58-H4L3 6249 102650 0.2874 1.27 1.938 96401

[0139] Likewise, as shown in FIG. 7 and Table 8, all of the humanized versions of 34D3H1 appeared to have good performance, comparable to the chimeric antibody. Table 8. Activities of humanized antibodies of 34D3H1 to bind GPRC5D Bottom Top LogEC50 HillSlope EC50 Span 34-XI 16525 192054 0.4947 1.454 3.124 175530 34-H1L1 10921 196570 0.5908 1.314 3.898 185650 34-H1L2 13856 182030 0.5836 1.436 3.834 168174 34-H1L3 10620 185840 0.5732 1.167 3.743 175220 34-H1L4 10299 188925 0.5999 1.195 3.98 178626 34-H2L1 7854 187153 0.6188 1.173 4.157 179298 34-H2L2 8736 178873 0.6254 1.118 4.221 170137 34-H2L3 7063 185842 0.5832 1.11 3.83 178779 34-H2L4 4393 192405 0.6092 1.041 4.066 188012 34-H3L1 11139 178248 0.5207 1.228 3.317 167109 34-H3L2 9036 179809 0.6464 1.146 4.43 170773 34-H3L3 9118 192976 0.6508 1.1 4.475 183858 34-H3L4 12751 192533 0.5981 1.263 3.963 179782

[0140] Also, as shown in FIG. 8 and Table 9, all of the humanized versions of 37B9C4 appeared to have good 26 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 performance, comparable to the chimeric counterpart. Table 9. Activities of humanized antibodies of 37B9C4 to bind GPRC5D Bottom Top LogEC50 HillSlope EC50 Span 37B9C4-XI 11711 188479 0.2364 1.502 1.723 176769 37-H1L1 9596 178413 0.3574 1.613 2.277 168817 37-H1L2 8848 181865 0.3502 1.534 2.24 173017 37-H1L3 10445 173229 0.2332 1.528 1.711 162784 37-H2L1 9184 175072 0.351 1.583 2.244 165888 37-H2L2 9383 175293 0.3455 1.656 2.216 165910 37-H2L3 8049 168032 0.3334 1.564 2.155 159982 37-H3L1 6372 179261 0.2921 1.35 1.96 172889 37-H3L2 9193 179304 0.3153 1.428 2.067 170111 37-H3L3 7780 181053 0.2846 1.305 1.926 173273

[0141] Based on the above data, humanized antibodies 6-H3L3, 6-H4L3, 58-H1L1, 58-H3L1, 34-H1L1 and 37-H1L1 were selected for further confirmative testing. Example 7. Confirmative Testing of Select Humanized Antibodies

[0142] This example tested some of the humanized antibodies (6-H3L3, 6-H4L3, 58-H1L1, 58-H3L1, 34-H1L1 and 37- H1L1) for their ability to bind to the human GPRC5D protein expressed on CHO-K1 and NCI-H929 cells.

[0143] The binding data to CHO-K1 are charted in FIG. 9 and summarized in Table 10 below. Table 10. Binding to GPRC5D on CHO-K1 cells Bottom Top LogEC50 HillSlope EC50 Span 6-XI 523.9 104077 0.4723 1.095 2.967 103553 58-XI 52.25 107000 0.5562 0.9481 3.599 106948 6-H3L3 172 134045 1.412 0.8045 25.82 133873 6-H4L3 109.4 145833 1.421 0.7392 26.34 145724 58-H1L1 275.4 99454 0.579 0.972 3.793 99178 58-H3L1 75.37 104402 0.612 0.9135 4.093 104326 34-XI 688.6 117714 0.5776 1.054 3.781 117025 37-XI 792.6 113379 0.3877 1.175 2.442 112586 34-H1L1 170.8 119038 0.781 0.8745 6.04 118867 37-H1L1 523.2 102496 0.511 1.042 3.244 101973 Table 11. Binding to GPRC5D on NCI-H929 cells Bottom Top LogEC50 HillSlope EC50 Span 6-XI 269.4 13785 0.05146 1.313 1.126 13515 58-XI 261.4 14738 0.1596 1.269 1.444 14476 6-H3L3 129.8 21859 1.465 0.8063 29.18 21730 6-H4L3 80.05 28987 1.606 0.7142 40.33 28907 58-H1L1 281 13474 0.1648 1.319 1.462 13193 58-H3L1 275.5 13934 0.3 1.297 1.995 13658 27 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Bottom Top LogEC50 HillSlope EC50 Span 34-XI 181.1 18146 0.7587 0.9911 5.738 17965 34-H1L1 199.3 17911 0.8048 1.011 6.379 17712 37-XI 337.6 13807 ‑0.05621 1.41 0.8786 13470 37-H1L1 341.8 13534 ‑0.1991 1.419 0.6323 13192

[0144] These data, therefore, confirm that these selected humanized antibodies are suitable for further clinical development. Example 8. ADCC of Humanized Antibodies

[0145] The ADCCReporter Bioassay uses an alternative readout at an earlier point in ADCCMOA pathway activation: the activation of gene transcription through the NFAT (nuclear factor of activated T-cells) pathway in the effector cell. In addition, the ADCC Reporter Bioassay uses engineered Jurkat cells stably expressing the FcγRIIIa receptor, V158 (high affinity) variant, andanNFATresponseelement driving expression of firefly luciferase as effector cells. Antibodybiological activity in ADCC MOA is quantified through the luciferase produced as a result of NFAT pathway activation; luciferase activity in the effector cell is quantified with luminescence readout. Signal is high, and assay background is low.

[0146] Serial dilutions of GPRC5D hu Abs were incubated for 6 hours of induction at 37°C with engineered Jurkat effector cells (ADCC Bioassay Effector Cells), with ADCC Bioassay Target Cells (expressing GPRC5D). Luciferase activity was quantified using Bio-GloTM Reagent.

[0147] The results are presented in FIG. 11, which shows that these humanized antibodies induced strong ADCC activities on GPRC5D-overexpressing cell lines and endogenous MM (multiple myeloma) cell lines. Example 9. Internalization of Humanized Antibodies

[0148] This example tested the humanized antibodies for their internationalization induction activities. The example used a new hydrophilic and bright pH sensor dye (pHAb dye), which is not fluorescent at neutral pH but becomes highly fluorescent at acidic pH with internalization. It can be used to detect the process of internalization. NCI-H929 andMM.1R cells endogenously expressed human GPRC5D as the target cells, the detection antibody labeled with pHAb dye was added to evaluate the internalization of GPRC5D hu antibody in vitro.

[0149] Serial dilutions of GPRC5D hu antibodieswere incubated for 24 hours at 37°C. Luciferase activity was detected. The results, inFIG. 12, show that these humanized antibodies have very strong internalization activities to bothGPRC5D- overexpressing cell lines and endogenous MM cell lines. Example 10. Killing Activity of GPRC5D ADC

[0150] This example tested two antibody-drug conjugates (ADC) for their killing activities of targeted cells. These ADC included 37B9C4 and 58F9G10, respectively, conjugated to the toxic drug Monomethyl auristatin E (MMAE).

[0151] The human GPRC5D expressing HEK293 and NCI-H929 engineering cells (HEK293 / H_GPRC5D and NCI- H929 / H_GPRC5D), GPRC5D endogenously expressing NCI-H929 and MM.1R cells were seeded to a 96-well plate at 3000~4000 cells per well. The cells were treated with 37B9C4-MMAE and 58F9G10-MMAE at respective concentrations for 5 days. The cell viability was measured by CellTiteseeded to a 96-well plate at 3000~4000 cells per well with r-Glo reagent. The luciferase activitywasdetectedbyEnvison. The results, inFIG. 13andTable 12, show that thesehumanized antibodies have very strong killing activities. Table 12. ADC Killing Activities Hek‑293 / H_GPRC5D (IC50, nM) MM.1R (IC50, nM) NCI-H929 (IC50, nM) NCI-H929 / GPRCSD (IC50, nM) 37B9C4-MMAE 0.025 0.14 0.23 0.11 58F9G10-MMAE 0.026 0.07 0.99 0.14 28 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 Example 11. In vivo Anti-Tumor Activities

[0152] This example used a CDX animal model to test the anti-tumor activities of antibody 37B9C4 (H1L1).

[0153] In this study, 6‑8 weeks female NCG mice (Jiangsu Jicui Yaokang Biotechnology Co., Ltd) were used. Each mouse was inoculated subcutaneously at the right axillary (lateral) with MM.1R tumor cell (2×106) in 0.1 ml of PBS with matrigel (V:V=1:1) for tumor development. The animals were randomly grouped when tumor volume reach around 60 mm3, then treatment started for the efficacy study. 37B9C4 at dose of 1mg / kg, 3mg / kg and 10 mg / kg were administrated via intravenous (i.v.) at day 0, day 7, day 14. The experiment was terminated at day 19 when the average tumor volume in vehicle groupwas over 2000mm3. The average tumor volumeof PBSgroup, 37B9C4 (1mg / kg) group, 37B9C4 (3mg / kg) group and 37B9C4 (10 mg / kg) group were 2498.58 mm3, 1196.15 mm3, 0.00 mm3 (6 / 6 CR), and 0.00 mm3 (6 / 6 CR) respectively. The tumor size were measured three times weekly in two dimensions using a caliper and the volume was expressed in mm3 using the formula: V = 0.5 a x b2 where a and b were the long and short diameters of the tumor, respectively. Data points represent group (n=6) mean, error bars represent standard error of the mean (SEM).

[0154] The body weight of MM.1R tumor bearing mice was monitored regularly as an indirect measure of toxicity. FIG. 14A shows the tumor growth curve of MM.1R tumor-bearing mice post administration of 37B9C4. The antibody dose- dependently inhibited the tumor growth in this tumor model, and achieved complete tumor inhibition at or above 3mg / kg.

[0155] The detailed changes of body weight and relative change of body weight of MM.1R tumor bearing mice after administration are shown in FIG. 14B.During the administration period, all groups of mice had no significant body weight loss, and the mice in the administration group had good tolerance, demonstrating the safety of the treatment. CLAUSES

[0156] An antibody or antigen-binding fragment thereof having binding specificity to a human G-protein-coupled receptor family C group 5 member D (GPRC5D) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region (VH) comprising heavy chain complementarity determining regions CDRH1, CDRH2, and CDRH3 and a light chain variable region (VL) comprising complementarity determining regionsCDRL1,CDRL2, andCDRL3, and wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise: (a) the amino acid sequences of SEQ ID NO:42‑47; (b) the amino acid sequences of SEQ ID NO:29‑34; (c) the amino acid sequences of SEQ ID NO:54‑59; or (d) the amino acid sequences of SEQ ID NO:68‑73. 2. The antibody or antigen-binding fragment thereof of clause 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:42‑47. 3. The antibody or antigen-binding fragment thereof of clause 2, wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:9 and 48‑50, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:10 and 51‑53. 4. The antibody or antigen-binding fragment thereof of clause 2, wherein the VH comprises the amino acid sequence of SEQ ID NO:48, and the VL comprises the amino acid sequence of SEQ ID NO:51. 5. The antibody or antigen-binding fragment thereof of clause 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:29‑34. 6. The antibody or antigen-binding fragment thereof of clause 5, wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:7 and 35‑37, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:8 and 38‑41. 7. The antibody or antigen-binding fragment thereof of clause 5, wherein the VH comprises the amino acid sequence of SEQ ID NO:35, and the VL comprises the amino acid sequence of SEQ ID NO:38. 8. The antibody or antigen-binding fragment thereof of clause 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:54‑59. 9. The antibody or antigen-binding fragment thereof of clause 8, wherein the VH comprises an amino acid sequence selected from thegroup consisting of SEQ IDNO:61‑64, and theVLcomprises anaminoacid sequenceselected from 29 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 the group consisting of SEQ ID NO:16 and 65‑67. 10. Theantibody or antigen-binding fragment thereof of clause8,wherein theVHcomprises theaminoacid sequence of SEQ ID NO:61, and the VL comprises the amino acid sequence of SEQ ID NO:65. 11. The antibody or antigen-binding fragment thereof of clause 8,wherein theVHcomprises the aminoacid sequence of SEQ ID NO:63, and the VL comprises the amino acid sequence of SEQ ID NO:65. 12. The antibody or antigen-binding fragment thereof of clause 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:68‑73. 13.Theantibodyorantigen-binding fragment thereof of clause12,wherein theVHcomprisesanaminoacidsequence selected from the group consisting of SEQ ID NO:1 and 74‑79, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2 and 80‑86. 14. The antibody or antigen-binding fragment thereof of clause 12, wherein the VH comprises the amino acid sequence of SEQ ID NO:76, and the VL comprises the amino acid sequence of SEQ ID NO:82. 15. The antibody or antigen-binding fragment thereof of clause 12, wherein the VH comprises the amino acid sequence of SEQ ID NO:77, and the VL comprises the amino acid sequence of SEQ ID NO:82. 16. The antibody or fragment thereof of any one of clauses 1‑15, which is humanized. 17. The antibody or fragment thereof of any one of clauses 1‑16, which is ADCC-competent. 18. The antibody or fragment thereof of any one of clauses 1‑16, further comprising a cytotoxic drug conjugated to the antibody or fragment thereof. 19. A bispecific antibody, comprising the antigen-binding fragment of any one of clauses 1‑16, and a second antigen- binding fragment having specificity to a second target protein. 20. The bispecific antibody of clause 19, wherein the second target protein is selected from the group consisting of CD3, CD16, CD19, CD28, CD64 and 4‑1BB. 21. The bispecific antibody of clause 19, wherein the second target protein is CD3. 22. A method of treating cancer in a patient in need thereof, comprising administering to the patient the antibody or fragment thereof of any one of clauses 1‑21. 23.Amethodof treatingcancer in apatient in need thereof, comprising (a) treatingaTcell, a natural killer (NK) cell, or a macrophage, in vitro, with the antibody or fragment thereof of any one of clauses 1‑21, and (b) administering the treated cell to the patient. 24. Themethod of clause 23, further comprising, prior to step (a), isolating the Tcell, the NK cell, or themacrophage, from an individual. 25. The method of clause 24, wherein the T cell, the NK cell, or the macrophage, is isolated from the patient. 26. Themethod of clause 23, wherein the Tcell is a tumor-infiltrating T lymphocyte, aCD4+Tcell, aCD8+Tcell, or the combination thereof. 27. The method of any one of clauses 22‑26, wherein the cancer is a hematological cancer. 28. The method of clause 27, wherein the hematological cancer is a GPRC5D-expressing B cell cancer. 29. The method of clause 28, wherein the GPRC5D-expressing B cell cancer is multiple myeloma.

[0157] Thepresent disclosure is not to be limited in scope by the specific embodiments describedwhich are intendedas 30 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 single illustrations of individual aspects of the disclosure, and any compositions or methods which are functionally equivalent arewithin the scopeof this disclosure. It will be apparent to those skilled in the art that variousmodifications and variations can be made in the methods and compositions of the present disclosure without departing from the spirit or scope of the disclosure. Thus, it is intended that the present disclosure cover the modifications and variations of this disclosure provided they come within the scope of the appended claims and their equivalents.

[0158] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. Claims 1. An antibody or antigen-binding fragment thereof having binding specificity to a human G-protein-coupled receptor family C group 5 member D (GPRC5D) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region (VH) comprising heavy chain complementarity determining regions CDRH1, CDRH2, and CDRH3 and a light chain variable region (VL) comprising complementarity determining regionsCDRL1,CDRL2, andCDRL3, and wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise: (a) the amino acid sequences of SEQ ID NO:42‑47; (b) the amino acid sequences of SEQ ID NO:29‑34; (c) the amino acid sequences of SEQ ID NO:54‑59; or (d) the amino acid sequences of SEQ ID NO:68‑73. 2. The antibody or antigen-binding fragment thereof of claim 1, wherein theCDRH1, CDRH2, CDRH3,CDRL1,CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:42‑47, optionally wherein theVHcomprisesanaminoacidsequenceselected fromthegroupconsistingofSEQIDNO:9and48‑50,and the VL comprises an amino acid sequence selected from the group consisting of SEQ IDNO:10 and 51‑53, or the VH comprises the amino acid sequence of SEQ ID NO:48, and the VL comprises the amino acid sequence of SEQ ID NO:51. 3. The antibody or antigen-binding fragment thereof of claim 1, wherein theCDRH1, CDRH2, CDRH3,CDRL1,CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:29‑34, optionally wherein theVHcomprisesanaminoacidsequenceselected fromthegroupconsistingofSEQIDNO:7and35‑37,and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:8 and 38‑41, or the VH comprises the amino acid sequence of SEQ ID NO:35, and the VL comprises the amino acid sequence of SEQ ID NO:38. 4. The antibody or antigen-binding fragment thereof of claim 1, wherein theCDRH1, CDRH2, CDRH3,CDRL1,CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:54‑59, optionally wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ IDNO:61‑64, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:16 and 65‑67, or the VH comprises the amino acid sequence of SEQ ID NO:61, and the VL comprises the amino acid sequence of SEQ ID NO:65, or the VH comprises the amino acid sequence of SEQ ID NO:63, and the VL comprises the amino acid sequence of SEQ ID NO:65. 5. The antibody or antigen-binding fragment thereof of claim 1, wherein theCDRH1, CDRH2, CDRH3,CDRL1,CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:68‑73, optionally wherein theVHcomprisesanaminoacidsequenceselected fromthegroupconsistingofSEQIDNO:1and74‑79,and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2 and 80‑86, or the VH comprises the amino acid sequence of SEQ ID NO:76, and the VL comprises the amino acid sequence of SEQ ID NO:82, or the VH comprises the amino acid sequence of SEQ ID NO:77, and the VL comprises the amino acid sequence of SEQ ID NO:82. 6. The antibody or antigen-binding fragment thereof of any one of claims 1‑5, wherein said antibody or antigen-binding fragment thereof is ADCC-competent. 7. The antibody or antigen-binding fragment thereof of any one of claims 1‑5, further comprising a cytotoxic drug conjugated to the antibody or fragment thereof. 31 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 8. The antibody or antigen-binding fragment thereof of claim 7, wherein the cytotoxin is a maytansinoid or an auristatin, optionallywherein the cytotoxin is selected from the group consisting ofmaytansine,N2’-deacetyl-N2’‑(3-mercapto-l- oxopropyl) or N2’-deacetyl-N2’‑(3-mercapto-l-oxopropyl)‑maytansine (DM1); monomethylauristatin-D (MMAD); monomethyl auristatin-E (MMAE); and monomethylauristatin-F (MMAF). 9. Abispecific antibody comprising the antigen-binding fragment of any one of claims 1‑5, and a second antigen-binding fragment having specificity toasecond target protein, optionallywherein the second target protein is selected from the group consisting of CD3, CD16, CD19, CD28, CD64 and 4‑1BB, such as CD3. 10. An antibody or antigen-binding fragment thereof, or a bispecific antibody for use in a method of treating cancer in a patient in need thereof,wherein theantibodyorantigen-binding fragment thereof is asdefined inanyoneof claims1‑8, and the bispecific antibody is as defined in claim 9. 11. The antibody or antigen-binding fragment thereof, or the bispecific antibody for use according to claims 10, wherein the cancer is a hematological cancer, optionally wherein the hematological cancer is a GPRC5D-expressing B cell cancer. 12. The antibody or antigen-binding fragment thereof, or the bispecific antibody for use according to claims 11, wherein the GPRC5D-expressing B cell cancer is multiple myeloma. 32 EP 4 670 724 A2 5 10 15 20 25 30 35 40 45 50 55 33 EP 4 670 724 A2 34 EP 4 670 724 A2 35 EP 4 670 724 A2 36 EP 4 670 724 A2 37 EP 4 670 724 A2 38 EP 4 670 724 A2 39 EP 4 670 724 A2 40 EP 4 670 724 A2 41 EP 4 670 724 A2 42 EP 4 670 724 A2 43 EP 4 670 724 A2 44 EP 4 670 724 A2 45 EP 4 670 724 A2 46 EP 4 670 724 A2 47 EP 4 670 724 A2 48 EP 4 670 724 A2 49 EP 4 670 724 A2 50 EP 4 670 724 A2 51 EP 4 670 724 A2 REFERENCES CITED IN THE DESCRIPTION This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard. Patent documents cited in the description • US 5892019 A

[0022] • US 6150584 A

[0093] • US 6458592 B

[0093] • US 6420140 B

[0093] Non-patent literature cited in the description • HAMERS-CASTERMAN et al. Nature, 1993, vol. 363, 446-448

[0027] • KABAT, E. et al. Sequences of Proteins of Immuno- logical Interest. U.S. Department of Health and Human Services, 1983

[0028] • CHOTHIA ; LESK. J. Mol. Biol., 1987, vol. 196, 901- 917

[0028] • KABAT et al. Sequences of Proteins of Immunolo- gical Interest. U.S. Dept. of Health and Human Services, 1983

[0029]

[0038] • CHOTHIA et al. J. Mol. Biol., 1987, vol. 196, 901-917

[0029] • KABATet al.Sequence of Proteins of Immunological Interest. U.S. Dept. of Health and Human Services, 1983

[0030] • ROUX et al. J. Immunol, 1998, vol. 161, 4083

[0039] • Monoclonal Antibodies For Immunotargeting Of Drugs InCancer Therapy. ARNONet al.Monoclonal Antibodies And Cancer Therapy. Alan R. Liss, Inc, 1985, 243-56

[0088] • Antibodies For Drug Delivery. HELLSTROM et al. Controlled Drug Delivery. Marcel Dekker, Inc., 1987, 623-53

[0088] • Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review. THORPE et al. Monoclonal Antibodies ’84: Biological And Clinical Applications. 1985, 475-506

[0088] • Analysis, Results, And Future Prospective Of The TherapeuticUseOfRadiolabeledAntibody InCancer Therapy. Monoclonal Antibodies For Cancer Detec- tion And Therapy. Academic Press, 1985, 303-16

[0088] • THORPE et al. The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates. Immunol. Rev., 1982, vol. 52, 119-58

[0088] (19) *EP004670724A3* (11) EP 4 670 724 A3 (12) EUROPEAN PATENT APPLICATION (88) Date of publication A3: 01.04.2026 Bulletin 2026 / 14 (43) Date of publication A2: 31.12.2025 Bulletin 2026 / 01 (21) Application number: 25209453.7 (22) Date of filing: 05.01.2022 (51) International Patent Classification (IPC): C07K 16 / 28 (2006.01) A61K 39 / 395 (2006.01) A61P 35 / 00 (2006.01) A61K 35 / 17 (2025.01) A61K 47 / 68 (2017.01) A61K 35 / 15 (2025.01) (52) Cooperative Patent Classification (CPC): A61P 35 / 00; A61K 35 / 15; A61K 35 / 17; A61K 47 / 68031; A61K 47 / 6849; C07K 16 / 28; A61K 2039 / 505; A61K 2039 / 54; A61K 2039 / 545; C07K 2317 / 24; C07K 2317 / 56; C07K 2317 / 565; C07K 2317 / 732; C07K 2317 / 76; C07K 2317 / 77; (Cont.) (84) Designated Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR Designated Extension States: BA ME Designated Validation States: MA TN (30) Priority: 05.01.2021 PCT / CN2021 / 070314 (62) Document number(s) of the earlier application(s) in accordance with Art. 76 EPC: 22736532.7 / 4 274 851 (71) Applicant: Lanova Medicines Development Co., Ltd. Shanghai 201315 (CN) (72) Inventors: • LI, Runsheng Shanghai 201315 (CN) • HUANG, Wentao Shanghai 201315 (CN) (74) Representative: AstraZeneca Intellectual Property Eastbrook House Shaftesbury Road Cambridge CB2 8BF (GB) (54) ANTI‑GPRC5D MONOCLONAL ANTIBODIES AND USES THEREOF (57) Provided are antibodies or fragment thereof havingbinding specificity to the humanGPRC5Dprotein. The antibodies or fragment thereof are capable of target- ing cancer cells expressing GPRC5D, and thus can be used to treat the cancer, in particular hematological can- cer. EP 4 67 0 72 4 A 3 Processed by Luminess, 75001 PARIS (FR) (Cont. next page) (52) Cooperative Patent Classification (CPC): (Cont.) C07K 2317 / 92 2 EP 4 670 724 A3 3 EP 4 670 724 A3 5 10 15 20 25 30 35 40 45 50 55 4 EP 4 670 724 A3 5 10 15 20 25 30 35 40 45 50 55 5 EP 4 670 724 A3 5 10 15 20 25 30 35 40 45 50 55 摘要 提供了对人GPRC5D蛋白具有结合特异性的抗体或其片段。该抗 体或其片段能够靶向表达GPRC5D的癌细胞，因此可用于治疗癌症， 特别是血液系统癌症。

Claims

1. An antibody or antigen-binding fragment thereof having binding specificity to a human G-protein-coupled receptor family C group 5 member D (GPRC5D) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region (VH) comprising heavy chain complementarity determining regions CDRH1, CDRH2, and CDRH3 and a light chain variable region (VL) comprising complementarity determining regions CDRL1, CDRL2, and CDRL3, and wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise: (a) the amino acid sequences of SEQ ID NO:42-47; (b) the amino acid sequences of SEQ ID NO:29-34; (c) the amino acid sequences of SEQ ID NO:54-59; or (d) the amino acid sequences of SEQ ID NO:68-73.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:42-47, optionally wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:9 and 48-50, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:10 and 51-53, or the VH comprises the amino acid sequence of SEQ ID NO:48, and the VL comprises the amino acid sequence of SEQ ID NO:51.

3. The antibody or antigen-binding fragment thereof of claim 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:29-34, optionally wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:7 and 35-37, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:8 and 38-41, or the VH comprises the amino acid sequence of SEQ ID NO:35, and the VL comprises the amino acid sequence of SEQ ID NO:38.

4. The antibody or antigen-binding fragment thereof of claim 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:54-59, optionally wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:61-64, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:16 and 65-67, or the VH comprises the amino acid sequence of SEQ ID NO:61, and the VL comprises the amino acid sequence of SEQ ID NO:65, or the VH comprises the amino acid sequence of SEQ ID NO:63, and the VL comprises the amino acid sequence of SEQ ID NO:65.

5. The antibody or antigen-binding fragment thereof of claim 1, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, respectively, comprise the amino acid sequences of SEQ ID NO:68-73, optionally wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1 and 74-79, and the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2 and 80-86, or the VH comprises the amino acid sequence of SEQ ID NO:76, and the VL comprises the amino acid sequence of SEQ ID NO:82, or the VH comprises the amino acid sequence of SEQ ID NO:77, and the VL comprises the amino acid sequence of SEQ ID NO:82.

6. The antibody or antigen-binding fragment thereof of any one of claims 1-5, wherein said antibody or antigen-binding fragment thereof is ADCC-competent.

7. The antibody or antigen-binding fragment thereof of any one of claims 1-5, further comprising a cytotoxic drug conjugated to the antibody or fragment thereof.

8. The antibody or antigen-binding fragment thereof of claim 7, wherein the cytotoxin is a maytansinoid or an auristatin, optionally wherein the cytotoxin is selected from the group consisting of maytansine, N2'-deacetyl-N2'-(3-mercapto-l-oxopropyl) or N2'-deacetyl-N2'-(3-mercapto-l-oxopropyl)-maytansine (DM1); monomethylauristatin-D (MMAD); monomethyl auristatin-E (MMAE); and monomethylauristatin-F (MMAF).

9. A bispecific antibody comprising the antigen-binding fragment of any one of claims 1-5, and a second antigen-binding fragment having specificity to a second target protein, optionally wherein the second target protein is selected from the group consisting of CD3, CD16, CD19, CD28, CD64 and 4-1BB, such as CD3.

10. An antibody or antigen-binding fragment thereof, or a bispecific antibody for use in a method of treating cancer in a patient in need thereof, wherein the antibody or antigen-binding fragment thereof is as defined in any one of claims 1-8, and the bispecific antibody is as defined in claim 9.

11. The antibody or antigen-binding fragment thereof, or the bispecific antibody for use according to claims 10, wherein the cancer is a hematological cancer, optionally wherein the hematological cancer is a GPRC5D-expressing B cell cancer.

12. The antibody or antigen-binding fragment thereof, or the bispecific antibody for use according to claims 11, wherein the GPRC5D-expressing B cell cancer is multiple myeloma.

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