Anti-CCR8 antibodies and uses thereof

Abstract

The present invention discloses an anti-CCR8 antibody or an antigen binding portion thereof; a pharmaceutical composition comprising the anti-CCR8 antibody or an antigen binding portion thereof and a pharmaceutically acceptable carrier; a method of treating a disease or condition in a subject in need thereof using the anti-CCR8 antibody or antigen-binding portion or the pharmaceutical composition; and the use of the anti-CCR8 antibody or the antigen-binding portion thereof in the manufacture of a medicament for the treatment of a disease or disorder.

Description

Background Art

[0001] Immunotherapy is a rapidly developing and very promising treatment for many forms of cancer, with many recent successes. However, some patients have limited or no response to current immunotherapies, and others experience relapse after an initial response.

[0002] The human immune system includes checks and balances to prevent an overactive immune system from harming the body. Regulatory T cells (Tregs) play an important role in maintaining immune system function by suppressing immune responses. However, when Tregs infiltrate tumors, they can block immune responses to tumors. Therefore, there is a need to develop a more effective method to selectively target tumor-infiltrating Tregs. Since CCR8 prefers to be expressed on tumor-infiltrating Tregs, targeting CCR8 may be an effective therapeutic strategy to specifically regulate tumor-infiltrating Tregs in the tumor microenvironment (TME) and enhance anti-tumor immunity. Summary of the invention

[0003] The present disclosure provides anti-CCR8 antibodies that specifically bind to CCR8 and effectively inhibit tumor growth, and thus can be used to prevent or treat various cancers.

[0004] In a first aspect, the present disclosure provides an anti-CCR8 antibody or an antigen-binding portion thereof, which comprises a heavy chain CDR1 having an amino acid sequence shown in any one of SEQ ID NOs: 1-2, 56-60, 62, a heavy chain CDR2 having an amino acid sequence shown in any one of SEQ ID NOs: 3, 63-67, 69-73, a heavy chain CDR3 having an amino acid sequence shown in any one of SEQ ID NOs: 4-5, 74-78, 80-85, a light chain CDR1 having an amino acid sequence shown in any one of SEQ ID NOs: 6-11, 86-90, 92-93, a light chain CDR2 having an amino acid sequence shown in any one of SEQ ID NOs: 12, 94-98, 100-102, and a light chain CDR3 having an amino acid sequence shown in any one of SEQ ID NOs: 13-14, 103-107, 109-113.

[0005] In a second aspect, the present disclosure provides a pharmaceutical composition comprising an anti-CCR8 antibody or antigen-binding portion thereof according to the present disclosure and a pharmaceutically acceptable carrier.

[0006] In a third aspect, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject an anti-CCR8 antibody or antigen-binding portion according to the present disclosure or a pharmaceutical composition according to the present disclosure.

[0007] In a fourth aspect, the present disclosure provides use of an anti-CCR8 antibody or antigen-binding portion thereof according to the present disclosure in the manufacture of a medicament for treating a disease or disorder. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 Binding titration of anti-CCR8 mAbs ChH2, H2A, H2B, H2C, H2G and H2I to TALL-1 cells with endogenous human CCR8 on the cell surface is shown, where the X-axis represents the concentration of anti-CCR8 mAb and the Y-axis represents the mean fluorescence intensity (MFI).

[0009] Figure 2 Binding titration of anti-CCR8 mAbs ABC047-B1.IgG1.SDIE, ABC048-B1.hu11.IgG1.SDIE, ABC049-B1.hu12.IgG1.SDIE, ABC050-B1.hu13.IgG1.SDIE, ABC051-B1.hu14.IgG1.SDIE to 293T cells overexpressing human CCR8 is shown.

[0010] Figure 3 Binding titration of anti-CCR8 mAbs ABC025-H2G.SDIE, ABC047-B1.IgG1.SDIE, ABC137-H2.hu12.IgG1.SDIE and ABC138-H2.hu13.IgG1.SDIE to 293T cells overexpressing human CCR8 is shown, where the X-axis represents the concentration of anti-CCR8 mAb and the Y-axis represents the MFI.

[0011] Figure 4 Binding titration of anti-CCR8 mAbs ABC025-H2G.SDIE, ABC048-B1.hu11.IgG1.SDIE, ABC138-H2.hu13.IgG1.SDIE, ABC078.AF and ABC084.AF to activated Treg cells derived from human PBMCs is shown.

[0012] Figure 5Binding titration of anti-CCR8 mAbs ABC814, ABC815, ABC816, ABC817, ABC818, ABC819, ABC820, ABC821, ABC822, ABC823, ABC531, ABC138-H2.hu13.IgG1.SDIE, ABC080.chH2.SDIE, ABC026, and ABC078 to 293T cells overexpressing human or cynomolgus monkey CCR8 and Hut78 cell line with endogenous CCR8 are shown, where the X-axis represents the logarithm of the anti-CCR8 mAb concentration and the Y-axis represents the GeoMean.

[0013] Figure 6 Titration of anti-CCR8 mAbs ABC138-H2.hu13.IgG1.SDIE, ABC025-H2G.SDIE and ABC080.chH2.SDIE on human PBMC-derived activated Treg cells is shown, with the X-axis representing antibody concentration and the Y-axis representing MFI.

[0014] Figure 7 The calcium flux measurement of CHEM1-CCR8 cells with high expression of human CCR8 on the cell surface is shown, wherein the X-axis represents time (s) and the Y-axis represents the normalized chemotaxis ratio calculated by the following formula: [F (fluorescence signal) - Fmin (minimum fluorescence signal representing the baseline signal)] / Fmin. Different antibodies were used at a concentration of 66.7 nM.

[0015] Figure 8 Shown is a calcium flux dose titration assay of CHEM1-CCR8 cells with high human CCR8 expression on the cell surface, where the X-axis represents the concentration of anti-CCR8 mAb and the Y-axis represents the normalized chemotactic signal calculated by the following formula: Maximum (maximum fluorescence signal achieved within 90 min after CCL1 injection) - Minimum (minimum fluorescence signal representing the baseline signal).

[0016] Fig. 9 ChH2-AF or ChB1-AF mediated ADCC killing of TALL-1 cells is shown.

[0017] Fig.10 Shown is ADCC killing of Hut78 cells mediated by ABC024-H2G, ABC024-H2G.AF, and ABC025-H2G.SDIE at fixed concentrations.

[0018] Fig.11 Shown is ADCC killing of Hut78 cells mediated by ABC024-H2G.AF and ABC025-H2G.SDIE.

[0019] Fig.12 ADCC killing of Jurkat-cynoCCR8 cells mediated by ABC089, ABC078, ABC138-H2.hu13.IgG1.SDIE, ABC822, and ABC1183 are shown.

[0020] Fig.13 ADCC killing of Jurkat-hCCR8 mediated by ABC089, ABC078, ABC138-H2.hu13.IgG1.SDIE, ABC822, and ABC1183 is shown.

[0021] Fig.14 ADCC killing of Hut78 cells mediated by ABC089, ABC078.AF, ABC138-H2.hu13.IgG1.SDIE, ABC822, and ABC1183 are shown. DETAILED DESCRIPTION

[0022] As used in this application, unless otherwise expressly provided herein, each of the following terms shall have the meaning set forth below. Additional definitions are set forth throughout the application.

[0023] It should be noted that "a" or "an" entity refers to one or more of the entity; for example, "an antibody" is understood to represent one or more antibodies. Similarly, the terms "a", "one or more", and "at least one" are used interchangeably herein.

[0024] Furthermore, "and / or" when used herein should be understood as a specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and / or" as used in phrases such as "A and / or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0025] The term "about" is used herein to mean approximately, roughly, approximately, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values ​​set forth above.

[0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure relates.

[0027] Units, prefixes and symbols are all expressed in their SI accepted form. Numerical ranges include numbers that define the range. Unless otherwise indicated, nucleotide sequences are written from left to right in a 5' to 3' direction. Amino acid sequences are written from left to right in an amino to carboxyl direction. The headings provided herein are not limitations of the various aspects of the disclosure, which can be obtained by reference to the specification as a whole. Therefore, the terms defined immediately below are more fully defined by reference to the specification in its entirety.

[0028] As used herein, the term "amount" or "level" is used in the broadest sense and refers to the amount, concentration or abundance of a substance (e.g., a metabolite, a small molecule, a protein, an mRNA, a marker). When referring to a metabolite or a small molecule (e.g., a drug), the terms "amount", "level" and "concentration" are often used interchangeably and generally refer to the detectable amount in a biological sample. "Elevated levels" or "increased levels" refer to an increase in the amount, concentration or abundance of a substance in a sample relative to, for example, a control sample or internal control from one or more individuals who do not suffer from the disease or disorder (e.g., cancer). In some respects, the level of material (for example, drug) in sample increases, refers to as determined by technology known in the art (for example, HPLC), relative to the amount of material in control sample, the amount of this material increases by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%. " reduced level " refers to the quantity, concentration or abundance of material (for example, drug) in individual relative to the reduction of for example from one or more individual controls of not suffering from this disease or obstacle (for example, cancer) or internal controls. In some respects, the level of reduction is seldom or does not have detectable quantity, concentration or abundance. In some aspects, a decrease in the level of a substance (e.g., a drug) in a sample refers to a decrease in the amount of the substance by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% relative to the amount of the substance in a control sample as determined by techniques known in the art (e.g., HPLC).

[0029] When referring to a protein, mRNA or marker (such as those described herein), the term "level of expression" or "expression level" is generally used interchangeably and generally refers to the detectable amount of a protein, mRNA or marker in a biological sample. In some aspects, the detectable amount or detectable level of a protein, mRNA or marker is associated with the likelihood of responding to an agent (such as those described herein). "Expression" generally refers to the process of converting the information contained in a gene into a structure (e.g., a protein marker, e.g., PD-L1) present and functioning in a cell. Therefore, as used herein, "expression" may refer to transcription into a polynucleotide, translation into a polypeptide, or even polynucleotide and / or polypeptide modification (e.g., post-translational modification of a polypeptide). Fragments of transcribed polynucleotides, translated polypeptides or polynucleotide and / or polypeptide modifications (e.g., post-translational modification of a polypeptide) should also be considered to be expressed, whether they are derived from transcripts generated by alternative splicing or degraded transcripts, or from post-translational processing of a polypeptide, e.g., by proteolysis. "Expressed genes" include genes that are transcribed into polynucleotides as mRNA and then translated into polypeptides, and also include genes that are transcribed into RNA but not translated into polypeptides (e.g., transport and ribosomal RNA). "Elevated expression", "elevated expression level" or "elevated level" refers to the expression of a substance in a sample relative to, for example, a control sample or internal control of one or more individuals who do not suffer from the disease or disorder (e.g., cancer) or an increased level. In some aspects, the expression of a substance (e.g., a protein marker, such as PD-L1) in a sample is increased, and refers to, as determined by techniques known in the art (e.g., FACS), relative to the amount of the substance in the control sample, the amount of the substance increases by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. "Reduced expression", "reduced expression level" or "reduced level" refers to the expression reduction or level reduction of a substance (e.g., protein marker) in an individual relative to a control or internal control of one or more individuals who do not suffer from the disease or disorder (e.g., cancer). In some aspects, reduced expression is very little or no expression. In some aspects, the expression reduction of a substance (e.g., protein marker) in a sample refers to, as determined by techniques known in the art (e.g., FACS), relative to the amount of the substance in a control sample, the amount of the substance is reduced by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0030] As used herein, the term "antibody" refers to a complete antibody comprising two light chain polypeptides and two heavy chain polypeptides. Complete antibodies include different antibody isotypes, including IgM, IgG, IgA, IgD and IgE antibodies. The term "antibody" includes polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, primatized antibodies, deimmunized antibodies and fully human antibodies. Antibodies can be made in any of a variety of species or derived therefrom, for example, mammals such as humans, non-human primates (e.g., orangutans, baboons or chimpanzees), horses, cattle, pigs, sheep, goats, dogs, cats, rabbits, guinea pigs, gerbils, hamsters, rats and mice. Antibodies can be purified or recombinant antibodies.

[0031] As used herein, the terms "antibody fragment", "antigen binding fragment", "antigen binding portion", "antigen binding protein" or similar terms refer to a fragment / portion of an antibody that retains the ability to bind to a target antigen (e.g., CCR8) and inhibit the activity of the target antigen. Such fragments include, for example, single-chain antibodies, single-chain Fv fragments (scFv), Fd fragments, Fab fragments, Fab' fragments, or F(ab')2 fragments. A scFv fragment is a single polypeptide chain that includes the heavy chain and light chain variable regions of the antibody from which the scFv is derived. In addition, intrabodies, minibodies, triabodies, and diabodies are also included in the definition of antibodies, and they are suitable for use in the methods described herein.

[0032] As known in the art, "affinity" is a measure of the tightness with which a particular ligand binds to its partner. Affinity can be measured in different ways. In some embodiments, affinity is measured by quantitative determination. In some such embodiments, the binding partner concentration can be fixed to exceed the ligand concentration to simulate physiological conditions. Alternatively or additionally, in some embodiments, the binding partner concentration and / or ligand concentration can be changed. In some such embodiments, affinity can be compared with a reference under comparable conditions (e.g., concentration). "Affinity maturation" (or "affinity matured antibody"): as used herein, refers to an antibody with one or more changes in one or more of its CDRs, which increases the affinity of the antibody to the antigen compared to the parent antibody without those changes. In some embodiments, affinity matured antibodies will have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies can be produced by any of a variety of procedures known in the art.

[0033] As used herein, the term "administering" typically refers to administering a composition to a subject or system to achieve delivery of the composition agent or the agent contained in the composition. Those of ordinary skill in the art will understand that a variety of routes can be used for administering to a subject (e.g., human) where appropriate. For example, in some embodiments, administration may be by ocular, oral, parenteral, topical, and other routes. In some specific embodiments, administration may be by bronchial (e.g., by bronchial instillation), oral, skin (which may be or include, for example, one or more of the routes of surface to dermis, intradermal, interdermal, transdermal, etc.), enteral, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, in a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, and other routes. In some embodiments, administration may involve only a single administration. In some embodiments, administration may involve the use of a fixed number of doses. In some embodiments, administration may involve intermittent (e.g., multiple doses separated in time) and / or regular (e.g., individual doses separated by a common time period) administration. In some embodiments, administration may involve continuous administration (e.g., perfusion) for at least one selected time period.

[0034] Amino Acid: As used herein, in its broadest sense refers to any compound and / or substance that can be incorporated into a polypeptide chain, for example, by forming one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N–C(H)(R)–COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a non-natural amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. "Non-standard amino acid" refers to any amino acid other than a standard amino acid, whether it is prepared synthetically or obtained from a natural source. In some embodiments, amino acids (including carboxyl-terminal and / or amino-terminal amino acids in polypeptides) may contain structural modifications compared to the general structure described above. For example, in some embodiments, amino acids may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., substitution of an amino group, a carboxylic acid group, one or more protons, and / or a hydroxyl group) compared to the general structure. In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide comprising a modified amino acid, compared to a polypeptide comprising an otherwise identical unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide comprising a modified amino acid, compared to a polypeptide comprising an otherwise identical unmodified amino acid. As will be apparent from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments, an amino acid may be used to refer to an amino acid residue of a polypeptide.

[0035] As used herein, "cancer antigen" or "tumor antigen" refers to (i) tumor-specific antigens, (ii) tumor-associated antigens, (iii) cells expressing tumor-specific antigens, (iv) cells expressing tumor-associated antigens, (v) embryonic antigens on tumors, (vi) autologous tumor cells, (vii) tumor-specific membrane antigens, (viii) tumor-associated membrane antigens, (ix) growth factor receptors, (x) growth factor ligands, and (xi) any other type of antigen or antigen-presenting cell or material associated with cancer.

[0036] As used herein, the term "cancer-specific immune response" refers to an immune response induced by the presence of a tumor, cancer cell, or cancer antigen. In some aspects, the response includes the proliferation of cancer antigen-specific lymphocytes. In some aspects, the response includes the expression and upregulation of antibodies and T cell receptors, and the formation and release of lymphokines, chemokines, and cytokines. Both the innate and acquired immune systems interact to initiate an antigenic response to a tumor, cancer cell, or cancer antigen. In some aspects, the cancer-specific immune response is a T cell response.

[0037] As used herein, the term "CCR8" or "CC chemokine receptor type 8" refers to a G protein-coupled receptor.

[0038] As used herein, the term "combination therapy" refers to situations in which a subject is exposed to two or more treatment regimens (eg, two or more therapeutic agents) simultaneously.

[0039] As used herein, "CDR" refers to the complementarity determining region within the variable region of an antibody. Each variable region of the heavy chain and light chain has three CDRs, which are named CDR1, CDR2, and CDR3 for each variable region. A "group of CDRs" or "CDR group" refers to a group of three or six CDRs that appear in a single variable region that is capable of binding an antigen or a plurality of CDRs of homologous heavy and light chain variable regions that are capable of binding an antigen. Certain systems for defining CDR boundaries have been established in the art (e.g., Kabat, Chothia, etc.); those skilled in the art understand the differences between these systems and are able to understand CDR boundaries to the extent necessary to understand and practice the claimed invention.

[0040] As used herein, the term "competition" when used in the context of antigen binding proteins (e.g., immunoglobulins, antibodies, or antigen binding fragments thereof) that compete for binding to the same epitope refers to an interaction between antigen binding proteins as determined by an assay (e.g., a competitive binding assay; a cross-blocking assay) in which a test antigen binding protein (e.g., a test antibody) inhibits (e.g., reduces or blocks) specific binding of a reference antigen binding protein (e.g., a reference antibody) to a common antigen (e.g., CCR8 or a fragment thereof).

[0041] As used herein, the term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The term "therapeutically effective dose" is defined as an amount sufficient to cure or at least partially arrest the disease and its complications in the respective patient. The effective amount for this purpose depends on the severity of the disorder being treated and the overall state of the patient's own immune system.

[0042] As used herein, "bispecific antibody" refers to a bispecific adhesive in which at least one, typically two, binding moieties are or comprise antibody components. A variety of different bispecific antibody structures are known in the art. In some embodiments, each binding moiety in a bispecific antibody as or comprising an antibody component includes a VH and / or VL region; in some such embodiments, the VH and / or VL region is those found in a specific monoclonal antibody. In some embodiments, wherein the bispecific antibody comprises two antibody component binding moieties, each comprising a VH and / or VL region from a different monoclonal antibody. In some embodiments, in the case where the bispecific antibody comprises two antibody component binding moieties, wherein one of the two antibody component binding moieties comprises an immunoglobulin molecule having a VH and / or VL region comprising a CDR from a first monoclonal antibody, and one of the two antibody component binding moieties comprises an antibody fragment (e.g., Fab, F(ab'), F(ab')2, Fd, Fv, dAB, scFv, etc.) having a VH and / or VL region comprising a CDR from a second monoclonal antibody.

[0043] The term "human antibody", as used herein, includes antibodies having variable and constant regions (if present) of human germline immunoglobulin sequences.

[0044] As used herein, the term "humanization" refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody are replaced by corresponding amino acids derived from human immunoglobulins. In some embodiments of humanized forms of antibodies, some, most or all of the amino acids outside the CDR domains have been replaced by amino acids from human immunoglobulins, while some, most or all of the amino acids within one or more CDR regions are unchanged. A small amount of addition, deletion, insertion, substitution or modification of amino acids is allowed as long as they do not eliminate the ability of the antibody to bind to a specific antigen. "Humanized" antibodies retain antigenic specificity similar to the original antibody.

[0045] "Chimeric antibody" refers to an antibody whose variable region is derived from one species and whose constant region is derived from another species, such as an antibody whose variable region is derived from a mouse antibody and whose constant region is derived from a human antibody.

[0046] As used herein, the term "heterologous antibody" is defined in relation to a transgenic non-human organism that produces such an antibody. The term refers to an antibody having an amino acid sequence or encoding nucleic acid sequence that corresponds to an amino acid sequence or encoding nucleic acid sequence found in an organism that is not composed of the transgenic non-human animal, typically from a species other than the transgenic non-human animal.

[0047] As used herein, "antigen" refers to an agent that elicits an immune response; and / or (ii) an agent that binds to a T cell receptor (eg, when presented by an MHC molecule) or an antibody.

[0048] The terms "induce an immune response" and "enhance an immune response" are used interchangeably and refer to stimulating an immune response (ie, passive or adaptive) against a specific antigen. When used relative to inducing CDC or ADCC, the term "induce" refers to stimulating a specific direct cell killing mechanism.

[0049] As used herein, the term "immunogenic cell death" (also referred to as "immunogenic apoptosis") refers to a cell death pattern associated with the activation of one or more signal transduction pathways, which induces the release of pro-death expression and damage-associated molecular pattern (DAMP) molecules (e.g., adenosine triphosphate, ATP) of tumor cells, resulting in increased immunogenicity of tumor cells and death in an immunogenic manner (e.g., by phagocytosis). As used herein, the term "immunogenic cell death inducer" refers to a chemical, biological or pharmacological agent that induces an immunogenic cell death process, pathway or pattern.

[0050] The term "in vivo" refers to processes that occur within a living organism.

[0051] As used herein, the term "Kd" refers to the equilibrium dissociation constant of the binding reaction between an antibody and an antigen. The Kd value is a numerical representation of the ratio of the antibody dissociation rate (off-rate) constant (kd) to the antibody association rate (on-rate) constant (ka). The Kd value is inversely proportional to the binding affinity of the antibody to the antigen. The smaller the Kd value, the greater the affinity of the antibody to its antigen. Affinity is the strength of the binding of a single molecule to its ligand, typically measured and reported using the equilibrium dissociation constant (Kd), which is used to evaluate the strength of bimolecular interactions and rank them.

[0052] As used herein, the term "kd" (or "koff") refers to the dissociation rate constant for the dissociation of an antibody from an antibody / antigen complex. The kd value is a numerical representation of the fraction of the complex that decays or dissociates per second and is expressed in units of S -1 express.

[0053] As used herein, the term "ka" (or "kon") refers to the association rate constant for the association of an antibody with an antigen. The ka value is a numerical representation of the number of antibody / antigen complexes formed per second by an antibody and an antigen in a 1 molar (1 M) solution, expressed in units of M. -1 ·S -1 express.

[0054] As used herein, the term "monoclonal antibody" refers to an antibody that exhibits a single binding specificity and affinity for a particular epitope. Thus, the term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity and has a variable region and optionally a constant region derived from a human germline immunoglobulin sequence. In some aspects, human monoclonal antibodies are produced by a hybridoma comprising a B cell obtained from a transgenic non-human animal, e.g., a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell.

[0055] As used herein, the term "natural killer (NK) cell" refers to a type of cytotoxic lymphocyte. These cells are large, usually granular, non-T, non-B lymphocytes that can kill certain tumor cells and play an important role in natural immunity against viruses and other intracellular pathogens, as well as antibody-dependent cell-mediated cytotoxicity (ADCC).

[0056] As used herein, the term "antibody-dependent cellular toxicity" or "ADCC" refers to the phenomenon that target cells bound to an antibody are killed by immune effector cells. Without wishing to be bound by any particular theory, we observe that ADCC is typically understood to involve effector cells carrying Fc receptors (FcRs) that are able to recognize and subsequently kill target cells coated with antibodies (e.g., cells expressing specific antigens bound to the antibody on their surfaces). Effector cells that mediate ADCC may include immune cells, including but not limited to one or more of natural killer (NK) cells, macrophages, neutrophils, and eosinophils.

[0057] As used herein, the term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single or double-stranded form. Unless otherwise limited, the term encompasses nucleic acids containing known natural nucleotide analogs, which have binding properties similar to reference nucleic acids and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a specific nucleic acid sequence also implicitly encompasses variants and complementary sequences thereof that are conservatively modified and sequences that are clearly indicated. In particular, degenerate codon substitutions can be achieved by generating the following sequences, in which the third position of one or more selected (or all) codons is replaced by mixed bases and / or deoxyinosine residues. For arginine and leucine, the modification of the second base can also be conservative. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene.

[0058] The polynucleotide used herein can be composed of any polyribonucleotide or polydeoxynucleotide, which can be unmodified RNA or DNA or modified RNA or DNA. For example, the polynucleotide can be composed of single-stranded and double-stranded DNA, DNA mixed with single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA mixed with single-stranded and double-stranded regions, and a hybrid molecule of DNA and RNA containing single-stranded or more typically double-stranded or single-stranded and double-stranded regions. In addition, the polynucleotide can be composed of a triple-stranded region containing RNA or DNA or both RNA and DNA. The polynucleotide can also include one or more modified bases or a DNA or RNA backbone modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and rare bases such as inosine. DNA and RNA can be subjected to a variety of modifications; therefore, "polynucleotides" include chemical, enzymatic or metabolic modified forms.

[0059] As used herein, the term "patient" includes human and other mammalian subjects receiving prophylactic or therapeutic treatment.

[0060] As used generally herein, "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs and / or body fluids of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0061] As used herein, "pharmaceutically acceptable carrier" means, and includes, any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. that are physiologically compatible. These compositions may include pharmaceutically acceptable salts, for example, acid addition salts or base addition salts.

[0062] As used herein, the terms "polypeptide", "peptide" and "protein" are used interchangeably to refer to polymers of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding amino acid occurring in nature, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0063] As used herein, the term "preventing" when used in relation to a condition refers to reducing the frequency or delaying the onset of symptoms of a medical condition by administering the composition to a subject relative to a subject not receiving the composition.

[0064] As used herein, the terms "specific binding," "selective binding," "selectively binds," and "specifically binds" refer to the binding of an antibody to an epitope on a predetermined antigen.

[0065] As used herein, "epitope" includes any portion that is specifically recognized by an immunoglobulin (e.g., an antibody or a receptor) binding component. In some embodiments, an epitope is composed of a plurality of chemical atoms or groups on an antigen. In some embodiments, when an antigen adopts a related three-dimensional conformation, such chemical atoms or groups are exposed on the surface. In some embodiments, when an antigen adopts such a conformation, such chemical atoms or groups are physically close to each other in space. In some embodiments, when an antigen adopts an alternative conformation (e.g., linearized), at least some of such chemical atoms or groups are physically separated from each other.

[0066] As used herein, the term "T cell-mediated response" refers to any response mediated by T cells, including, but not limited to, effector T cells (e.g., CD8+ cells) and helper T cells (e.g., CD4+ cells). T cell-mediated responses include, for example, T cell cytotoxicity and proliferation.

[0067] As used herein, the terms "regulatory T cells," "T regulatory cells," "Tregs," or "Tregs" are used interchangeably herein and refer to T cell subsets that regulate the immune system, maintain tolerance to self-antigens, and prevent autoimmune diseases. Tregs have immunosuppressive effects, typically inhibiting or downregulating the induction and proliferation of effector T cells. Tregs are known to direct effector T cell lysis, support the formation of tolerogenic dendritic cells, support the formation of M2 macrophages, produce immunosuppressive metabolites and cytokines, serve as an IL-2 reservoir, and promote the formation of new vasculature. Although there are many types of Tregs, many Tregs express CD4 and FOXP3, and FOXP3 is used as a marker for Tregs in many cases.

[0068] As used herein, the term "therapeutically effective amount" or "therapeutically effective dose" or similar terms used herein means that amount of an agent that will elicit a desired biological or medical response (eg, improvement of one or more symptoms of cancer).

[0069] As used herein, the terms "treat", "treating" and "treatment" refer to therapeutic or preventive measures as described herein. These "treatment" methods are to administer human antibodies of the present disclosure to subjects in need of such treatment, such as subjects who need an enhanced immune response to a specific antigen or subjects who may eventually suffer from such a disorder, in order to prevent, cure, delay, improve one or more symptoms of the disorder or recurrent disorder, reduce its severity, or to prolong the survival of the subject beyond the expectation when not receiving such treatment.

[0070] As used herein, the term "tumor microenvironment" (or "cancer microenvironment", abbreviated TME) refers to the cellular environment or surrounding background in which a tumor or tumor neoplasm is located, including surrounding blood vessels and non-cancerous cells, including but not limited to immune cells, fibroblasts, bone marrow-derived inflammatory cells and lymphocytes. Signaling molecules and extracellular matrix also constitute TME. Tumors are closely related to the surrounding microenvironment and constantly interact with each other. Tumors can affect the microenvironment by releasing extracellular signals, promoting tumor angiogenesis and inducing peripheral immune tolerance, and immune cells in the microenvironment can affect the growth and evolution of tumor cells.

[0071] As used herein, the term "tumor" refers to an abnormal growth of cells or tissues. In some embodiments, a tumor may comprise precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and / or non-metastatic cells. In some embodiments, a tumor is associated with or is a manifestation of cancer. In some embodiments, a tumor may be a dispersed tumor or a liquid tumor. In some embodiments, a tumor may be a solid tumor.

[0072] As used herein, the term "solid tumor" refers to an abnormal mass of tissue that does not typically contain cysts or fluid areas. In some embodiments, a solid tumor may be benign; in some embodiments, a solid tumor may be malignant. One skilled in the art will appreciate that different types of solid tumors are typically named after the type of cells that form the solid tumor. Examples of solid tumors are carcinomas, lymphomas, and sarcomas.

[0073] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or an antigen-binding portion thereof, comprising a heavy chain CDR1 having an amino acid sequence shown in any one of SEQ ID NOs: 1-2, 56-60, 62, a heavy chain CDR2 having an amino acid sequence shown in any one of SEQ ID NOs: 3, 63-67, 69-73, a heavy chain CDR3 having an amino acid sequence shown in any one of SEQ ID NOs: 4-5, 74-78, 80-85, a light chain CDR1 having an amino acid sequence shown in any one of SEQ ID NOs: 6-11, 86-90, 92-93, a light chain CDR2 having an amino acid sequence shown in any one of SEQ ID NOs: 12, 94-98, 100-102, and a light chain CDR3 having an amino acid sequence shown in any one of SEQ ID NOs: 13-14, 103-107, 109-113.

[0074] In some aspects, in the anti-CCR8 antibody or antigen binding portion thereof:

[0075] 1) HCDR3 has the amino acid sequence shown in SEQ ID NO: 4, 74-78, 80-85; and

[0076] 2) LCDR1 has any one of the amino acid sequences shown in SEQ ID NOs: 7, 10-11, 86-90, 92-93.

[0077] In some aspects, in the anti-CCR8 antibody or antigen binding portion thereof:

[0078] 1) LCDR1 has any one of the amino acid sequences shown in SEQ ID NOs: 10-11, 86-90, 92-93; and

[0079] 2) LCDR3 has any one of the amino acid sequences shown in SEQ ID NOs: 14, 103-107, 109-113.

[0080] In some aspects, in the anti-CCR8 antibody or antigen binding portion thereof, LCDR1 has the amino acid sequence shown in SEQ ID NO: 10, 86-90, 92-93.

[0081] In some aspects, in the anti-CCR8 antibody or antigen binding portion thereof:

[0082] 1) HCDR1 has the amino acid sequence shown in SEQ ID NO: 1, HCDR2 has the amino acid sequence shown in SEQ ID NO: 3, HCDR3 has the amino acid sequence shown in SEQ ID NO: 4, LCDR1 has the amino acid sequence shown in SEQ ID NO: 6, LCDR2 has the amino acid sequence shown in SEQ ID NO: 12, and LCDR3 has the amino acid sequence shown in SEQ ID NO: 13;

[0083] 2) HCDR1 has the amino acid sequence shown in SEQ ID NO:2, HCDR2 has the amino acid sequence shown in SEQ ID NO:3, HCDR3 has the amino acid sequence shown in SEQ ID NO:5, LCDR1 has the amino acid sequence shown in SEQ ID NO:11, LCDR2 has the amino acid sequence shown in SEQ ID NO:12, and LCDR3 has the amino acid sequence shown in SEQ ID NO:14;

[0084] 3) HCDR1 has the amino acid sequence shown in SEQ ID NO: 1, HCDR2 has the amino acid sequence shown in SEQ ID NO: 3, HCDR3 has the amino acid sequence shown in SEQ ID NO: 4, LCDR1 has the amino acid sequence shown in SEQ ID NO: 7, LCDR2 has the amino acid sequence shown in SEQ ID NO: 12, and LCDR3 has the amino acid sequence shown in SEQ ID NO: 13;

[0085] 4) HCDR1 has the amino acid sequence shown in SEQ ID NO:2, HCDR2 has the amino acid sequence shown in SEQ ID NO:3, HCDR3 has the amino acid sequence shown in SEQ ID NO:5, LCDR1 has the amino acid sequence shown in SEQ ID NO:8, LCDR2 has the amino acid sequence shown in SEQ ID NO:12, and LCDR3 has the amino acid sequence shown in SEQ ID NO:14;

[0086] 5) HCDR1 has the amino acid sequence shown in SEQ ID NO:2, HCDR2 has the amino acid sequence shown in SEQ ID NO:3, HCDR3 has the amino acid sequence shown in SEQ ID NO:5, LCDR1 has the amino acid sequence shown in SEQ ID NO:9, LCDR2 has the amino acid sequence shown in SEQ ID NO:12, and LCDR3 has the amino acid sequence shown in SEQ ID NO:14;

[0087] 6) HCDR1 has the amino acid sequence shown in SEQ ID NO:2, HCDR2 has the amino acid sequence shown in SEQ ID NO:3, HCDR3 has the amino acid sequence shown in SEQ ID NO:5, LCDR1 has the amino acid sequence shown in SEQ ID NO:10, LCDR2 has the amino acid sequence shown in SEQ ID NO:12, and LCDR3 has the amino acid sequence shown in SEQ ID NO:14;

[0088] 7) HCDR1 has the amino acid sequence shown in SEQ ID NO: 1, HCDR2 has the amino acid sequence shown in SEQ ID NO: 3, HCDR3 has the amino acid sequence shown in SEQ ID NO: 4, LCDR1 has the amino acid sequence shown in SEQ ID NO: 11, LCDR2 has the amino acid sequence shown in SEQ ID NO: 12, and LCDR3 has the amino acid sequence shown in SEQ ID NO: 14;

[0089] 8) HCDR1 has the amino acid sequence shown in SEQ ID NO: 1, HCDR2 has the amino acid sequence shown in SEQ ID NO: 3, HCDR3 has the amino acid sequence shown in SEQ ID NO: 4, LCDR1 has the amino acid sequence shown in SEQ ID NO: 10, LCDR2 has the amino acid sequence shown in SEQ ID NO: 12, and LCDR3 has the amino acid sequence shown in SEQ ID NO: 14;

[0090] 9) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:56, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:63, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:74, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:86, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:94, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:103;

[0091] 10) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:57, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:64, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:75, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:87, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:95, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:104;

[0092] 11) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:56, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:63, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:74, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:86, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:94, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:103;

[0093] 12) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:57, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:64, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:75, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:87, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:95, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:104;

[0094] 13) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:65, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:76, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:96, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:105;

[0095] 14) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:59, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:66, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:77, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:89, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:97, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:106;

[0096] 15) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:60, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:67, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:78, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:90, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:98, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:107;

[0097] 16) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:69, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:80, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:109;

[0098] 17) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:70, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:81, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:109;

[0099] 18) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:65, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:82, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:101, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:110;

[0100] 19) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:62, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:71, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:83, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:92, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:102, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:111;

[0101] 20) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:72, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:84, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:93, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:112; or

[0102] 21) The heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:73, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:85, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:113.

[0103] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen-binding portion thereof, comprising a heavy chain variable region (VH), wherein the VH has any one of the amino acid sequences shown in SEQ ID NOs: 15-20, 114-120, 122-128.

[0104] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain variable region (VH), wherein VH has any one of the amino acid sequences shown in SEQ ID NOs: 16-20, 114-120, 122-128.

[0105] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain variable region (VH), wherein the VH has the amino acid sequence shown in SEQ ID NO: 18, 114-120, 122-128.

[0106] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen binding portion thereof, comprising a light chain variable region (VL), wherein the VL has any one of the amino acid sequences shown in SEQ ID NOs: 21-28, 129-135, 137-143.

[0107] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain variable region (VL), wherein VL has any one of the amino acid sequences shown in SEQ ID NOs: 22-28, 129-135, 137-143.

[0108] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain variable region (VL), wherein VL has any one of the amino acid sequences shown in SEQ ID NOs: 24, 25, 28, 129-135, 137-143.

[0109] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain variable region (VL), wherein VL has any one of the amino acid sequences shown in SEQ ID NOs: 25, 28, 129-135, 137-143.

[0110] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain variable region (VL), wherein the VL has the amino acid sequence shown in SEQ ID NO: 28, 129-135, 137-143.

[0111] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0112] 1) VH has the amino acid sequence shown in SEQ ID NO: 15, and VL has the amino acid sequence shown in SEQ ID NO: 21;

[0113] 2) VH has the amino acid sequence shown in SEQ ID NO: 16, and VL has the amino acid sequence shown in SEQ ID NO: 22;

[0114] 3) VH has the amino acid sequence shown in SEQ ID NO: 17, and VL has the amino acid sequence shown in SEQ ID NO: 23;

[0115] 4) VH has the amino acid sequence shown in SEQ ID NO: 18, and VL has the amino acid sequence shown in SEQ ID NO: 23;

[0116] 5) VH has the amino acid sequence shown in SEQ ID NO: 19, and VL has the amino acid sequence shown in SEQ ID NO: 23;

[0117] 6) VH has the amino acid sequence shown in SEQ ID NO: 18, and VL has the amino acid sequence shown in SEQ ID NO: 24;

[0118] 7) VH has the amino acid sequence shown in SEQ ID NO: 19, and VL has the amino acid sequence shown in SEQ ID NO: 24;

[0119] 8) VH has the amino acid sequence shown in SEQ ID NO: 20, and VL has the amino acid sequence shown in SEQ ID NO: 25;

[0120] 9) VH has the amino acid sequence shown in SEQ ID NO: 20, and VL has the amino acid sequence shown in SEQ ID NO: 26;

[0121] 10) VH has the amino acid sequence shown in SEQ ID NO: 20, and VL has the amino acid sequence shown in SEQ ID NO: 27;

[0122] 11) VH has the amino acid sequence shown in SEQ ID NO: 20, and VL has the amino acid sequence shown in SEQ ID NO: 28;

[0123] 12) VH has the amino acid sequence shown in SEQ ID NO: 18, and VL has the amino acid sequence shown in SEQ ID NO: 25;

[0124] 13) VH has the amino acid sequence shown in SEQ ID NO: 18, and VL has the amino acid sequence shown in SEQ ID NO: 28;

[0125] 14) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 114, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 129;

[0126] 15) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 115, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 130;

[0127] 16) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 116, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 131;

[0128] 17) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 117, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 132;

[0129] 18) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 118, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 133;

[0130] 19) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 119, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 134;

[0131] 20) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 120, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 135;

[0132] 21) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 122, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 137;

[0133] 22) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 123, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 138;

[0134] 23) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 124, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 139;

[0135] 24) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 125, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 140;

[0136] 25) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 126, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 141;

[0137] 26) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 127, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 142; or

[0138] 27) The heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 127, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 143.

[0139] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen-binding portion thereof, comprising a heavy chain constant region (CH), wherein the CH has any one of the amino acid sequences shown in SEQ ID NOs: 29-30.

[0140] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain constant region (CH), wherein the CH has the amino acid sequence shown in SEQ ID NO:30.

[0141] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain constant region (CL), wherein the CL has the amino acid sequence shown in SEQ ID NO:31.

[0142] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain constant region (CH) and a light chain constant region (CL), wherein:

[0143] 1) CH has the amino acid sequence shown in SEQ ID NO:29, and CL has the amino acid sequence shown in SEQ ID NO:31;

[0144] 2) CH has the amino acid sequence shown in SEQ ID NO:30, and CL has the amino acid sequence shown in SEQ ID NO:31;

[0145] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen binding portion thereof, comprising a heavy chain, wherein the heavy chain has any one of the amino acid sequences shown in SEQ ID NOs: 32-39, 144-150, 152-158.

[0146] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain, wherein the heavy chain has any one of the amino acid sequences shown in SEQ ID NOs: 34-39, 144-150, 152-158.

[0147] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain, wherein the heavy chain has the amino acid sequence shown in SEQ ID NO: 37, 144-150, 152-158.

[0148] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen binding portion thereof, comprising a light chain, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 40-47, 159-165, 167-173.

[0149] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 41-47, 159-165, 167-173.

[0150] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 43, 44 and 47, 159-165, 167-173.

[0151] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain, wherein the light chain has the amino acid sequence shown in any one of SEQ ID NOs: 44 and 47, 159-165, 167-173.

[0152] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a light chain, wherein the light chain has the amino acid sequence shown in SEQ ID NO: 47, 159-165, 167-173.

[0153] In certain aspects, the anti-CCR8 antibody or antigen binding protein comprises a heavy chain and a light chain, wherein:

[0154] 1) the heavy chain has the amino acid sequence shown in SEQ ID NO:32, and the light chain has the amino acid sequence shown in SEQ ID NO:40;

[0155] 2) the heavy chain has the amino acid sequence shown in SEQ ID NO:33, and the light chain has the amino acid sequence shown in SEQ ID NO:41;

[0156] 3) the heavy chain has the amino acid sequence shown in SEQ ID NO:34, and the light chain has the amino acid sequence shown in SEQ ID NO:42;

[0157] 4) the heavy chain has the amino acid sequence shown in SEQ ID NO:35, and the light chain has the amino acid sequence shown in SEQ ID NO:42;

[0158] 5) the heavy chain has the amino acid sequence shown in SEQ ID NO:36, and the light chain has the amino acid sequence shown in SEQ ID NO:42;

[0159] 6) the heavy chain has the amino acid sequence shown in SEQ ID NO:35, and the light chain has the amino acid sequence shown in SEQ ID NO:43;

[0160] 7) the heavy chain has the amino acid sequence shown in SEQ ID NO:36, and the light chain has the amino acid sequence shown in SEQ ID NO:43;

[0161] 8) the heavy chain has the amino acid sequence shown in SEQ ID NO:38, and the light chain has the amino acid sequence shown in SEQ ID NO:41;

[0162] 9) the heavy chain has the amino acid sequence shown in SEQ ID NO:39, and the light chain has the amino acid sequence shown in SEQ ID NO:44;

[0163] 10) the heavy chain has the amino acid sequence shown in SEQ ID NO:39, and the light chain has the amino acid sequence shown in SEQ ID NO:45;

[0164] 11) the heavy chain has the amino acid sequence shown in SEQ ID NO:39, and the light chain has the amino acid sequence shown in SEQ ID NO:46;

[0165] 12) the heavy chain has the amino acid sequence shown in SEQ ID NO:39, and the light chain has the amino acid sequence shown in SEQ ID NO:47;

[0166] 13) the heavy chain has the amino acid sequence shown in SEQ ID NO:37, and the light chain has the amino acid sequence shown in SEQ ID NO:43;

[0167] 14) the heavy chain has the amino acid sequence shown in SEQ ID NO:37, and the light chain has the amino acid sequence shown in SEQ ID NO:44;

[0168] 15) the heavy chain has the amino acid sequence shown in SEQ ID NO:37, and the light chain has the amino acid sequence shown in SEQ ID NO:47;

[0169] 16) the heavy chain has the amino acid sequence shown in SEQ ID NO: 144, and the light chain has the amino acid sequence shown in SEQ ID NO: 159;

[0170] 17) the heavy chain has the amino acid sequence shown in SEQ ID NO: 145, and the light chain has the amino acid sequence shown in SEQ ID NO: 160;

[0171] 18) the heavy chain has the amino acid sequence shown in SEQ ID NO: 146, and the light chain has the amino acid sequence shown in SEQ ID NO: 161;

[0172] 19) the heavy chain has the amino acid sequence shown in SEQ ID NO: 147, and the light chain has the amino acid sequence shown in SEQ ID NO: 162;

[0173] 20) the heavy chain has the amino acid sequence shown in SEQ ID NO: 148, and the light chain has the amino acid sequence shown in SEQ ID NO: 163;

[0174] 21) the heavy chain has the amino acid sequence shown in SEQ ID NO: 149, and the light chain has the amino acid sequence shown in SEQ ID NO: 164;

[0175] 22) the heavy chain has the amino acid sequence shown in SEQ ID NO: 150, and the light chain has the amino acid sequence shown in SEQ ID NO: 165;

[0176] 23) the heavy chain has the amino acid sequence shown in SEQ ID NO: 152, and the light chain has the amino acid sequence shown in SEQ ID NO: 167;

[0177] 24) the heavy chain has the amino acid sequence shown in SEQ ID NO: 153, and the light chain has the amino acid sequence shown in SEQ ID NO: 168;

[0178] 25) the heavy chain has the amino acid sequence shown in SEQ ID NO: 154, and the light chain has the amino acid sequence shown in SEQ ID NO: 169;

[0179] 26) the heavy chain has the amino acid sequence shown in SEQ ID NO: 155, and the light chain has the amino acid sequence shown in SEQ ID NO: 170;

[0180] 27) the heavy chain has the amino acid sequence shown in SEQ ID NO: 156, and the light chain has the amino acid sequence shown in SEQ ID NO: 171;

[0181] 28) the heavy chain has the amino acid sequence shown in SEQ ID NO: 157, and the light chain has the amino acid sequence shown in SEQ ID NO: 172; or

[0182] 29) The heavy chain has the amino acid sequence shown in SEQ ID NO: 158, and the light chain has the amino acid sequence shown in SEQ ID NO: 173.

[0183] Certain aspects of the present disclosure relate to defucosylated anti-CCR8 antibodies, or antigen-binding portions thereof.

[0184] Certain aspects of the present disclosure relate to anti-CCR8 antibodies, or antigen-binding portions thereof, which induce antibody-dependent cellular cytotoxicity (ADCC) in a subject upon administration of the anti-CCR8 antibodies, or antigen-binding portions thereof.

[0185] Certain aspects of the present disclosure relate to anti-CCR8 antibodies, or antigen-binding portions thereof, that are capable of inducing activation of NK cells.

[0186] In certain aspects, the present disclosure relates to an anti-CCR8 antibody, or antigen-binding portion thereof, that is capable of inducing NK cell-mediated killing of tumor-infiltrating Treg cells in a subject.

[0187] In certain aspects, the disclosure relates to anti-CCR8 antibodies, or antigen-binding portions thereof, that induce internalization of CCR8 by tumor-infiltrating Treg cells.

[0188] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or an antigen binding portion thereof, that is a human, humanized, or chimeric antibody, or an antigen binding portion thereof.

[0189] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or antigen-binding portion thereof, that is an IgG1, IgG2, IgG3, or IgG4 isotype antibody, or antigen-binding fragment thereof.

[0190] Certain aspects of the present disclosure relate to an anti-CCR8 antibody, or an antigen-binding portion thereof, which is a monoclonal antibody, a bispecific antibody, a bispecific T cell engager (BiTE), a multispecific antibody, a biparatopic antibody, an immunoconjugate, an antibody drug conjugate, a chimeric antigen receptor (CAR), a T cell receptor (TCR), or any combination thereof.

[0191] Certain aspects of the present disclosure relate to anti-CCR8 antibodies, or antigen-binding portions thereof, that target the extracellular domain of human CCR8.

[0192] In certain aspects, the disclosure relates to anti-CCR8 antibodies, or antigen binding portions thereof, that bind to cells expressing CCR8 on their surface.

[0193] In certain aspects, the disclosure relates to anti-CCR8 antibodies, or antigen-binding portions thereof, that specifically bind to one or more amino acids within the N-terminal extracellular domain of CCR8.

[0194] In certain aspects, the disclosure relates to anti-CCR8 antibodies, or antigen binding portions thereof, that bind to cells expressing CCR8 on their surface, wherein the cells are regulatory T cells (Tregs).

[0195] In certain aspects, the disclosure relates to anti-CCR8 antibodies, or antigen binding portions thereof, that bind to cells expressing CCR8 on their surface, wherein the cells are tumor infiltrating Tregs.

[0196] Certain aspects of the present disclosure relate to anti-CCR8 antibodies, or antigen-binding portions thereof, that compete for binding with CCL1.

[0197] Certain aspects of the present disclosure relate to a pharmaceutical composition comprising an anti-CCR8 antibody, or antigen-binding portion thereof, according to the present disclosure and a pharmaceutically acceptable carrier.

[0198] Certain aspects of the present disclosure relate to methods of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject an anti-CCR8 antibody or antigen-binding portion according to the present disclosure or a pharmaceutical composition according to the present disclosure.

[0199] In certain aspects, the disease or disorder comprises a tumor.

[0200] In certain aspects, the present disclosure relates to methods of treating a tumor in a subject in need thereof, wherein the tumor comprises Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic promyelocytic myelomonocytic monocytic erythroleukemia, chronic leukemia, chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt's lymphoma and marginal zone B-cell lymphoma, polycythemia vera lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's macroglobulinemia, macroglobulinemia), heavy chain disease, solid tumors, sarcomas and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon sarcoma, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma (HCC), liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular tumors, lung cancer, small cell lung cancer, non-small cell lung cancer, bladder tumors, epithelial cancer, glioma, astrocytoma, Medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal carcinoma, esophageal cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, stomach cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell lung cancer, large cell lung cancer), melanoma, neuroblastoma; oral cancer (e.g., lip cancer, tongue cancer, mouth cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; respiratory system cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and urinary system cancer, and any combination thereof.

[0201] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, wherein the tumor is refractory or recurrent.

[0202] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, wherein the tumor is advanced, locally advanced, or metastatic.

[0203] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, the method further comprising administering an additional anti-cancer agent and / or method.

[0204] Certain aspects of the present disclosure relate to the use of an anti-CCR8 antibody, or antigen-binding portion thereof, according to the present disclosure in the manufacture of a medicament for treating a disease or disorder.

[0205] In certain aspects, the disease or disorder comprises a tumor.

[0206] In certain aspects, the present disclosure relates to a method of treating a tumor in a subject in need thereof, wherein the tumor includes Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic promyelocytic myelomonocytic monocytic erythroleukemia, chronic leukemia, chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt's lymphoma and marginal zone B-cell lymphoma, polycythemia vera lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Washington University in St. Louis, MO Macroglobulinemia, heavy chain disease, solid tumors, sarcomas and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon sarcoma, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, liver HCC, liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular tumors, lung cancer, small cell lung cancer, non-small cell lung cancer, bladder tumors, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal carcinoma, esophageal cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, stomach cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell lung cancer, large cell lung cancer), melanoma, neuroblastoma; oral cancer (e.g., lip cancer, tongue cancer, mouth cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; respiratory system cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and urinary system cancer, and any combination thereof.

[0207] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, wherein the tumor is refractory or recurrent.

[0208] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, wherein the tumor is advanced, locally advanced, or metastatic.

[0209] In certain aspects, the disclosure relates to methods of treating a tumor in a subject in need thereof, the method further comprising administering an additional anti-cancer agent and / or method.

[0210] Example

[0211] Example 1: Generation of various anti-CCR8 antibodies

[0212] 1.1 Hybridoma

[0213] To generate antibodies against CCR8, Sprague-Dawley rats were immunized with CCR8 overexpressing cells using a variety of primary immunity-boosting strategies. To monitor immune responses, titrated serum was screened for binding to a variety of CCR8 overexpressing cell lines by FACS, typically 4-6 weeks after immunization. Animals with sufficient titers of anti-CCR8 IgG were selected for final immunization.

[0214] In order to produce hybridomas, cells from lymphoid organs (such as spleen and lymph nodes) are collected and separated and fused with SP2 / 0 myeloma cells. The obtained cells are plated in a flat 96-well plate supplemented with culture medium E (StemCell Technologies) of HAT (Sigma H0262-10VL) to select hybridomas. After one week of cultivation, hybridoma supernatants are collected and the cell lines overexpressing CCR8 are screened by flow cytometry. Subclones subsequently secrete hybridomas of anti-CCR8 antibodies and further screen them to identify anti-CCR8 monoclonal hybridomas.

[0215] 1.2 Generation of expression vector

[0216] To obtain the sequences of the HC and LC variable domains, mRNA from hybridomas was purified using the Dynabeads mRNA Direct Micro Purification Kit (ThermoFisher #61021), and cDNA was synthesized and amplified using template-switched oligonucleotides (Pinto et al., Anal. Biochem. 397, 2010). The variable domain fragments of the heavy and light chains were subsequently amplified, purified, and sequenced.

[0217] To generate expression vectors, variable regions of heavy and light chain DNA sequences were synthesized and subcloned in-frame with either human IgG1 kappa constant region light chain or human IgG1 constant heavy chain with SDIE mutation previously inserted into pCI vector (Promega #E1841).

[0218] 1.3 Humanization of antibodies

[0219] To humanize the antibody, the CDR residues of the light and heavy chains were grafted onto a human framework with high homology. To restore affinity, some mutations were designed by changing the human framework residues to their murine counterparts at specific positions.

[0220] 1.4 Antibody Production

[0221] Antibodies were produced by co-transfection of plasmids into Expi293F cells (Thermo Fisher Scientific A14527). Specifically, 30 μg of total DNA was diluted into Opti-MEM medium (Life Technologies) and then incubated with Expifectamine 293 transfection reagent (Gibco #A14525) in the same medium for 20 min. The mixture was then added to 30 ml of Expi293F cells grown in suspension at a density of 2.5 million cells / ml at 37°C and 8% CO2.

[0222] After transfection, cells were grown in suspension for about six days and then harvested by centrifugation at 7,000 rpm at 4 ° C for 20 minutes. The supernatant was filtered through a 0.22 μm filter and affinity purified using protein A resin. Protein was eluted by elution buffer (1M glycine pH 3.0, 10% glycerol) and neutralized to pH 6.0 with 1M Tris (pH 7.5). Size exclusion chromatography (SEC) was performed on AKTA to refine the product, and the antibody concentration was measured at 280 nm using a micro-spectrophotometer (NanoDrop). Antibodies were analyzed by electrophoresis on 4%-20% Tris-glycine gel (Bio-Rad) under reduced (R) and non-reduced (NR) conditions.

[0223] The antibodies listed in Table 1 below were obtained by conventional experimental procedures for antibody humanization well known in the art, using 433H (BD company #566379) or B1 (Biolegend company #360602) as parent antibodies. These procedures include, but are not limited to, changing IgG subclass, re-humanization, changing signal peptides, amino acid point mutations, and / or a combination of several of these methods. For example, chimeric antibodies (ChH2 and ChB1) were constructed by transplanting the VL and VH regions of mouse antibodies 433H or B1 onto human IgG1 constant regions.

[0224] Defucosylated antibodies were produced by transfecting the heavy and light chain plasmids of the target antibody into CHO cells lacking mammalian α1,6-fucosyltransferase (FUT8) (Sino Biologics). After 7 days, the culture medium was harvested and the antibody was purified using a protein A column. The purified antibodies were analyzed by SDS-PAGE and SEC-HPLC. Thus, three defucosylated antibodies were obtained: ChH2-AF, ABC024-H2G.AF and ChB1-AF.

[0225] 1.5 mAb analysis test (HPLC)

[0226] The purity and monomer content of the final purified protein were determined by high throughput analysis on HPLC. Size exclusion chromatography (SEC) was performed on an Infinity 1260II Bio-Inert Agilent HPLC system using Advancebio SEC 300A 4.6x 300mm, 2.7 μm (Agilent PL1580-5301). Mobile phase PBS was used to inject under isocratic elution conditions and absorbance was detected at 280nm. Quantification was performed based on the relative area of ​​the detected peak.

[0227] The subject antibody can be substantially pure, e.g., at least about 80% to 85% pure, at least about 85% to 90% pure, at least about 90% to 95% pure, or 98% to 99% or more pure, e.g., free of contaminants such as cellular debris, macromolecules other than the subject antibody, etc.

[0228] Tables 1 and 2 show the full-length amino acid sequences of these antibodies.

[0229] Table 1. Amino acid sequences of antibodies of the present disclosure

[0230]

[0231]

[0232] Table 2. Amino acid sequences of antibodies of the present disclosure

[0233] ID Antibody Heavy chain Light chain 1 ABC519 SEQ ID NO:144 SEQ ID NO:159 2 ABC523 SEQ ID NO:145 SEQ ID NO:160 3 ABC527 SEQ ID NO:146 SEQ ID NO:161 4 ABC531 SEQ ID NO:147 SEQ ID NO:162 5 ABC814 SEQ ID NO:148 SEQ ID NO:163 6 ABC815 SEQ ID NO:149 SEQ ID NO:164 7 ABC816 SEQ ID NO:150 SEQ ID NO:165 8 ABC818 SEQ ID NO:152 SEQ ID NO:167 9 ABC819 SEQ ID NO:153 SEQ ID NO:168 10 ABC820 SEQ ID NO:154 SEQ ID NO:169 11 ABC821 SEQ ID NO:155 SEQ ID NO:170 12 ABC822 SEQ ID NO:156 SEQ ID NO:171 13 ABC823 SEQ ID NO:157 SEQ ID NO:172 14 ABC1183 SEQ ID NO:158 SEQ ID NO:173

[0234] Example 2: Cell Binding or Affinity of Anti-CCR8 Antibodies

[0235] 2.1 Generation of stable CCR8-overexpressing cell lines

[0236] In order to generate cell lines for animal immunization and antibody characterization, a cell line overexpressing CCR8 was developed. The nucleotide sequence (SEQ ID NO: 50) encoding human CCR8 (Uniprot # P51685, SEQ ID NO: 51) was cloned into pIREShygro3gGFP, which was modified from pIREShyg3 (Takara # 631620) by fusing the eGFP cDNA with the nucleotide sequence of SEQ ID NO: 48 to the 3' hygromycin B phosphotransferase gene in pIREShyg3 using a linker with the nucleotide sequence of SEQ ID NO: 49. The resulting plasmid was then transfected into HEK293T cells (ATCC CRL-3216) and C6 cells (ATCC CCL-107) using lipofectamine 3000 (Thermo Fisher Scientific, L3000001). Three days later, cells were treated with hygromycin B (Millipore Sigma) for two weeks to select stable cell lines with high CCR8 cell surface expression.

[0237] 2.2 Cell-based binding assay of human CCR8 on cell lines

[0238] The binding specificity of anti-CCR8 antibody (mAb) was tested by FACS using several cell lines. Using TALL-1 (DSMZ # ACC 521) and Hut78 cell lines (ATCC # TIB-161), both of which carry endogenous human CCR8 on their cell surfaces, HEK293T cells overexpressing CCR8 were also used. The binding titration of anti-CCR8 antibody and CCR8 transfectant was carried out by serial dilution of the antibody. The diluted antibody (50 μl) in flow cytometry buffer (PBS, 0.5% BSA, 1 mM EDTA) was incubated on ice for 30 min with cells (50 μl) at a concentration of 2 million cells / ml. After washing twice with flow cytometry buffer, the antibody bound was detected on ice for 20 minutes using F (ab') 2 fragment goat anti-human IgG (Jackson ImmunoResearch Laboratories, Inc. (Jackson ImmunoResearch) # 709-606-149) (1: 500 dilution) labeled with Alexa647. After washing twice with flow cytometry buffer, fluorescence was measured on a Cytek Aurora cytometer. Data were analyzed using GraphPad Prism 8.0 software to determine EC50 values.

[0239] 2.3 Cell-based binding assay of human PBMC-derived activated Tregs

[0240] The binding specificity of anti-CCR8 antibodies was tested by FACS using activated Treg derived from human PBMC. In short, Treg was amplified and activated according to the manufacturer's protocol (130-095-345, Miltenyi). In brief, the human PBMC-derived Treg (SailyBio) of the amplification frozen for 7 days was thawed and left to stand overnight. Then Treg was restimulated with CD3 / CD28 MACSiBeads in the presence of rhIL-2 for 24 hours. CCR8 expression on these activated Treg was confirmed using anti-human CCR8 (clone 433H, BD) or anti-human CCR8 (clone L263G8, Biolegend). Then, these activated Treg were plated in 96-well plates with 20K / wells, centrifuged and the supernatant was removed. The cell pellet was then resuspended and incubated with 100 μl of various concentrations of mAb or human IgG1 isotype control antibody (biolegend company #403501) at 37°C for 30 min. The final concentration of the antibody is shown on the X-axis of each figure. The cells were washed three times with FACS buffer (PBS containing 0.5 mg / ml BSA and 1 mM EDTA). PE-labeled goat anti-human IgG (Invitrogen) diluted in FACS buffer was added to the cell pellet as a secondary antibody at a concentration of 3 μg / ml, and the samples were incubated at 4°C for 30 min. The samples were washed three times with FACS buffer and analyzed using a Cytek Aurora cytometer. The data were analyzed using GraphPad Prism8.0 software to determine the EC50 value.

[0241] Figure 1-4 Table 3 shows that all antibodies listed in Table 1 can bind to human CCR8 on the cell surface. In particular, ABC137-H2.hu12.IgG1.SDIE and ABC138-H2.hu13.IgG1.SDIE showed higher affinity for cells expressing CCR8 than their parental clones ABC025.H2G.SDIE. In addition, ABC138-H2.hu13.IgG1.SDIE showed the highest binding ability to activated Treg ( Figure 4 A negative control antibody (ABC089, having a heavy chain amino acid sequence as shown in SEQ ID NO: 178 and a light chain sequence as shown in SEQ ID NO: 179) was derived from PDB 2EIZ.

[0242] Table 3 shows the affinity of anti-CCR8 mAb binding to activated Treg derived from human PBMC, demonstrating that the antibodies disclosed herein are much more effective than prior art ABC084. AF (disclosed in PCT / EP 2021 / 067504) having a heavy chain as shown in SEQ ID NO: 52 and a light chain amino acid sequence as shown in SEQ ID NO: 53. In particular, ABC048-B1.hu11.IgG1.SDIE and ABC138-H2.hu13.IgG1.SDIE are even more effective than prior art ABC078.AF (also known as BMS-986340 of BMS, disclosed in PCT / US2021 / 023430), which has a heavy chain as shown in SEQ ID NO: 54 and a light chain amino acid sequence as shown in SEQ ID NO: 55, and has entered Phase I / II trials.

[0243] Table 3. EC50 of anti-CCR8 antibody binding to human PBMC-derived activated Tregs

[0244] mAbs EC50(nM) ABC025.H2G.SDIE 1.776 ABC048.B1.hu11.IgG1.SDIE 0.5214 ABC0138.H2.Hu13.SDIE 0.5790 ABC078.AF 0.7286 ABC084.AF 33.77

[0245] also, Figure 5-6 As shown in Tables 4-6, all antibodies in Table 2 have the ability to bind to human CCR8 on the cell surface. Among them, ABC822 and its humanized form ABC1183 show high cross-reactivity to human and cynomolgus monkey CCR8 and endogenous CCR8 on the Hut78 cell line. Prior art antibody ABC026 (disclosed in PCT / US2021 / 017268) has a heavy chain as shown in SEQ ID NO: 174 and a light chain amino acid sequence as shown in SEQ ID NO: 175. Antibody ABC080.chH2.SDIE has a heavy chain as shown in SEQ ID NO: 176 and a light chain amino acid sequence as shown in SEQ ID NO: 177.

[0246] Table 4. EC50 of antibody binding to CCR8 on cell lines

[0247]

[0248]

[0249] "NA" means no data

[0250] Table 5. EC50 for antibody binding to cell lines expressing CCR8.

[0251] EC50(nM) Hut78 293T-hCCR8 293T-cCCR8 293T-mCCR8 ABC822 2.917 0.2689 0.4637 No combination ABC1183 4.121 0.3727 0.4448 No combination

[0252] In particular, Figure 6Table 6 shows the titration of anti-CCR8 mAbs ABC138-H2.hu13.IgG1.SDIE, ABC025-H2G.SDIE and ABC080.chH2.SDIE on human PBMC-derived activated Treg cells. ABC138-H2.hu13.IgG1.SDIE was generated by mutagenesis in the light chain CDR of ABC025.H2G.SDIE (which is a humanized clone of ABC080.chH2.SDIE). The binding of anti-CCR8 monoclonal antibodies was then tested using activated Tregs derived from human PBMCs, as shown in Table 7. Figure 6 As described in Table 6, the results showed that ABC138-H2.hu13.IgG1.SDIE exhibited higher affinity than both chimeric ABC080.chH2.SDIE and humanized ABC025.H2G.SDIE.

[0253] Table 6. EC50 of anti-CCR8 mAb binding to human PBMC-derived activated Treg cells

[0254] Anti-CCR8 mAb EC50(nM) ABC138-H2.hu13.IgG1.SDIE 0.1051 ABC025.H2G.SDIE 0.3799 ABC080.chH2.SDIE 0.3799

[0255] Example 3: Competition assay between the anti-CCR8 antibodies of the present disclosure and CCL1

[0256] CC chemokine (CCL1) is a potent chemoattractant for leukocytes via its interaction with CCR8. To test the effectiveness of anti-human CCR8 antibodies in blocking this interaction, calcium flux assays were performed on the CHEM1-CCR8 cell line (Eurofins #HTS013RTA), which expresses high levels of human CCR8 on the cell surface and contains the promiscuous G protein Gα15 required to couple the receptor to the calcium signaling pathway. CHEM1-CCR8 cells were cultured overnight in 96-well plates at a density of 70K / well and labeled with Calbryte 520AM (Biomol #ABD-20650). After discarding the labeling buffer, 60 μl of anti-CCR8 mAb diluted in HBSS buffer was added to the cells and incubated at room temperature for 15 minutes. Then, 2,000 nM (Biolegend #582702) of human CCL1 protein (20 μl) was injected into each well via the Flexstation-flex3 system to reach a final concentration of 500 nM, and the calcium flux signal was monitored for 90 seconds using specific settings: fluorescence bottom reading mode, excitation = 490 nm, emission = 525 nm, cutoff = 515 nm, time = 90 s, interval = 2.1 s, readings = 43.

[0257] Figure 7-Figure 8Anti-human CCR8 ChH2-AF was shown to block human CCL1 to human CCR8 signaling, while anti-human CCR8 ChB1-AF had limited effects on CCL1-CCR8 signaling. Studies have shown that CCL1 binding to CCR8 can further enhance the suppressive activity of Tregs in in vitro assays and inhibit autoimmune inflammatory responses in mouse models. Therefore, anti-CCR8 antibodies that inhibit CCL1 binding to CCR8 may have better anti-tumor efficacy, and disrupting CCL1-CCR8 interactions may further increase the anti-tumor potency of anti-CCR8 mAbs with enhanced effector functions.

[0258] Example 4: ADCC Potential of Anti-CCR8 Antibodies

[0259] To evaluate the ADCC ability of anti-CCR8 mAbs, primary human NK cells were used as effector cells, while TALL-1 (DSMZ#ACC 521) or Hut78 (ATCC#TIB-161) were used as target cells ( Figure 9-11 NK cells were enriched from PBMC using a negative selection kit (Biolegend #480053) and confirmed to be CD3-CD56+ with a purity of Fig. 9 74% of Fig.10 88.8% and Fig.11 Purified NK cells had FcγRIIIA 158V / F or V / V genotype.

[0260] Target cells were labeled with CellTracer violet (Thermo #C34557) and various antibodies were added to the cell mixture. Fig. 9 In the NK cell culture, 133K / well NK cells and 16,625 / well TALL-1 target cells were used, with a ratio of 8:1. Fig.10 In the experiment, 160K NK cells / well and 20K Hut78 target cells / well were used, with a ratio of 8:1. Fig.11 In the experiment, 128K / well NK cells and 20K / well Hut78 target cells were used at a ratio of 6.4:1. NK cells and target cells were co-cultured with different antibodies in U-bottom 96-well plates at 37°C for 4 hours, and cell killing was assessed using Live / Dead FixableNear IR stain (Thermo #L34994) at a dilution of 1:1000 and quantified by Cytek Aurora cytometer using Cytek default settings. EC50 was calculated using PRISM using the Sigmoidal 4-parameter model.

[0261] Fig. 9As shown in Table 7, primary NK cells can effectively mediate ADCC of CCR8 expressing cells when treated with anti-CCR8 mAb ChH2-AF or ChB1-AF.

[0262] Table 7. ADCC killing of TALL-1 cells mediated by defucosylated anti-CCR8 mAb

[0263] Anti-CCR8 mAb Median EC50 (pM) ChH2-AF 2.546 ChB1-AF 9.799

[0264] The ADCC potential of anti-CCR8 mAbs, such as AB024-H2G.AF, ABC025.H2G.SDIE and ABC024-H2G, is as follows: Fig.10 Compared to ABC024-H2G with wild-type Fc, Fc engineering by defucosylation (e.g. AB024-H2G.AF) or SDIE mutation (e.g. ABC025.H2G.SDIE) enhanced killing of target cells in ADCC assays.

[0265] Fig.11 The dose-dependent killing potency of ABC024-H2G.AF and ABC025-H2G.SDIE was compared in an ADCC assay. These two molecules, which share variable domains and employ different effector function enhancement technologies, exhibited very comparable ADCC potential in this assay.

[0266] 4.2 ADCC assay against Jurkat-h / cyno-CCR8 or Hut78 cells

[0267] To generate Jurkat cell lines overexpressing human or cynomolgus monkey CCR8, the target gene was cloned into a lentiviral transfer vector (VPK-206, Cell Biolabs). Lentiviral particles were produced in 293T cells by transient transfection of a lentiviral transfer vector carrying CCR8 and the ViraSafe lentiviral packaging system (Cell Biolabs #VPK-206). Jurkat cells were transduced with viral particles in the presence of 10 μg / ml polybrene and then treated with 1 μg / ml puromycin for 14 days. CCR8 overexpression on Jurkat cells was confirmed by flow cytometry.

[0268] For ADCC assay, PBMC effectors from healthy donors were thawed and placed overnight in ADCC medium (RPMI 1640 with glutamine, 10% low IgG heat-inactivated FBS, 1% Glutamix, 1% non-essential amino acids, 1% sodium pyruvate). The cells were then collected and washed with ADCC wash buffer (1% BSA in DPBS) to be used as effector cells.

[0269] Jurkat-hCCR8, Jurkat-cyno-CCR8 and Hut78 cells were labeled with CellTraceViolet stain (Invitrogen, catalog number C34557) diluted 10,000 times for 20 min, and then washed twice with ADCC wash buffer. The labeled cells were resuspended in ADCC medium and plated in 96-well U-bottom plates at 30,000 cells / well 50 μl / well as target cells.

[0270] After incubation of target cells with 50 μl of serially diluted antibodies at room temperature for 20 minutes, effector cells (effector cell: target cell (E:T) ratio of 8) were added to the plate. After 18 hours of treatment, the cells were stained with Live / Dead FixableNear IR (1:1000, Invitrogen, catalog number L34976) on ice for 20 minutes. Cell killing was analyzed using a Cytek Aurora cytometer and data were analyzed using GraphPad Prism 9.0 software to determine EC50.

[0271] ABC138 (ABC138-H2.hu13.IgG1.SDIE) and ABC078.AF showed limited ADCC activity against Jurkat-cyno-CCR8, consistent with their minimal cross-reactivity to cyno CCR8 ( Figure 12-14 and Tables 8-10). In contrast, ABC822 and its humanized mAb ABC1183 showed potent activity in killing Jurkat-cyno-CCR8 and Jurkat-hCCR8 cells. The negative control antibody (ABC089, having a heavy chain amino acid sequence as shown in SEQ ID NO: 178 and a light chain sequence as shown in SEQ ID NO: 179) was derived from PDB 2EIZ. However, ABC138-H2.hu13.IgG1.SDIE showed the highest activity in killing Hut78 cells expressing endogenous CCR8.

[0272] Table 8. ADCC killing of Jurkat-cynoCCR8 cells mediated by anti-CCR8 mAbs

[0273] Anti-CCR8 mAb IC50(pM) ABC138-H2.hu13.IgG1.SDIE 62.90 ABC078.AF 345.8 ABC1183 6.150 ABC822 7.190

[0274] Table 9. ADCC killing of Jurkat-hCCR8 cells mediated by anti-CCR8 mAbs

[0275] Anti-CCR8 mAb IC50(pM) ABC138-H2.hu13.IgG1.SDIE 2.088 ABC078.AF 2.788 ABC1183 1.796 ABC822 9.677

[0276] Table 10. ADCC killing of Hut78 cells mediated by anti-CCR8 mAb

[0277] Anti-CCR8 mAb IC50(pM) ABC138-H2.hu13.IgG1.SDIE 0.6519 ABC078.AF 0.6367 ABC1183 7.945 ABC822 6.306

Claims

1. An anti-CCR8 antibody or an antigen-binding portion thereof, comprising a heavy chain CDR1 having an amino acid sequence as shown in any one of SEQ ID NOs: 1-2, 56-60, and 62, a heavy chain CDR2 having an amino acid sequence as shown in any one of SEQ ID NOs: 3, 63-67, and 69-73, a heavy chain CDR3 having an amino acid sequence as shown in any one of SEQ ID NOs: 4-5, 74-78, and 80-85, a light chain CDR1 having an amino acid sequence as shown in any one of SEQ ID NOs: 6-11, 86-90, and 92-93, a light chain CDR2 having an amino acid sequence as shown in any one of SEQ ID NOs: 12, 94-98, and 100-102, and a light chain CDR3 having an amino acid sequence as shown in any one of SEQ ID NOs: 13-14, 103-107, and 109-113.

2. The anti-CCR8 antibody or antigen-binding portion thereof according to claim 1, wherein: 1) the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 4, 74-78, 80-85; and 2) The light chain CDR1 has any one of the amino acid sequences shown in SEQ ID NO: 7, 10-11, 86-90, 92-93.

3. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 2, wherein: 1) the light chain CDR1 has any one of the amino acid sequences shown in SEQ ID NOs: 10-11, 86-90, 92-93; and 2) The light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 14, 103-107, 109-113. 4 . The anti-CCR8 antibody or antigen-binding portion thereof according to claim 1 , wherein the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 10, 86-90, 92-93.

5. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 4, wherein: 1) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 1, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 4, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 6, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 13; 2) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:2, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:5, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:11, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:14; 3) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 1, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 4, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 7, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 13; 4) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:2, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:5, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:8, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:14; 5) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:2, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:5, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:9, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:14; 6) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:2, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:5, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:10, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:14; 7) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 1, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 4, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 11, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 14; 8) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 1, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 3, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 4, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 10, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 12, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 14; 9) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 56, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 63, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 74, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 86, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 94, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 103; 10) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 57, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 64, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 75, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 87, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 95, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 104; 11) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 56, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 63, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 74, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 86, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 94, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 103; 12) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 57, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 64, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 75, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 87, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 95, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 104; 13) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 65, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 76, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 96, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 105; 14) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 59, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 66, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 77, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 89, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 97, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 106; 15) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 60, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 67, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 78, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 90, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 98, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 107; 16) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 69, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 80, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 109; 17) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 70, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 81, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 109; 18) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 65, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 82, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 101, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 110; 19) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO: 62, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO: 71, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO: 83, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO: 92, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO: 102, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO: 111; 20) the heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:72, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:84, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:93, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:112; or 21) The heavy chain CDR1 has the amino acid sequence shown in SEQ ID NO:58, the heavy chain CDR2 has the amino acid sequence shown in SEQ ID NO:73, the heavy chain CDR3 has the amino acid sequence shown in SEQ ID NO:85, the light chain CDR1 has the amino acid sequence shown in SEQ ID NO:88, the light chain CDR2 has the amino acid sequence shown in SEQ ID NO:100, and the light chain CDR3 has the amino acid sequence shown in SEQ ID NO:

113.

6. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 5, comprising a heavy chain variable region, wherein the heavy chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 15-20, 114-120, 122-128.

7. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 6, wherein the heavy chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 16-20, 114-120, 122-128.

8. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 7, wherein the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 18, 114-120, 122-128.

9. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 8, comprising a light chain variable region; wherein the light chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 21-28, 129-135, 137-143.

10. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 9, wherein the light chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 22-28, 129-135, 137-143.

11. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 10, wherein the light chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 24, 25, 28, 129-135, 137-143.

12. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 11, wherein the light chain variable region has any one of the amino acid sequences shown in SEQ ID NOs: 25, 28, 129-135, 137-143.

13. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 12, wherein the light chain variable region has the amino acid sequence shown in SEQ ID NO: 28, 129-135, 137-143.

14. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 13, wherein: 1) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 15, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 21; 2) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 16, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 22; 3) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 17, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 23; 4) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 18, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 23; 5) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 19, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 23; 6) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 18, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 24; 7) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 19, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 24; 8) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 25; 9) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 26; 10) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 27; 11) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 28; 12) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 18, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 25; 13) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 18, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 28; 14) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 114, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 129; 15) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 115, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 130; 16) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 116, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 131; 17) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 117, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 132; 18) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 118, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 133; 19) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 119, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 134; 20) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 120, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 135; 21) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 122, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 137; 22) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 123, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 138; 23) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 124, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 139; 24) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 125, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 140; 25) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 126, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 141; 26) the heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 127, and the light chain variable region has the amino acid sequence shown in SEQ ID NO: 142; or 27) The heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 127, and the light chain variable region has the amino acid sequence shown in SEQ ID NO:

143.

15. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 14, comprising a heavy chain constant region, wherein the heavy chain constant region has any one of the amino acid sequences shown in SEQ ID NOs: 29-30.

16. The anti-CCR8 antibody, or antigen binding portion thereof, according to any one of claims 1 to 15, wherein the heavy chain constant region has the amino acid sequence shown in SEQ ID NO:

30.

17. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 16, comprising a light chain constant region, wherein the light chain constant region has the amino acid sequence shown in SEQ ID NO:

31.

18. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1 to 17, wherein: 1) the heavy chain constant region has the amino acid sequence shown in SEQ ID NO: 29, and the light chain constant region has the amino acid sequence shown in SEQ ID NO: 31; 2) The heavy chain constant region has the amino acid sequence shown in SEQ ID NO:30, and the light chain constant region has the amino acid sequence shown in SEQ ID NO:

31.

19. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 18, comprising a heavy chain, wherein the heavy chain has any one of the amino acid sequences shown in SEQ ID NOs: 32-39, 144-150, 152-158.

20. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 19, wherein the heavy chain has any one of the amino acid sequences shown in SEQ ID NOs: 34-39, 144-150, 152-158.

21. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 20, wherein the heavy chain has the amino acid sequence shown in SEQ ID NO: 37, 144-150, 152-158.

22. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 21, comprising a light chain, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 40-47, 159-165, 167-173.

23. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 22, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 41-47, 159-165, 167-173.

24. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 23, wherein the light chain has any one of the amino acid sequences shown in SEQ ID NOs: 43, 44 and 47, 159-165, 167-173.

25. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 24, wherein the light chain has the amino acid sequence shown in any one of SEQ ID NOs: 44 and 47, 159-165, 167-173.

26. The anti-CCR8 antibody, or antigen binding portion thereof, according to any one of claims 1 to 25, wherein the light chain has the amino acid sequence shown in SEQ ID NO: 47, 159-165, 167-173.

27. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 26, wherein: 1) the heavy chain has the amino acid sequence shown in SEQ ID NO: 32, and the light chain has the amino acid sequence shown in SEQ ID NO: 40; 2) the heavy chain has the amino acid sequence shown in SEQ ID NO: 33, and the light chain has the amino acid sequence shown in SEQ ID NO: 41; 3) the heavy chain has the amino acid sequence shown in SEQ ID NO: 34, and the light chain has the amino acid sequence shown in SEQ ID NO: 42; 4) the heavy chain has the amino acid sequence shown in SEQ ID NO: 35, and the light chain has the amino acid sequence shown in SEQ ID NO: 42; 5) the heavy chain has the amino acid sequence shown in SEQ ID NO: 36, and the light chain has the amino acid sequence shown in SEQ ID NO: 42; 6) the heavy chain has the amino acid sequence shown in SEQ ID NO: 35, and the light chain has the amino acid sequence shown in SEQ ID NO: 43; 7) the heavy chain has the amino acid sequence shown in SEQ ID NO:36, and the light chain has the amino acid sequence shown in SEQ ID NO:43; 8) the heavy chain has the amino acid sequence shown in SEQ ID NO:38, and the light chain has the amino acid sequence shown in SEQ ID NO:41; 9) the heavy chain has the amino acid sequence shown in SEQ ID NO: 39, and the light chain has the amino acid sequence shown in SEQ ID NO: 44; 10) the heavy chain has the amino acid sequence shown in SEQ ID NO: 39, and the light chain has the amino acid sequence shown in SEQ ID NO: 45; 11) the heavy chain has the amino acid sequence shown in SEQ ID NO:39, and the light chain has the amino acid sequence shown in SEQ ID NO:46; 12) the heavy chain has the amino acid sequence shown in SEQ ID NO: 39, and the light chain has the amino acid sequence shown in SEQ ID NO: 47; 13) the heavy chain has the amino acid sequence shown in SEQ ID NO: 37, and the light chain has the amino acid sequence shown in SEQ ID NO: 43; 14) the heavy chain has the amino acid sequence shown in SEQ ID NO: 37, and the light chain has the amino acid sequence shown in SEQ ID NO: 44; 15) the heavy chain has the amino acid sequence shown in SEQ ID NO: 37, and the light chain has the amino acid sequence shown in SEQ ID NO: 47; 16) the heavy chain has the amino acid sequence shown in SEQ ID NO: 144, and the light chain has the amino acid sequence shown in SEQ ID NO: 159; 17) the heavy chain has the amino acid sequence shown in SEQ ID NO: 145, and the light chain has the amino acid sequence shown in SEQ ID NO: 160; 18) the heavy chain has the amino acid sequence shown in SEQ ID NO: 146, and the light chain has the amino acid sequence shown in SEQ ID NO: 161; 19) the heavy chain has the amino acid sequence shown in SEQ ID NO: 147, and the light chain has the amino acid sequence shown in SEQ ID NO: 162; 20) the heavy chain has the amino acid sequence shown in SEQ ID NO: 148, and the light chain has the amino acid sequence shown in SEQ ID NO: 163; 21) the heavy chain has the amino acid sequence shown in SEQ ID NO: 149, and the light chain has the amino acid sequence shown in SEQ ID NO: 164; 22) the heavy chain has the amino acid sequence shown in SEQ ID NO: 150, and the light chain has the amino acid sequence shown in SEQ ID NO: 165; 23) the heavy chain has the amino acid sequence shown in SEQ ID NO: 152, and the light chain has the amino acid sequence shown in SEQ ID NO: 167; 24) the heavy chain has the amino acid sequence shown in SEQ ID NO: 153, and the light chain has the amino acid sequence shown in SEQ ID NO: 168; 25) the heavy chain has the amino acid sequence shown in SEQ ID NO: 154, and the light chain has the amino acid sequence shown in SEQ ID NO: 169; 26) the heavy chain has the amino acid sequence shown in SEQ ID NO: 155, and the light chain has the amino acid sequence shown in SEQ ID NO: 170; 27) the heavy chain has the amino acid sequence shown in SEQ ID NO: 156, and the light chain has the amino acid sequence shown in SEQ ID NO: 171; 28) the heavy chain has the amino acid sequence shown in SEQ ID NO: 157, and the light chain has the amino acid sequence shown in SEQ ID NO: 172; or 29) The heavy chain has the amino acid sequence shown in SEQ ID NO:158, and the light chain has the amino acid sequence shown in SEQ ID NO:

173.

28. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-27, which is defucosylated.

29. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-28, which induces antibody-dependent cellular cytotoxicity (ADCC) in a subject upon administration.

30. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 29, which is capable of inducing activation of NK cells.

31. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 30, which is capable of inducing NK cell-mediated killing of tumor-infiltrating Treg cells in the subject.

32. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-31, which induces CCR8 internalization by tumor-infiltrating Treg cells.

33. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 32, which is a human antibody, a humanized antibody or a chimeric antibody.

34. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 33, which is an IgG1, IgG2, IgG3 or IgG4 isotype antibody or antigen binding fragment thereof.

35. The anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1-34, which is a monoclonal antibody, a bispecific antibody, a bispecific T cell engager (BiTE), a multispecific antibody, a biparatopic antibody, an immunoconjugate, an antibody drug conjugate, a chimeric antigen receptor (CAR), a T cell receptor (TCR) or any combination thereof.

36. The anti-CCR8 antibody or antigen binding portion thereof according to any one of claims 1 to 35, which targets the extracellular domain of human CCR8.

37. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-36, which binds to cells expressing CCR8 on their surface.

38. An anti-CCR8 antibody, or antigen binding portion thereof, according to any one of claims 1 to 37, which specifically binds to one or more amino acids within the N-terminal extracellular domain of CCR8.

39. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-38, wherein the cell is a regulatory T cell (Treg).

40. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-39, wherein the cell is a tumor infiltrating Treg.

41. The anti-CCR8 antibody, or antigen binding portion thereof, of any one of claims 1-40, which competes for binding with CCL1.

42. A pharmaceutical composition comprising the anti-CCR8 antibody or antigen-binding portion thereof according to any one of claims 1-41 and a pharmaceutically acceptable carrier.

43. A method of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject an anti-CCR8 antibody or antigen binding portion according to any one of claims 1-41 or a pharmaceutical composition according to claim 42.

44. The method of claim 43, wherein the disease or condition comprises a tumor.

45. The method of claim 44, wherein the tumor comprises Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic promyelocytic myelomonocytic monocytic erythroleukemia, chronic leukemia, chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt's lymphoma and marginal zone B-cell lymphoma, polycythemia vera lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's macroglobulinemia, Heavy chain disease, solid tumors, sarcomas and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon sarcoma, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma (H CC), liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular tumors, lung cancer, small cell lung cancer, non-small cell lung cancer, bladder tumors, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal cancer, esophageal cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) NS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, stomach cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell lung cancer, large cell lung cancer), melanoma, neuroblastoma; oral cancer (e.g., lip cancer, tongue cancer, mouth cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; respiratory system cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and urinary system cancer, and any combination thereof.

46. ​​The method of any one of claims 43 to 45, wherein the tumor is refractory or recurrent.

47. The method of any one of claims 43 to 46, wherein the tumor is advanced, locally advanced, or metastatic.

48. The method of any one of claims 43 to 47, further comprising administering an additional anti-cancer agent and / or method.

49. Use of an anti-CCR8 antibody, or antigen binding portion thereof, according to any one of claims 1 to 41 in the manufacture of a medicament for treating a disease or disorder.

50. The use according to claim 49, wherein the disease or disorder comprises a tumor.

51. The method of any one of claims 49-50, wherein the tumor comprises Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic promyelocytic myelomonocytic monocytic erythroleukemia, chronic leukemia, chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt's lymphoma and marginal zone B-cell lymphoma, polycythemia vera lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's giant glomerulus Proteinemia, Heavy chain disease, Solid tumors, Sarcomas and carcinomas, Fibrosarcoma, Myxosarcoma, Liposarcoma, Chondrosarcoma, Osteogenic sarcoma, Osteosarcoma, Chordoma, Angiosarcoma, Endotheliosarcoma, Lymphangiosarcoma, Lymphangioendotheliosarcoma, Synovialoma, Mesothelioma, Ewing's tumor, Leiomyosarcoma, Rhabdomyosarcoma, Sarcoma of the colon, Colorectal cancer, Pancreatic cancer, Breast cancer, Ovarian cancer, Prostate cancer, Squamous cell carcinoma, Basal cell carcinoma, Adenocarcinoma, Sweat gland carcinoma, Sebaceous gland carcinoma, Papillary carcinoma, Papillary adenocarcinoma, Cystadenocarcinoma, Medullary carcinoma, Bronchogenic carcinoma, Renal cell carcinoma, Hepatocellular carcinoma Carcinoma (HCC), liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular tumors, lung cancer, small cell lung cancer, non-small cell lung cancer, bladder tumors, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal carcinoma, esophageal cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system ( CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, stomach cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell lung cancer, large cell lung cancer), melanoma, neuroblastoma; oral cancer (e.g., lip cancer, tongue cancer, mouth cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; respiratory system cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and urinary system cancer, and any combination thereof.

52. The use according to any one of claims 49 to 51, wherein the tumor is refractory or recurrent.

53. The use according to any one of claims 49 to 51, wherein the tumor is advanced, locally advanced or metastatic.

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