Anti-CCR8 antibodies and uses thereof

Abstract

Provided herein are isolated monoclonal anti-CCR8 antibodies, their antigen-binding fragments, mutants, and variants, and their uses for treating diseases such as cancer. The mutant antibodies have altered complementarity-determining region 3 to remove post-translational modifications and improve antibody stability and efficacy. The variant antibodies have varying levels of defucosylation. Also provided herein are methods of using anti-CCR8 antibodies, including methods for treating cancer. Also provided are polynucleotides encoding the heavy or light chains or antigen-binding portions thereof described herein, and vectors, particularly expression vectors, containing the polynucleotides described herein.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of priority under Title 35, United States Code, Section 119(e) of U.S. Provisional Application No. 63 / 399,483, filed August 19, 2022. The disclosures of these prior applications are considered part of the disclosure of this application and are hereby incorporated by reference in their entireties into the disclosure of this application.

[0002] [Incorporating sequence table] The contents of the attached Sequence Listing are incorporated herein by reference. The attached Sequence Listing xml file, named 148640-004502_SL.xml, was created on August 18, 2023 and is 39,844 bytes.

[0003] [Summary of the Invention] The present invention relates to isolated anti-chemokine (C-C motif) receptor 8 (CCR8) monoclonal antibodies or antigen-binding fragments thereof, nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. Methods of producing the antibodies and methods of using the antibodies to treat diseases, including cancer and / or related complications, are also provided. [Background technology]

[0004] CCR8 is a chemokine receptor that mediates cell migration down a CCL1 or CCL18 gradient (Islam et al., JEM 210(10):1889-1898 (2013)). Recently, CCR8 has been identified as a highly specific cell surface marker for tumor-infiltrating regulatory T cells (TCRs) in human cancers. CCR8 expression is significantly higher on tumor-resident Tregs compared with circulating Tregs, whereas it is absent or very low on other T cell populations (cytotoxic T cells or effector T cells, respectively). Furthermore, CCR8 is predominantly expressed on highly immunosuppressive Tregs, which express FoxP3high, CD25high, TIGIT+, and LAG3+ and release high levels of IL-10 and TGF-β. Depletion of CCR8+ Tregs reduces immunosuppressive cytokines, modulates the tumor-promoting microenvironment, and restores antitumor immunity. Interestingly, in patients with breast or pancreatic cancer, high CCR8+ Treg numbers correlated with more advanced disease stage and decreased overall survival. Therefore, CCR8 is an ideal target for cancer immunotherapy to potentially treat and cure CCR8-positive cancers. Summary of the Invention

[0005] Aspects of the invention described herein relate to humanized antibodies directed against CCR8, mutants and variants thereof, and their use for the treatment of diseases such as cancer.

[0006] In one embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: HCVR including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15), wherein the antibody of the antigen-binding fragment specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided. In a specific embodiment herein, the antibody comprises an HCVR of SEQ ID NO: 36 and an LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody is Ab001_M4, and comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 2.

[0007] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: HC comprising a sequence having at least 80% identity to SEQ ID NO:6 and having the antigen-binding specificity thereof; and LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having its antigen-binding specificity; An isolated monoclonal antibody or antigen-binding fragment thereof is provided.

[0008] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: (a) HCVR, which includes: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQNYYYAMDY (SEQ ID NO: 16); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (b) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENNYYYAMDY (SEQ ID NO: 17); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (c) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENQYYYAMDY (SEQ ID NO: 18); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (d) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and HCVR CDR3 sequence having the amino acid sequence TRGSDQNYYYAMDY (SEQ ID NO: 20); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (e) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDNQYYYAMDY (SEQ ID NO: 21); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (f) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDQQYYYAMDY (SEQ ID NO: 22); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); wherein the antibody of the antigen-binding fragment specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided. In specific embodiments herein, the antibody is (a) an HCVR of SEQ ID NO: 33 and an LCVR of SEQ ID NO: 32; (b) an HCVR of SEQ ID NO: 34 and an LCVR of SEQ ID NO: 32; (c) an HCVR of SEQ ID NO: 35 and an LCVR of SEQ ID NO: 32; (d) an HCVR of SEQ ID NO: 37 and an LCVR of SEQ ID NO: 32; (e) an HCVR of SEQ ID NO: 38 and an LCVR of SEQ ID NO: 32, or (f) an HCVR of SEQ ID NO: 39 and an LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody comprises: (a) Ab001_M1, comprising a heavy chain of SEQ ID NO: 3 and a light chain of SEQ ID NO: 2; (b) Ab001_M2, comprising a heavy chain of SEQ ID NO: 4 and a light chain of SEQ ID NO: 2; (c) Ab001_M3, comprising a heavy chain of SEQ ID NO: 5 and a light chain of SEQ ID NO: 2; (d) Ab001_M5, comprising a heavy chain of SEQ ID NO: 7 and a light chain of SEQ ID NO: 2; (e) Ab001_M6, comprising a heavy chain of SEQ ID NO: 8 and a light chain of SEQ ID NO: 2; or (f) Ab001_M7, comprising a heavy chain of SEQ ID NO:9 and a light chain of SEQ ID NO:2.

[0009] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: (a) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 3 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (b) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 4 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (c) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 5 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (d) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 7 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (e) an HC comprising a sequence having at least 80% identity to SEQ ID NO:8 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO:2 and having the antigen-binding specificity thereof; or (f) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 9 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; wherein the antibody or antigen-binding fragment thereof specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided.

[0010] In one aspect, the HCVR sequence is SEQ ID NO: 33 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 34 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 35 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 37 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 38 and / or the LCVR sequence is SEQ ID NO: 32, or The HCVR sequence is SEQ ID NO: 39 and / or the LCVR sequence is SEQ ID NO: 32. The HCVR sequence is SEQ ID NO: 33, and the LCVR sequence is SEQ ID NO: 32. The HCVR sequence is SEQ ID NO: 34, and the LCVR sequence is SEQ ID NO: 32. The HCVR sequence is SEQ ID NO: 35, and the LCVR sequence is SEQ ID NO: 32. The HCVR sequence is SEQ ID NO: 37, and the LCVR sequence is SEQ ID NO: 32. the HCVR sequence is SEQ ID NO: 38 and the LCVR sequence is SEQ ID NO: 32, or The HCVR sequence is SEQ ID NO:39, and the LCVR sequence is SEQ ID NO:32.

[0011] In another aspect, the present invention provides an isolated monoclonal antibody comprising: (a) a heavy chain comprising SEQ ID NO:3 and a light chain comprising SEQ ID NO:2; (b) a heavy chain comprising SEQ ID NO:4 and a light chain comprising SEQ ID NO:2; (c) a heavy chain comprising SEQ ID NO:5 and a light chain comprising SEQ ID NO:2; (d) a heavy chain comprising SEQ ID NO:7 and a light chain comprising SEQ ID NO:2; (e) a heavy chain comprising SEQ ID NO:8 and a light chain comprising SEQ ID NO:2; (f) a heavy chain comprising SEQ ID NO:9 and a light chain comprising SEQ ID NO:2; wherein the antibody specifically binds to CCR8. Isolated monoclonal antibodies are provided.

[0012] In another embodiment, the antibody is a humanized antibody. In one embodiment, the antigen-binding fragment thereof is selected from the group consisting of Fab, Fab', F(ab'), F d , single-chain Fv or scFv, disulfide-linked F v, V-NAR domain, IgNar, intrabody, IgGACH2, minibody, F(ab')3, tetrabody, triabody, diabody, single domain antibody, DVD-Ig, Fcab, mAb2, (scFv)2, or scFv-Fc. In other embodiments, the monoclonal antibody or antigen-binding fragment thereof is K d binds to human CCR8 with an affinity of less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, 1 nM, 0.5 nM, or 0.01 nM. In one embodiment, the antibody induces antibody-dependent cell-mediated cytotoxicity (ADCC). In another embodiment, the antibody has a defucosylation level of at least 50%. In various embodiments, the antibody has a defucosylation level of at least 85%.

[0013] In another embodiment, the present invention provides a method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of any one of the isolated monoclonal antibodies or antigen-binding fragments described herein, thereby treating cancer in the subject.

[0014] In another embodiment, the present invention provides a method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of at least one isolated monoclonal antibody or antigen-binding fragment thereof described herein. In one embodiment, the method includes administering an antibody having: HCVR including: a CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); In a specific embodiment herein, the antibody comprises an HCVR of SEQ ID NO: 36 and an LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody is Ab001_M4, and comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 2.

[0015] In one embodiment, the cancer is a blood cancer. In some embodiments, the blood cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sézary syndrome, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma.

[0016] In another embodiment, the cancer is a solid tumor. In some embodiments, the solid tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colorectal cancer, renal cancer, urothelial cancer, ovarian cancer, endometrial cancer, uterine cancer, liver cancer, pancreatic cancer, bile duct cancer, gastric cancer, gastroesophageal cancer, esophageal cancer, cervical cancer, squamous cell carcinoma, prostate cancer, or bladder cancer. In one embodiment, the method further comprises administering to the patient a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition.

[0017] In additional embodiments, the invention provides polynucleotides encoding the heavy or light chain, or antigen-binding portion thereof, of any one of the antibodies or antigen-binding fragments thereof described herein.

[0018] In one embodiment, the present invention provides a vector comprising a polynucleotide described herein, The vector is an expression vector selected from the group consisting of a mammalian expression vector, a yeast expression vector, an insect expression vector, and a bacterial expression vector. A vector is provided. [Brief explanation of the drawings]

[0019] [Figure 1] 1 is a graph showing the ADCC potency of antibodies with various levels of defucosylation. [Figures 2A-2C] Figure 2 shows deamination of Ab001_Wt antibody. Figure 2A shows the results of a binding assay with CHO-hCCR8 cells. Figure 2B shows the results of an ADCC reporter assay. Figure 2C shows the results of peptide mapping. [Figure 3A-3B] Figure 3 shows the activity of Ab001_Wt antibody over time. Figure 3A shows the results of a binding assay over time. Figure 3B shows the results of an ADCC reporter assay over time. [Figures 4A-4C] Figure 4A shows the binding assay results for Ab001_M1-M4 on CHO.hCCR8 cells compared to Ab001_Wt. Figure 4B shows the binding assay results for Ab001_M3 and Ab001_M4 compared to Ab001_Wt. Figure 4C is a table showing the EC50s calculated in Figures 4A and 4B. [Figures 5A-5C]Figure 5A shows the ADCC reporter assay results for Ab001_M1 to Ab001_M4 compared with Ab001_Wt using CHO.hCCR8 as the target cells. Figure 5A shows the ADCC reporter assay results for Ab001_M1 and Ab001_M2 compared with Ab001_Wt. Figure 5B shows the ADCC reporter assay results for Ab001_M3 and Ab001_M4 compared with Ab001_Wt. Figure 5C is a table showing the EC50s calculated in Figures 5A and 5B. DETAILED DESCRIPTION OF THE INVENTION

[0020] Aspects of the invention described herein relate to humanized antibodies directed against CCR8 and their use for the treatment of diseases such as cancer.

[0021] Before the present compositions and methods are described, it is to be understood that this invention is not limited to the particular compositions, methods, and experimental conditions described, as such compositions, methods, and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0022] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise. Thus, for example, reference to a "method" includes one or more methods, and / or steps, of the type described herein that will become apparent to those skilled in the art upon reading this disclosure and so forth.

[0023] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, but it will be understood that modifications and variations are within the spirit and scope of this disclosure. Preferred methods and materials are described below.

[0025] Unless otherwise specified, all numerical values, such as concentrations or concentration ranges, described herein should be understood in all instances to be modified by the term "about." Thus, numerical values ​​typically include ±10% of the stated value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, the use of numerical ranges explicitly includes all possible subranges, and all individual numerical values ​​within that range (including integers and fractional portions of such values ​​within such ranges), unless the context clearly indicates otherwise.

[0026] Unless otherwise indicated, the term "at least" preceding a series of elements is understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0027] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," or "containing," or other variations thereof, are understood to mean the inclusion of the stated element or group of elements, but not the exclusion of other elements or groups of elements, and are intended to be non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or apparatus that includes a list of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent in such composition, mixture, process, method, article, or apparatus. Further, unless expressly stated to the contrary, "or" refers to an inclusive "or," not an exclusive "or." For example, condition A or B is satisfied if any of the following is true: A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), or both A and B are true (or exist).

[0028] As used herein, the conjunction "and / or" between multiple listed elements is understood to encompass both individual and multiple alternatives. For example, when two elements are connected by "and / or," the first alternative indicates that the first element may be applied without the second element. The second alternative indicates that the second element may be applied without the first element. The third alternative indicates that both the first and second elements may be applied. Any of these alternatives is understood to be within the meaning and therefore meet the requirements of the term "and / or" as used herein. It is also understood that multiple alternatives may be applied simultaneously and therefore meet the requirements of the term "and / or."

[0029] As used herein, the term "consists of," or variations such as "consist of" or "consisting of," as used throughout this specification and the claims, indicates that any listed element or group of elements is inclusive, but that no additional element or group of elements may be added to the specified method, structure, or composition.

[0030] As used herein, the term "consists essentially of," or variations such as "consist essentially of" or "consisting essentially of," as used throughout the specification and claims, indicates the inclusion of any recited element or group of elements, and the possible inclusion of any recited element or group of elements that do not materially alter the basic or novel characteristics of the specified method, structure, or composition. See MPEP § 2111.03.

[0031] The present invention recognizes the inherent disadvantages of the anti-CCR8 antibody disclosed in International Publication WO 2023 / 020621, specifically the antibody designated H11-10, which has a heavy chain of SEQ ID NO: 1 and a light chain of SEQ ID NO: 2. These disadvantages include the fact that asparaginyl residues in the antigen-binding complementarity-determining region (CDR) 3 within the HCVR are susceptible to deamination over time, resulting in reduced activity, and that fucosylation in the Fc region inhibits antibody-dependent cellular cytotoxicity (ADCC). Due to the importance of CDRs for antigen binding, any modification of these asparaginyl residues within the HCVR CDR3 may reduce the effectiveness of the antibody in engaging the CCR8 target.

[0032] In one embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: HCVR including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15), wherein the antibody of the antigen-binding fragment specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided. In a specific embodiment herein, the antibody comprises the HCVR of SEQ ID NO: 36 and / or the LCVR of SEQ ID NO: 32, particularly the HCVR of SEQ ID NO: 36 and the LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody is Ab001_M4, and comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 2.

[0033] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: HC comprising a sequence having at least 80% identity to SEQ ID NO:6 and having the antigen-binding specificity thereof; and LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having its antigen-binding specificity; An isolated monoclonal antibody or antigen-binding fragment thereof is provided.

[0034] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: (a) HCVR, which includes: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQNYYYAMDY (SEQ ID NO: 16); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (b) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENNYYYAMDY (SEQ ID NO: 17); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (c) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENQYYYAMDY (SEQ ID NO: 18); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (d) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and HCVR CDR3 sequence having the amino acid sequence TRGSDQNYYYAMDY (SEQ ID NO: 20); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (e) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDNQYYYAMDY (SEQ ID NO: 21); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (f) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDQQYYYAMDY (SEQ ID NO: 22); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); wherein the antibody of the antigen-binding fragment specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided. In specific embodiments herein, the antibody is (a) an HCVR of SEQ ID NO: 33 and an LCVR of SEQ ID NO: 32; (b) an HCVR of SEQ ID NO: 34 and an LCVR of SEQ ID NO: 32; (c) an HCVR of SEQ ID NO: 35 and an LCVR of SEQ ID NO: 32; (d) an HCVR of SEQ ID NO: 37 and an LCVR of SEQ ID NO: 32; (e) an HCVR of SEQ ID NO: 38 and an LCVR of SEQ ID NO: 32, or (f) an HCVR of SEQ ID NO: 39 and an LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody comprises: (a) Ab001_M1, comprising a heavy chain of SEQ ID NO: 3 and a light chain of SEQ ID NO: 2; (b) Ab001_M2, comprising a heavy chain of SEQ ID NO: 4 and a light chain of SEQ ID NO: 2; (c) Ab001_M3, comprising a heavy chain of SEQ ID NO: 5 and a light chain of SEQ ID NO: 2; (d) Ab001_M5, comprising a heavy chain of SEQ ID NO: 7 and a light chain of SEQ ID NO: 2; (e) Ab001_M6, comprising a heavy chain of SEQ ID NO: 8 and a light chain of SEQ ID NO: 2; or (f) Ab001_M7, comprising a heavy chain of SEQ ID NO:9 and a light chain of SEQ ID NO:2.

[0035] In another embodiment, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof comprising: (a) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 3 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (b) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 4 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (c) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 5 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (d) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 7 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (e) an HC comprising a sequence having at least 80% identity to SEQ ID NO:8 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO:2 and having the antigen-binding specificity thereof; or (f) an HC comprising a sequence having at least 80% identity to SEQ ID NO: 9 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; wherein the antibody or antigen-binding fragment thereof specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof is provided.

[0036] The terms "antibodies," "ab," "immunoglobulins," or "Ig," in the broadest sense, refer to glycoproteins with similar structural characteristics (i.e., molecules containing an antigen-binding site that immunospecifically binds to an antigen) and encompass a variety of antibody structures, including natural or artificial monovalent or polyvalent antibodies, including, but not limited to, monoclonal antibodies (including chimeric, humanized, and human monoclonal antibodies, particularly humanized monoclonal antibodies), polyclonal antibodies, single-chain antibodies, antibody fragments, and multispecific antibodies (e.g., bispecific antibodies). While antibodies exhibit binding specificity to a specific antigen, immunoglobulins include both antibodies and other antibody-like molecules that lack antigen specificity. "Antibody" encompasses any polypeptide containing an antigen-binding site, regardless of its source, species of origin, method of production, and properties. The term "antibody" may also broadly refer to a molecule comprising CDR1, CDR2, and CDR3 of a heavy chain and CDR1, CDR2, and CDR3 of a light chain, which molecule is capable of binding to an antigen. In some embodiments, an antibody comprises an HCVR and an LCVR. In some embodiments, an antibody comprises at least one heavy chain (HC) comprising an HCVR and at least a portion of a heavy chain constant region, and at least one light chain (LC) comprising an LCVR region and at least a portion of a light chain constant region. In some embodiments, an antibody comprises two heavy chains (each heavy chain comprising an HCVR and at least a portion of a heavy chain constant region) and two light chains (each light chain comprising an LCVR and at least a portion of a light chain constant region).

[0037] In some embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises: HCVR sequence of SEQ ID NO: 33 and / or LCVR sequence of SEQ ID NO: 32, HCVR sequence of SEQ ID NO: 34 and / or LCVR sequence of SEQ ID NO: 32, HCVR sequence of SEQ ID NO: 35 and / or LCVR sequence of SEQ ID NO: 32, HCVR sequence of SEQ ID NO: 37 and / or LCVR sequence of SEQ ID NO: 32, the HCVR sequence of SEQ ID NO: 8 and / or the LCVR sequence of SEQ ID NO: 32, or The HCVR sequence of SEQ ID NO: 39 and / or the LCVR sequence of SEQ ID NO: 32.

[0038] In other embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises: HCVR sequence of SEQ ID NO: 33 and LCVR sequence of SEQ ID NO: 32; HCVR sequence of SEQ ID NO: 34 and LCVR sequence of SEQ ID NO: 32; HCVR sequence of SEQ ID NO: 35 and LCVR sequence of SEQ ID NO: 32; HCVR sequence of SEQ ID NO: 37 and LCVR sequence of SEQ ID NO: 32; an HCVR sequence of SEQ ID NO: 38 and an LCVR sequence of SEQ ID NO: 32, or It includes the HCVR sequence of SEQ ID NO: 39 and the LCVR sequence of SEQ ID NO: 32.

[0039] "Native antibodies" and "intact immunoglobulins," etc., are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light chains (L chains) and two identical heavy chains (H chains). Light chains from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domains. Depending on the amino acid sequence of the constant domain of the heavy chain, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which may be further subdivided into subclasses (isotypes). For example, IgG antibodies include, but are not limited to, IgG1 (containing a γ1 constant region), IgG2 (containing a γ2 constant region), IgG3 (containing a γ3 constant region), and IgG4 (containing a γ4 constant region) antibodies; IgA antibodies include, but are not limited to, IgA1 (containing an α1 constant region) and IgA2 (containing an α2 constant region) antibodies; and IgM antibodies include, but are not limited to, IgM1 (containing a μ1 constant region) and IgM2 (containing a μ2 constant region). The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known.

[0040] An intact antibody may have one or more "effector functions," which refer to the biological activities attributable to the Fc region of the antibody (a native-sequence Fc region, or an amino acid sequence variant Fc region, or otherwise modified Fc region). Examples of antibody effector functions include: C1q binding; complement-dependent cytotoxicity; Fc receptor binding; ADCC; phagocytosis; down-regulation of cell surface receptors (e.g., B cell receptor (BCR)); and cross-presentation of antigens by antigen-presenting cells or dendritic cells.

[0041] Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide bonds varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain contains at one end a variable domain (V H ) followed by multiple constant domains. Each light chain has a variable domain (V L ) at one end and a constant domain at the other end; the light-chain constant domain aligns with the first constant domain of the heavy chain, and the light-chain variable domain aligns with the variable domain of the heavy chain. Specific amino acid residues are believed to form the interface between the light- and heavy-chain variable domains. Each variable region contains three segments called complementarity-determining regions (CDRs) or hypervariable regions, with the more highly conserved portions of the variable domains called framework regions (FRs). The heavy- and light-chain variable domains each contain four FR regions, which primarily adopt a β-sheet configuration and are linked by three CDRs. These CDRs form loops that connect and, in some cases, form part of the β-sheet structure. The CDRs of each chain are held in close proximity by the FRs and, together with the CDRs of the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., NIH Publ. No. 91-3242, Vol. I, pages 647-669

[1991] ). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity.

[0042] As used herein, the term "heavy chain variable region (HCVR)" refers to a region comprising at least CDR1 (CDR-H1), framework 2 (HFR2), CDR2 (CDR-H2), FR3 (HFR3), and CDR3 (CDR-H3) of the heavy chain. In some embodiments, the HCVR also comprises at least a portion (e.g., the entirety) of FR1 (HFR1) located N-terminal to CDR-H1 and / or at least a portion (e.g., the entirety) of FR4 (HFR4) located C-terminal to CDR-H3. In various embodiments, any HCVR sequence disclosed herein further comprises the HFR1 sequence of SEQ ID NO:23, the HFR2 sequence of SEQ ID NO:24, the HFR3 sequence of SEQ ID NO:25, and / or the HFR4 sequence of SEQ ID NO:26.

[0043] As used herein, the term "heavy chain constant region" refers to a region comprising at least three heavy chain constant domains, i.e., CH1, CH2, and CH3. Non-limiting exemplary heavy chain constant regions include gamma, delta, and alpha. Non-limiting exemplary heavy chain constant regions also include epsilon and mu. Each heavy chain constant region corresponds to an antibody isotype. For example, an antibody comprising a gamma constant region is an IgG antibody, an antibody comprising a delta constant region is an IgD antibody, an antibody comprising an alpha constant region is an IgA antibody, an antibody comprising an epsilon constant region is an IgE antibody, and an antibody comprising a mu constant region is an IgM antibody.

[0044] As used herein, the term "light chain variable region (LCVR)" refers to the region comprising CDR1 (CDR-L1), framework (FR) 2 (LFR2), CDR2 (CDR-L2), FR3 (LFR3), and CDR3 (CDR-L3) of the light chain. In some embodiments, the LCVR also comprises at least a portion (e.g., the entirety) of FR1 (LFR1) and / or at least a portion (e.g., the entirety) of FR4 (LFR4). In various embodiments, any LCVR sequence disclosed herein further comprises the LFR1 sequence of SEQ ID NO:27, the LFR2 sequence of SEQ ID NO:28, the LFR3 sequence of SEQ ID NO:29, and / or the LFR4 sequence of SEQ ID NO:30.

[0045] As used herein, the term "light chain constant region" refers to the region containing the light chain constant domain, i.e., CL. Non-limiting exemplary light chain constant regions include λ and κ.

[0046] As used herein, the term "light chain" refers to a polypeptide comprising at least an LCVR, with or without a leader sequence. In some embodiments, a light chain comprises at least a portion of a light chain constant region. As used herein, the term "full-length light chain" refers to a polypeptide comprising an LCVR and a light chain constant region, with or without a leader sequence.

[0047] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies; i.e., the individual antibodies comprising the population are identical except for minor amounts of naturally occurring mutations. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, unlike conventional (polyclonal) antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The "monoclonal antibodies" may also be isolated from phage antibody libraries using the techniques described in, e.g., Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991).

[0048] In a general aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof that binds to chemokine (CC motif) receptor 8 (CCR8).

[0049] As used herein, an antibody that "specifically binds to CCR8" refers to an antibody and / or antigen-binding domain that binds to CCR8, preferably human CCR8, and has a KD of 1×10 -7 M or less, preferably 1 × 10-8 M or less, preferably 5×10 -9 M or less, 1×10 -9 M or less, 5×10 -10 M or less, or 1 x 10 -10 In one embodiment, the antibody and / or antigen-binding domain binds to cynomolgus CCR8. The term "KD" refers to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M). The KD value of an antibody can be determined using methods known in the art in light of the present disclosure. For example, the KD of an antibody can be determined by using surface plasmon resonance (e.g., by using a biosensor system, such as a Biacore® system) or by using biolayer interferometry technology (e.g., an Octet RED96 system).

[0050] The smaller the KD value of an antibody, the higher the affinity with which the antibody binds to the target antigen.

[0051] As used herein, "IC 50 The term "IC" refers to the half-maximal inhibitory concentration of a monoclonal or bispecific antibody, or antigen-binding fragment thereof, of the invention. 50 is a measure of the potency of a monoclonal or bispecific antibody, or antigen-binding fragment thereof, of the invention for inhibiting the binding of CCL1 to CCR8 or for inhibiting the function of CCR8 in cells. In one embodiment, the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, has a KD of about 10 -7 Less than M, about 10 -8 Less than M, about 10 -9 Less than M, about 10 -10 Less than M, about 10 -11 Less than M, about 10 -12 Less than M or about 10 -13 It is less than M.

[0052] As used herein, "EC 50The term "EC" refers to the half maximal effective concentration of a monoclonal or bispecific antibody, or antigen-binding fragment thereof, of the invention. 50 refers to the concentration of a monoclonal or bispecific antibody, or antigen-binding fragment thereof, to induce a biological response (e.g., cell death) that is exactly halfway between the baseline and maximum over a specific exposure time. In certain embodiments, the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, has an EC 50 is less than about 1 μM, about 1000 nM to about 100 nM, about 100 nM to about 10 nM, about 10 nM to about 1 nM, about 1000 pM to about 500 pM, about 500 pM to about 200 pM, less than about 200 pM, about 200 pM to about 150 pM, about 200 pM to about 100 pM, about 100 pM to about 10 pM, or about 10 pM to about 1 pM.

[0053] In one aspect, the HCVR sequence is SEQ ID NO: 33 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 34 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 35 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 36 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 37 and / or the LCVR sequence is SEQ ID NO: 32; the HCVR sequence is SEQ ID NO: 38 and / or the LCVR sequence is SEQ ID NO: 32, or The HCVR sequence is SEQ ID NO:39 and / or the LCVR sequence is SEQ ID NO:32.

[0054] In another aspect, the present invention provides an isolated monoclonal antibody comprising: (a) a heavy chain comprising SEQ ID NO:3 and a light chain comprising SEQ ID NO:2; (b) a heavy chain comprising SEQ ID NO:4 and a light chain comprising SEQ ID NO:2; (c) a heavy chain comprising SEQ ID NO:5 and a light chain comprising SEQ ID NO:2; (d) a heavy chain comprising SEQ ID NO:7 and a light chain comprising SEQ ID NO:2; (e) a heavy chain comprising SEQ ID NO:8 and a light chain comprising SEQ ID NO:2; (f) a heavy chain comprising SEQ ID NO:9 and a light chain comprising SEQ ID NO:2; wherein the antibody specifically binds to CCR8. Isolated monoclonal antibodies are provided.

[0055] In one embodiment, the antibody is a humanized antibody. "Humanized" forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from the non-human immunoglobulin. In most cases, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from the recipient CDRs are replaced by residues from the CDRs of a non-human species (donor antibody) such as mouse, rat, or rabbit that has the desired specificity, affinity, and binding capacity. In some cases, residues from the Fv FRs of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can contain residues that are found neither in the recipient antibody nor in the imported CDR or FR sequences. These modifications are made to further optimize and maximize antibody performance. Generally, a humanized antibody will comprise substantially the entirety of at least one, and usually two, variable domains, in which all or substantially all of the CDRs correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody also optimally comprises at least a portion of an immunoglobulin constant region (Fc), usually that of a human immunoglobulin. For details, see Jones et al., Nature, 321:522-525 (1986); Reichmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992). Humanized antibodies include PRIMATIZED™ antibodies, whose antigen-binding regions are derived from antibodies produced by immunizing macaque monkeys with an antigen of interest.

[0056] Methods for humanizing non-human antibodies are well known in the art. Preferably, a humanized antibody has one or more amino acid residues introduced into it from a non-human source. These non-human amino acid residues, often referred to as "import" residues, are usually taken from an "import" variable domain. Humanization can be performed essentially according to the method of Winter et al. (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536

[1988] ), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Such "humanized" antibodies are therefore chimeric antibodies (U.S. Pat. No. 4,816,567), in which substantially less of the corresponding sequence from the non-human species has been substituted compared to the intact human variable domain. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.

[0057] The selection of human variable domains (light and heavy chains) used in creating a humanized antibody is crucial to reducing antigenicity. According to the so-called "best-fit" method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable domain sequences. The human sequence closest to the rodent sequence is then used as the human FR for the humanized antibody (Sims et al., J. Immunol., 151:2296 (1993); Chothia et al., J. Mol. Biol., 196:901

[1987] ). Another method uses specific FRs derived from the consensus sequence of all human antibodies belonging to a particular light or heavy chain subgroup. The same FRs can be used for multiple different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623

[1993] ).

[0058] It is further important that antibodies be humanized with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, according to a preferred method, humanized antibodies are prepared by a process of analysis of the parental sequences and various theoretical humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display predicted three-dimensional conformations of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the function of the candidate immunoglobulin sequence, i.e., analysis of residues that influence the ability of the candidate immunoglobulin to bind to its antigen. In this way, FR residues from the recipient and import sequences can be selected and combined to achieve desired antibody characteristics, such as improved affinity for the target antigen(s). In general, CDR residues are directly and most significantly involved in influencing antigen binding.

[0059] Alternatively, it is now possible to generate transgenic animals (e.g., mice) that can be immunized to produce a full repertoire of human antibodies without producing endogenous immunoglobulins. For example, it has been reported that homozygous deletion of the antibody heavy-chain joining region (JH) gene in chimeric and germline mutant mice completely inhibits endogenous antibody production. Introduction of the human germline immunoglobulin gene array into such germline mutant mice results in the production of human antibodies upon antigen stimulation. See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al., Year in Immuno., 7:33 (1993). Human antibodies may also be derived from phage display libraries (Hoogenboom et al., J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581-597

[1991] ).

[0060] Antibodies can be humanized by replacing Fv variable region sequences not directly involved in antigen binding with equivalent sequences from human Fv variable regions. General reviews of humanized chimeric antibodies are described in Morrison et al. (Science 229:1202-1207 (1985)) and Oi et al. (BioTechniques 4:214 (1986)). These methods include isolating, manipulating, and expressing nucleic acid sequences encoding all or part of immunoglobulin Fv variable regions from at least one of the heavy or light chains. Sources of such nucleic acids are well known to those of skill in the art and can be obtained, for example, from antibody-producing hybridomas. Recombinant DNA encoding a humanized or chimeric antibody, or a fragment thereof, can then be cloned into an appropriate expression vector.

[0061] Alternatively, humanized antibodies can be produced by CDR substitution (U.S. Patent No. 5,225,539; Jones, Nature 321:552-525 (1986); Verhoeyan et al., Science 239:1534 (1988); and Beidler, J. Immunol. 141:4053-4060 (1988)).

[0062] In another embodiment, the antigen-binding fragment thereof is a Fab, Fab', F(ab'), F d , single-chain Fv or scFv, disulfide-linked F v , V-NAR domain, IgNar, intrabody, IgGACH2, minibody, F(ab')3, tetrabody, triabody, diabody, single domain antibody, DVD-Ig, Fcab, mAb2, (scFv)2, or scFv-Fc.

[0063] Experimentally, antibodies can be cleaved with the protease papain, which cleaves each heavy chain and generates three separate antibody fragments. "Antibody fragments" include portions of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', and F(ab')2, Fc fragments or Fc fusion products, single-chain Fv (scFv), disulfide-linked Fv (sdfv), and fragments containing the VL or VH domain; diabodies, tribodies, etc. (Zapata et al. Protein Eng. 8(10):1057-1062

[1995] ). The terms "antibody fragment" or "antigen-binding portion" (of an antibody) include, but are not limited to, fragments capable of binding to antigen. A two-unit unit consisting of a light chain and a fragment of the heavy chain approximately equal in mass to the light chain is called a Fab fragment (i.e., an "antigen-binding" fragment). The third unit consists of two equal segments of heavy chains and is called the Fc fragment. The Fc fragment is not normally involved in antigen-antibody binding, but is important in later processes involved in removing the antigen from the body.

[0064] The Fab fragment contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. "Fab" refers to an antibody fragment with a molecular weight of approximately 50,000 daltons and antigen-binding activity. It contains approximately the N-terminal half of the heavy chain and the entire light chain, linked by disulfide bridges. Fab can be specifically obtained by treating immunoglobulins with the protease papain. Fab' fragments differ from Fab fragments in that several residues, including one or more cysteines from the antibody hinge region, are added to the carboxy terminus of the heavy chain CH1 domain. Fab'-SH, as used herein, refers to Fab' in which the cysteine ​​residue(s) in the constant domains have free thiol groups.

[0065] F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical linkages of antibody fragments are also known. The term "F(ab')2" refers to a fragment of approximately 100,000 daltons that retains antigen-binding activity. This fragment is slightly larger than two Fab fragments linked via disulfide bridges in the hinge region. These fragments can be obtained by treating immunoglobulins with the protease pepsin. Fab fragments can be obtained from F(ab')2 fragments by cleaving the disulfide bridges in the hinge region.

[0066] The Fc region of an antibody is the tail region of the antibody, which interacts with cell surface receptors and certain proteins of the complement system. This property allows antibodies to activate the immune system. In antibody isotypes IgG, IgA, and IgD, the Fc region consists of two identical protein fragments derived from the second and third constant domains of the antibody's two heavy chains; in IgM and IgE, the Fc region contains three heavy chain constant domains (CH domains 2-4) on each polypeptide chain. The IgG Fc region contains highly conserved N-glycosylation sites. Glycosylation of the Fc fragment is essential for Fc receptor-mediated activity. N-glycans attached to this site are primarily core-fucosylated, complex-type, biantennary structures. In addition, a small amount of these N-glycans also bear bisecting GlcNAc and α-2,6-linked sialic acid residues.

[0067] Fc fusion proteins (also known as Fc chimeric fusion proteins, Fc-Ig, Ig-based chimeric fusion proteins, and Fc-tagged proteins) are genetically linked peptides or proteins of interest to the Fc domain of IgG. Fc fusion proteins have become valuable reagents for in vivo and in vitro research. The binding partners fused to the Fc domain can range from single peptides, ligands activated by binding to cell surface receptors, signaling molecules, extracellular domains of receptors activated by dimerization, or bait proteins used to identify binding partners in protein microarrays. One of the most important features of the Fc domain in vivo is its ability to significantly extend the plasma half-life of proteins of interest, which can improve the therapeutic efficacy of biotherapeutics; making Fc fusion proteins attractive biotherapeutics.

[0068] The Fc fusion protein can be part of a pharmaceutical composition comprising the Fc fusion protein and a pharmaceutically acceptable carrier excipient or carrier. Pharmaceutically acceptable carriers, excipients, or stabilizers are well known in the art (Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980)). Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed and can include: buffers (e.g., phosphate, citric acid, and other organic acids); antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins (e.g., , serum albumin, gelatin, or immunoglobulin); hydrophilic polymers (e.g., polyvinylpyrrolidone); amino acids (e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine); monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrin); chelating agents (e.g., EDTA); sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions (e.g., sodium); metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants (e.g., TWEEN™, PLURONICS™, or polyethylene glycol (PEG)).

[0069] An "Fv" is the minimum antibody fragment containing a complete antigen-recognition and binding site. An "scFv" dimer corresponds to two scFv molecules linked by a peptide bond. This Fv chain is often the result of expression of a fusion gene in which the VH and VL encoding genes are connected by a peptide-encoding linker sequence. Human scFv fragments may contain CDR regions that maintain the proper three-dimensional structure, preferably by means of recombinant DNA technology. This region consists of a dimer of one heavy-chain variable domain and one light-chain variable domain in tight, non-covalent association. In this structure, the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific CDRs) has the ability to recognize and bind antigen, although with lower affinity than the entire binding site.

[0070] "Single-chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Preferably, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the sFv to form the desired structure for antigen binding. For a review of sFvs, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0071] A "dsFv" fragment is a VH-VL heterodimer stabilized by disulfide bridges; it can be bivalent (dsFv2). Divalent sc(Fv)2 or multivalent antibody fragments can be formed by spontaneous association of monovalent scFvs or can be produced by linking scFv fragments via peptide bond sequences.

[0072] The Fc fragment is responsible for the biological properties of the antibody, specifically the ability to be recognized by immune effectors or to activate complement. It consists of the constant fragment of the heavy chain beyond the hinge region.

[0073] "Diabodies" refer to small antibody fragments with two antigen-fixing sites. These fragments contain a variable heavy domain, VH, linked to a variable light domain, VL, in the same VH-VL polypeptide chain. The two domains use a linker sequence that is too short to pair within the same chain, thereby forcing pairing with two complementary domains on separate chains, thereby generating two antigen-fixing sites.

[0074] Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were obtained by proteolysis of intact antibodies (see, e.g., Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992) and Brennan et al., Science, 229:81

[1985] ). However, these fragments can now be produced directly by recombinant host cells. For example, antibody fragments can be isolated from the antibody phage libraries discussed above. Alternatively, Fab′-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab′)2 fragments (Carter et al., Bio / Technology 10:163-167

[1992] ). Alternatively, F(ab′)2 fragments can be directly isolated from recombinant host cell culture. Other techniques for producing antibody fragments will be apparent to those skilled in the art. In other embodiments, the antibody of choice is a single chain Fv fragment (scFv). See WO93 / 16185.

[0075] In other embodiments, the monoclonal antibody or antigen-binding fragment thereof is dbinds to human CCR8 with less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, 1 nM, 0.5 nM, or 0.01 nM.

[0076] The term "antigen-binding domain" refers to the portion of an antibody molecule that comprises the area that specifically binds to or is complementary to part or all of an antigen. If the antigen is large, an antibody may bind only to a specific portion of the antigen. An "epitope" or "antigenic determinant" is the portion of an antigen molecule that is responsible for interacting with the antigen-binding domain of an antibody. An antigen-binding domain may be provided by one or more antibody variable domains (e.g., a so-called Fd antibody fragment consisting of a VH domain). An antigen-binding domain may comprise an antibody LCVR and an antibody HCVR.

[0077] According to the present invention, an "antigen" includes any substance that elicits an immune response. Specifically, "antigen" relates to any substance, preferably a peptide or protein, that specifically reacts with antibodies or T lymphocytes (T cells). According to the present invention, the term "antigen" includes any molecule that contains at least one epitope. Preferably, an antigen in the context of the present invention is a molecule that induces an immune response (after processing, if necessary). According to the present invention, any suitable antigen that is a candidate for an immune response may be used, where the immune response is preferably a cellular immune response. In the context of embodiments of the present invention, the antigen is preferably presented by cells, preferably antigen-presenting cells (including diseased cells, particularly cancer cells), in the context of MHC molecules, which elicit an immune response against the antigen. The antigen is preferably a product corresponding to or derived from a natural antigen. Such natural antigens include tumor antigens.

[0078] The term "epitope" refers to an antigenic determinant on a molecule such as an antigen, i.e., a portion or fragment of a molecule that is recognized by the immune system. An epitope of a protein, such as a tumor antigen, preferably comprises a continuous or discontinuous portion of said protein.

[0079] The terms "epitope", "antigen peptide", "antigen epitope", "immunogenic peptide", and "MHC binding peptide" may be used interchangeably herein and are preferably associated with an incomplete phenotype of an antigen and are preferably capable of eliciting an immune response against the antigen or against cells that express or contain, and preferably present, the antigen.

[0080] The term "binding affinity" generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner. Various methods for measuring binding affinity or avidity are known in the art, any of which may be used for purposes of the present invention. Specific exemplary embodiments are described below.

[0081] As used herein, "specific binding" refers to the binding of an antibody to a predetermined antigen. Antibodies typically bind to antibodies at a concentration of about 10 -8 K below M D and has an affinity (K) that is at least 10-fold lower, preferably at least 100-fold lower, for the predetermined antigen compared to the binding affinity for a nonspecific antigen other than the predetermined antigen or a closely related antigen (e.g., BSA, casein). D Alternatively, the antibody binds at about 10 6 M -1 , or about 10 7 M -1 , or about 10 8 M -1 , or 10 9 M -1 or higher K Aand may bind with an affinity corresponding to at least 10-fold greater, preferably at least 100-fold greater, affinity (K) for a predetermined antigen compared to the binding affinity for a nonspecific antigen other than the predetermined antigen or a closely related antigen (e.g., BSA, casein). A The bond is expressed by

[0082] "k d ”(seconds -1 The term "K", as used herein, is intended to refer to the dissociation rate constant of a particular antibody-antigen interaction. This value is also referred to as the "off value." D " (M -1 The term ) as used herein is intended to refer to the dissociation equilibrium constant of a particular antibody-antigen interaction.

[0083] In some embodiments, the CCR8-binding antibodies of the invention have an affinity for CCR8 (e.g., for human CCR8) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 Dissociation constant (K d In certain embodiments, the CCR8-binding antibody has a CCR8 binding affinity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 Dissociation constant (K d )

[0084] "k a " (M -1 seconds -1The term "K", as used herein, is intended to refer to the binding rate constant of a particular antibody-antigen interaction. A The term "" (M), as used herein, is intended to refer to the binding equilibrium constant of a particular antibody-antigen interaction.

[0085] As described herein, anti-CCR8 antibodies are also referred to as wild-type anti-CCR8 antibodies (having an HCVR having the sequence of SEQ ID NO: 31 and / or an LCVR having the sequence of SEQ ID NO: 32) and mutants thereof. As used herein, the term "mutant" antibody refers to an antibody having a mutation in CDR3 to remove post-translational modifications (deamination and isomerization) of the asparaginyl residues N104 and / or N105 in the wild-type sequence. Mutation of the three amino acid sequence (DNN) at positions 103-105 reduces or prevents deamination and isomerization, thereby improving the stability and potency of the antibody. In particular, mutation of the DNN sequence to EQQ prevents deamination and maintains CCR8 binding equivalent to that of the parent antibody. This modification significantly extends the active half-life of the mutant antibody compared to that of the parent antibody.

[0086] An antibody having an HCVR sequence of any one of SEQ ID NOs: 33 to 39 and an LCVR sequence of SEQ ID NO: 32 is referred to as a mutant anti-CCR8 antibody. In particular, an antibody having an HCVR sequence of SEQ ID NO: 36 and an LCVR sequence of SEQ ID NO: 32 is a preferred mutant anti-CCR8 antibody.

[0087] In one embodiment, the antibody induces antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC is a mechanism of cell-mediated immune defense in which effector cells of the immune system actively lyse target cells whose surface antigens are bound by specific antibodies. It is one mechanism by which antibodies can act to limit and contain infection as part of the humoral immune response. ADCC is independent of the immune complement system, which also lyses targets but does not require other cells. ADCC requires effector cells, classically known as natural killer (NK) cells, which typically interact with immunoglobulin G (IgG) antibodies. However, macrophages, neutrophils, and eosinophils can also mediate ADCC; for example, eosinophils kill certain parasites known as helminths via IgE antibodies. In general, ADCC is typically described as an immune response against antibody-attached cells, ultimately leading to the lysis of infected or non-host cells. In recent literature, the therapeutic importance of cancer cells and deeper insights into their deceptively complex pathways have attracted increasing interest among medical researchers.

[0088] As used herein, the term "variant" antibody refers to an antibody with a different level of fucosylation. A defucosylated monoclonal antibody is a monoclonal antibody produced in modified cells or under specific conditions, in which the oligosaccharides in the Fc region of the antibody have reduced or absent fucose units compared to antibodies produced in unmodified cells or under normal conditions. Defucosylated antibodies increase ADCC. Most approved monoclonal antibodies belong to the IgG1 isotype, which has two N-linked biantennary complex oligosaccharides attached to its Fc region. The Fc region exerts ADCC effector function through interaction with leukocyte receptors belonging to the FcγR family. ADCC is important for the efficacy of cancer antibodies, but many approved cancer antibodies exhibit lower than desired ADCC due to nonspecific IgG competing with the drug for binding to FcγRIIIa on natural killer cells. Defucosylated monoclonal antibodies overcome this issue through improved FcγRIIIa binding.

[0089] In a review by Pereira et al. (MABS 2018, VOL. 10, NO. 5, 693-711), the relationship between antibody core fucosylation and ADCC, various strategies for producing defucosylated antibodies, and the latest information on defucosylated antibody drugs currently in clinical trials and approved are discussed. The disclosure of this article is incorporated herein in its entirety.

[0090] In another embodiment, the antibody has a defucosylation level of at least 50%, e.g., the antibody has a defucosylation level of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more.

[0091] In various embodiments, the antibody has a defucosylation level of at least 85%, e.g., the antibody has a defucosylation level of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.

[0092] In another embodiment, the present invention provides a method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of any one of the isolated monoclonal antibodies or antigen-binding fragments described herein, thereby treating cancer in the subject.

[0093] In another embodiment, the present invention provides a method of treating cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of at least one isolated monoclonal antibody or antigen-binding fragment thereof described herein, thereby treating cancer in the subject. In one embodiment, the method comprises administering an antibody having: HCVR including: a CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); In a specific embodiment herein, the antibody comprises an HCVR of SEQ ID NO: 36 and an LCVR of SEQ ID NO: 32. In a specific embodiment herein, the antibody is Ab001_M4, and comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 2.

[0094] The term "subject" as used herein refers to any individual or patient that the subject method is carried out.Generally, the subject is human, but as those skilled in the art will understand, the subject can also be an animal.Therefore, the definition of subject includes vertebrates, such as rodents (including mice, rats, hamsters and guinea pigs), cats, dogs, rabbits, livestock (including cows, horses, goats, sheep, pigs, chickens, etc.), and other animals, including primates (including monkeys, chimpanzees, orangutans and gorillas).

[0095] The term "treatment" is used interchangeably herein with the term "therapeutic method" and refers to both: 1) a therapeutic procedure or measure that cures, slows progression, alleviates symptoms, and / or halts the progression of a diagnosed pathological condition or disorder, and 2) a prophylactic / preventative measure. Those in need of treatment can include individuals who already have a particular medical condition, as well as those who may eventually develop the disorder (i.e., those requiring preventative measures). "Treatment" encompasses any administration or application of a therapeutic agent to a disease (also referred to herein as a "disorder" or "condition") in a mammal, including a human, and also includes inhibiting a disease or the progression of a disease, inhibiting or slowing a disease or its progression, halting its progression, partially or completely alleviating a disease, partially or completely alleviating one or more symptoms of a disease, or restoring or repairing a lost, deficient, or defective function; or stimulating an inefficient process. The term "treatment" also includes reducing the severity of any phenotypic trait and / or reducing the incidence, degree, or likelihood of that trait.

[0096] The terms "therapeutically effective amount," "effective dose," "therapeutically effective dose," "effective amount," and the like refer to an amount of the compound of interest that elicits the biological or medical response in a tissue, system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician. Generally, the response is either an improvement in the patient's symptoms or a desired biological result (e.g., treatment of cancer). The therapeutically effective amount of the CCR8 antagonist of the present invention may vary depending on factors such as the disease state, the individual's age, sex, and weight, and the ability of the antagonist to elicit a desired response in an individual. A therapeutically effective amount encompasses an amount in which any toxic or detrimental effects of the CCR8 antagonist are outweighed by the therapeutically beneficial effects.

[0097] The terms "administration of" and / or "administering" should be understood to mean providing a therapeutically effective amount of a pharmaceutical composition to a subject in need of treatment. The route of administration can be enteral, topical, or parenteral. Thus, routes of administration include, but are not limited to, intradermal, subcutaneous, intravenous, intraperitoneal, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal (intradermal), transdermal, intratracheal, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal, oral, sublingual buccal, rectal, vaginal, nasal ocular administration, and infusion, inhalation, and nebulization. As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration. In various embodiments, the anti-CCR8 antibody is administered subcutaneously or intravenously.

[0098] The antibodies and antigen-binding fragments thereof described herein can be prepared as pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof and a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid, or carrier commonly used in the art, which is used with a therapeutic agent to form a "pharmaceutical composition" and be administered to a subject. A pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations used and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is appropriate for the formulation being used. For example, if the therapeutic agent is administered orally, the carrier may be a gel capsule. If the therapeutic agent is administered subcutaneously, the carrier ideally does not irritate the skin or cause injection site reactions.

[0099] Pharmaceutical compositions may be administered in a variety of unit dosage forms depending on the method of administration. Suitable unit dosage forms include, but are not limited to, powders, tablets, pills, capsules, troches, suppositories, patches, nasal sprays, injections, implantable sustained-release formulations, lipid complexes, and the like.

[0100] The subject compositions may be formulated as solid, semi-solid, liquid, or gaseous form preparations, including, but not limited to, tablets, capsules, powders, granules, ointments, solutions, suppositories, enemas, injections, inhalants, and aerosols.

[0101] In various embodiments, compositions containing CCR8 antagonists are provided as formulations using a wide variety of pharmaceutically acceptable carriers (see, for example, Gennaro, Drugfacts Plus, 20th ed. (2003); Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ed., Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook of Pharmaceutical Excipients, 3rd ed., Pharmaceutical Press (2000)). A variety of pharmaceutically acceptable carriers, including vehicles, adjuvants, and diluents, can be used. In addition, a variety of pharmaceutically acceptable auxiliary substances, such as pH adjusting agents and buffers, tonicity adjusting agents, stabilizers, wetting agents, etc., can also be used. Non-limiting examples of carriers include saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof.

[0102] In various embodiments, compositions comprising a CCR8 antagonist can be formulated for injection, including subcutaneous administration, by dissolving, suspending, or emulsifying in an aqueous or non-aqueous solvent (e.g., vegetable oil or other oil, synthetic fatty acid glyceride, higher fatty acid ester, or propylene glycol); and, if necessary, with conventional additives (e.g., solubilizers, isotonicity adjusting agents, suspending agents, emulsifiers, stabilizers, and preservatives).

[0103] In various embodiments, the compositions can be formulated for inhalation using, for example, pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0104] In various embodiments, the compositions can also be formulated as sustained-release microcapsules, for example, using biodegradable or non-biodegradable polymers. Non-limiting examples of biodegradable formulations include polylactic-co-glycolic acid (PLGA) polymers. Non-limiting examples of non-biodegradable formulations include polyglycerol fatty acid esters. Some methods for making such formulations are described, for example, in EP 1125584 A1.

[0105] Pharmaceutical dosage packs are also provided, each containing one or more doses of an anti-CCR8 antibody and its antigen-binding fragment. In some embodiments, a unit dose is provided, the unit dose containing a predetermined amount of a composition comprising an anti-CCR8 antibody and its antigen-binding fragment, with or without one or more additives. In some embodiments, such a unit dose is supplied in a single-use pre-filled syringe for injection. In various embodiments, the composition contained in the unit dose may include saline, sucrose, or the like; a buffer such as phosphate; and / or may be formulated within a stable and effective pH range. Alternatively, in some embodiments, the composition may be provided as a lyophilized powder that can be reconstituted by adding an appropriate liquid (e.g., sterile water). In some embodiments, the composition includes one or more substances that inhibit protein aggregation, including, but not limited to, sucrose and arginine. In some embodiments, the compositions of the present invention include heparin and / or proteoglycan.

[0106] The pharmaceutical compositions are administered in an amount effective for the treatment or prevention of a particular indication. A therapeutically effective amount typically depends on the weight of the subject being treated, their physical or health condition, the extent of the condition being treated, or the age of the subject being treated.

[0107] In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 50 μg / kg body weight to about 50 mg / kg body weight per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 100 μg / kg body weight to about 50 mg / kg body weight per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 100 μg / kg body weight to about 20 mg / kg body weight per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 0.5 mg / kg body weight to about 20 mg / kg body weight per administration.

[0108] In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 10 mg to about 1000 mg per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 20 mg to about 500 mg per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 20 mg to about 300 mg per administration. In some embodiments, anti-CCR8 antibodies and antigen-binding fragments thereof may be administered in an amount ranging from about 20 mg to about 200 mg per administration.

[0109] The anti-CCR8 antibody and antigen-binding fragment thereof composition can be administered to a subject as needed. In some embodiments, an effective dose of the anti-CCR8 antibody and antigen-binding fragment thereof is administered to a subject one or more times. In various embodiments, an effective dose of the anti-CCR8 antibody and antigen-binding fragment thereof is administered to a subject monthly, or less than monthly (e.g., every two months, every three months, or every six months, etc.). In other embodiments, an effective dose of the anti-CCR8 antibody and antigen-binding fragment thereof is administered more than monthly (e.g., every two weeks, every week, twice a week, three times a week, once a day, or multiple times a day, etc.). An effective dose of the anti-CCR8 antibody and antigen-binding fragment thereof is administered to a subject at least once. In some embodiments, an effective dose of the CCR8 antagonist can be administered multiple times, such as over a period of at least one month, at least six months, or at least one year. In some embodiments, the anti-CCR8 antibody and antigen-binding fragment thereof is administered to a subject as needed to alleviate one or more symptoms of a condition.

[0110] The antibodies described herein can be used for the treatment of cancer.

[0111] Cancer is a group of diseases that involve abnormal cell growth and can invade or metastasize to other parts of the body. In 2015, approximately 90.5 million people suffered from cancer, with approximately 14.1 million new cases occurring annually and approximately 8.8 million deaths (15.7% of deaths). The most common types of cancer in men are lung, prostate, colon, and stomach cancer. The most common types in women are breast, colon, lung, and cervical cancer.

[0112] The term "cancer" refers to a group of diseases characterized by abnormal and uncontrolled cell growth, which distinguishes cancers (malignant tumors) from benign tumors by their potential to initiate at one site (the primary site) and invade and metastasize to other sites (metastatic sites, or metastases). Virtually every organ can be affected, resulting in over 100 different types of cancer that can affect humans. Cancer can be caused by many factors, including: genetic predisposition, viral infection, exposure to ionizing radiation, exposure to environmental pollutants, smoking and / or alcohol use, obesity, poor diet, lack of exercise, or any combination thereof.

[0113] As used herein, "neoplasm" or "tumor" (including grammatical variations thereof) refers to a new, abnormal growth of tissue, which may be benign or cancerous. In related embodiments, neoplasm refers to a neoplastic disease or disorder, including, but not limited to, various cancers. For example, such cancers may include: prostate cancer, pancreatic cancer, biliary tract cancer, colon cancer, rectal cancer, liver cancer, kidney cancer, lung cancer, testicular cancer, breast cancer, ovarian cancer, pancreatic cancer, brain cancer, and head and neck cancer, melanoma, sarcoma, multiple myeloma, leukemia, lymphoma, and the like.

[0114] In one embodiment, the cancer is a hematological cancer or a solid tumor.

[0115] Cancers that develop in blood-forming tissues (e.g., bone marrow) or cells of the immune system are called hematological or blood cancers. Hematological cancers affect the production and function of blood cells and are classified into three major types: leukemia, lymphoma, and multiple myeloma.

[0116] As used herein, "leukemia" refers to a cancer of the blood caused by the rapid production of abnormal white blood cells. Examples of leukemia include: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.

[0117] As used herein, "lymphoma" refers to a type of blood cancer that affects the lymphatic system. Examples of lymphomas include: AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sézary syndrome, cutaneous T-cell lymphoma, and Waldenstrom's macroglobulinemia.

[0118] As used herein, "myeloma" is a cancer of plasma cells. Examples of myeloma include: chronic myeloproliferative neoplasm, Langerhans cell histiocytosis, multiple myeloma, plasma cell neoplasm, myelodysplastic syndrome, and myelodysplastic / myeloproliferative neoplasm.

[0119] In some embodiments, the anti-CCR8 antibodies of the present invention may be used alone or in combination with any other suitable compound known to be capable of treating the disease or indication.

[0120] In some embodiments, administration may be combined with one or more additional therapeutic agents. Phrases such as "combination therapy," "combined with," and the like refer to the simultaneous use of multiple drugs or treatments to enhance efficacy. The compositions of the present invention may be used in combination with other drugs or treatments used, for example, in the treatment of cancer. Specifically, administration of the compositions of the present invention to a subject may be combined with any anti-cancer therapy. Such therapy may be administered before, simultaneously with, or after administration of the compositions of the present invention.

[0121] In some embodiments, the anti-CCR8 antibody is administered to a subject (e.g., a subject with cancer) simultaneously with or sequentially with another treatment. For example, the anti-CCR8 antibody can be administered in combination with one or more of the following: radiation therapy, surgery, or chemotherapy (e.g., targeted chemotherapy or immunotherapy). The start of administration of both agents can be, for example, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 3 days, 5 days, 7 days, or one week or more apart. Alternatively, administration of the second agent can be initiated, for example, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 3 days, 5 days, 7 days, or one week or more after administration of the first agent.

[0122] In some embodiments, the hematological cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sezary syndrome, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma.

[0123] Examples of solid cancers include, but are not limited to, carcinoma, sarcoma, squamous cell carcinoma, small cell lung cancer, pituitary cancer, esophageal cancer, astrocytoma, soft tissue sarcoma, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver tumor, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, renal cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic carcinoma, brain cancer, endometrial cancer, testicular cancer, bile duct cancer, gallbladder cancer, gastric cancer, melanoma, and various types of head and neck cancer.

[0124] In other embodiments, the solid tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal carcinoma, ovarian cancer, liver cancer, or prostate cancer.

[0125] In another embodiment, the present invention provides a method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of an isolated monoclonal antibody or antigen-binding fragment thereof having: HCVR including: a CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15), thereby treating cancer in said subject.

[0126] In some embodiments, the methods described herein further comprise administering to the patient a chemotherapeutic agent, an anti-angiogenic agent, an anti-proliferative agent, an immuno-oncology agent, and / or an anti-tumor composition.

[0127] A "chemotherapeutic agent" is a compound that may be useful in the treatment of cancer. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents (e.g., thiotepa and cyclophosphamide); alkylsulfonates (e.g., busulfan, improsulfan, and piposulfan); aziridines (e.g., benzodopa, carboquone, meturedopa, and uredopa); ethyleneimines and methylamelamines (including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine); acetogenins (particularly bullatacin and bullatacinone); camptothecin (including the synthetic analog topotecan); bryostatin; kallistatin; CC-1065 (including its adozelesin, carzelesin, and bizelesin synthetic analogs); cryptoxanthan gum; ficins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including the synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards (e.g., chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicin (novembicin) ichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas (e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine); antibiotics (e.g., enediyne antibiotics (e.g., calicheamicins, particularly calicheamicin gamma 2 (gammall) and calicheamicin omega 2 (omegall) (see, e.g., Agnew, Chem Intl. Ed. Engl, 33: 183-186 (1994)); dynemicins (including dynemicin A); bisphosphonates (e.g., clodronate); esperamicin;and neocarzinostatin chromophore and related chromoproteins (enediyne antibiotic chromophores); aclacinomycins; actinomycin; anthramycin; azaserine; bleomycin; cactinomycin; carabicin; carminomycin; carzinophilin; chromomycinis; dactinomycin; daunorubicin; detorubicin; 6-diazo-5-oxo-L-norleucine; doxorubicin (including morpholinodoxorubicin, cyanomorpholinodoxorubicin, 2-pyrrolinodoxorubicin, and deoxydoxorubicin); epirubicin; esorubicin; idarubicin; marcellomycin; mitomycins (e.g., mitomycin C); mycophenolic acid; nogalamycin; olivomycin; peplomycin; porfiromycin; purinomycin thromycin; quelamycin; lodorubicin; streptonigrin; streptozocin; tubercidin; ubenimex; zinostatin; zorubicin; antimetabolites (e.g., methotrexate and 5-fluorouracil (5-FU)); folic acid analogs (e.g., denopterin, methotrexate, pteropterin, trimetrexate); purine analogs (e.g., fludarabine, 6-mercaptopurine, thiamiprine, thioguanine) ); pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine); androgens (e.g., calsterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone); antiadrenal agents (e.g., aminoglutethimide, mitotane, trilostane); folic acid supplements (e.g., folinic acid); aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatrexate; defofamine; demecolcine; diaziconazole; eflornithine; elliptinium acetate;Epothilones; etoglucides; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids (e.g., maytansine and ansamitocins); mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; polysaccharide complex (JHS Natural Products, Eugene, Oregon); razoxane; rhizoxin; schizofiran; spirogermanium; tenuazonic acid; triazicones; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verrucarin A) A), Roridin A, and Anguidine); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids (e.g., paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), paclitaxel in a Cremophor-free albumin-modified nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, IL), and docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs (e.g., cisplatin, oxaliplatin, and carboplatin); vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; Xeloda; ibandronate; irinotecan (Camptosar, CPT-11) (including combination regimens of irinotecan with 5-FU and leucovorin); the topoisomerase inhibitor RFS2000;difluoromethylornithine (DFMO); retinoids (e.g., retinoic acid); capecitabine; combretastatin; leucovorin (LV); oxaliplatin (including the oxaliplatin treatment regimen (FOLFOX)); inhibitors of PKC-α, Raf, H-Ras, EGFR (e.g., erlotinib), and VEGF-A that suppress cell proliferation; and pharmaceutically acceptable salts, acids, or derivatives of any of the above;

[0128] Further non-limiting examples of chemotherapeutic agents include: antihormonal agents, which act to regulate or inhibit hormone action on cancer (e.g., antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, ketoxifene, LY117018, onapristone, and toremifene); aromatase inhibitors, which inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands (e.g., 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole); anti-androgens (e.g., flutamide, nilutamide, bicalutamide, leuprorelin, and goserelin); and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, particularly those that inhibit gene expression in signal transduction pathways involved in abnormal cell growth (e.g., PKC-α, Raf, and H-Ras); ribozymes (e.g., VEGF expression inhibitors and HER2 expression inhibitors); vaccines (e.g., gene therapy vaccines); rIL-2; topoisomerase 1 inhibitors; rmRH; and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0129] An "anti-angiogenesis agent" or "angiogenesis inhibitor" refers to a low molecular weight substance, polynucleotide (including, for example, inhibitory RNA (RNAi or siRNA)), polypeptide, isolated protein, recombinant protein, antibody, or conjugate or fusion protein thereof that directly or indirectly inhibits angiogenesis, vasculogenesis, or undesirable vascular permeability. It should be understood that an anti-angiogenesis agent includes an agent that binds to an angiogenic factor or its receptor and blocks its angiogenic activity. For example, anti-angiogenic agents are antibodies or other antagonists to angiogenic agents, such as antibodies to VEGF-A (e.g., bevacizumab) or VEGF-A receptors (e.g., KDR receptors or Flt-1 receptors), anti-PDGFR inhibitors (e.g., imatinib mesylate), small molecules that block VEGF receptor signaling (e.g., PTK787 / ZK2284, SU6668, / SUI 1248 (sunitinib malate), AMG706, or those described, for example, in International Patent Application WO 2004 / 113304). Anti-angiogenic agents also include natural angiogenesis inhibitors (e.g., angiostatin, endostatin, etc.). See, for example, Klagsbrun and D'Amore (Annu. Rev. Physiol. 1991, vol. 53: 217-39); Streit and Detmar (Oncogene 2003, vol. 22: 3172-3179) (e.g., Table 3 listing antiangiogenic therapies in malignant melanoma); Ferrara & Alitalo (Nature Medicine 1999, vol. 5 (12): 1359-1364); Tonini et al. (Oncogene 2003, vol. 22: 6549-6556); and Sato (Int. J. Clin. Oncol. 2003, vol. 8: 200-206; e.g., Table 1 listing antiangiogenic agents used in clinical trials).

[0130] As used herein, the term "growth inhibitory agent" refers to a compound or composition that inhibits the proliferation of cells (e.g., cells expressing VEGF) in vitro or in vivo. Thus, a growth inhibitory agent can significantly reduce the proportion of cells in S phase (e.g., cells expressing VEGF). Examples of growth inhibitory agents include, but are not limited to, agents that block cell cycle progression (at a stage other than S phase) (e.g., agents that induce G1 arrest and M-phase arrest). Classical M-phase blockers include vinca alkaloids (vincas) (vincristine and vinblastine), taxanes, and topoisomerase II inhibitors (e.g., doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin). Agents that arrest the G1 phase also affect S-phase arrest, and examples of such agents include DNA alkylating agents (e.g., tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C). Further information can be found in Chapter 1, "Cell cycle regulation, oncogenes, and antineoplastic drugs," in Mendelsohn and Israel, eds., The Molecular Basis of Cancer, by Murakami et al. (WB Saunders, Philadelphia, 1995), e.g., p. 13. The taxanes (paclitaxel and docetaxel) are anticancer drugs, both derived from the yew tree. Docetaxel (Rhone-Poulenc Rorer) is derived from the European yew tree and is a semisynthetic analog of paclitaxel (Bristol-Myers Squibb). Paclitaxel and docetaxel promote the assembly of microtubules made from tubulin dimers and stabilize microtubules by preventing depolymerization, resulting in the inhibition of cell mitosis.

[0131] The term "anti-neoplastic composition" refers to a composition useful in treating cancer, comprising at least one effective therapeutic agent. Examples of therapeutic agents include, but are not limited to, chemotherapeutic agents, antiproliferative agents, cytotoxic agents, agents used in radiation therapy, anti-angiogenic agents, cancer immunotherapeutic agents (also called immuno-oncology agents), apoptotic agents, anti-tubulin agents, and other agents for treating cancer, such as anti-HER-2 antibodies, anti-CD20 antibodies, epidermal growth factor receptor (EGFR) antagonists (e.g., tyrosine kinase inhibitors), HER1 / EGFR inhibitors (e.g., erlotinib), platelet-derived growth factor inhibitors (e.g., imatinib mesylate), COX-2 inhibitors (e.g., celecoxib), interferons, CTLA4 inhibitors (e.g., For example, anti-CTLA antibody ipilimumab), PD-1 inhibitors (e.g., anti-PD1 antibody BMS-936558), PDL1 inhibitors (e.g., anti-PDL1 antibody MPDL3280A), PDL2 inhibitors (e.g., anti-PDL2 antibodies), VISTA inhibitors (e.g., anti-VISTA antibodies), cytokines, antagonists (e.g., neutralizing antibodies) that bind to one or more of the following targets: ErbB2, ErbB3, ErbB4, PDGFR-β, BlyS, APRIL, BCMA, PD-1, PDL1, PDL2, CTLA4, VISTA, or VEGF receptor(s), TRAIL / Apo2, and other bioactive agents and organic chemical agents. Combinations thereof are also encompassed by the present invention.

[0132] "Checkpoint inhibitor therapy" is a current form of cancer treatment that utilizes immune checkpoints to affect immune system function. Immune checkpoints can be stimulatory or inhibitory. Tumors can use these checkpoints to protect themselves from attack by the immune system. Checkpoint therapy can block inhibitory checkpoints to restore immune system function. Checkpoint proteins include: programmed cell death 1 protein (PDCD1, PD-1; also known as CD279) and its ligand, PD-1 ligand 1 (PD-L1, CD274), cytotoxic T lymphocyte antigen 4 (CTLA-4), A2AR (adenosine A2A receptor), B7-H3 (i.e., CD276), B7-H4 (i.e., VTCN1), BTLA (B and T lymphocyte attenuating factor, i.e., CD272), IDO (indoleamine 2,3-dioxygenase), KIR (killer cell immunoglobulin-like receptor), LAG3 (lymphocyte activation gene-3), TIM-3 (T cell immunoglobulin domain and mucin domain 3), and VISTA (V domain Ig inhibitor of T cell activation).

[0133] Programmed cell death protein 1, also known as PD-1 and CD279 (cluster of differentiation 279), is a cell surface receptor that plays a key role in downregulating the immune system and promoting self-tolerance by suppressing T cell inflammatory activity. PD-1 is an immune checkpoint that protects against autoimmunity through a dual mechanism: it promotes apoptosis (programmed cell death) of antigen-specific T cells in lymph nodes while simultaneously suppressing apoptosis of regulatory T cells (anti-inflammatory, suppressor T cells).

[0134] PD-1 has two ligands, PD-L1 and PD-L2, which belong to the B7 family. PD-L1 protein expression is upregulated in macrophages and dendritic cells (DCs) in response to LPS and GM-CSF treatment, and in T cells and B cells by TCR and B cell receptor signaling. In resting mice, PD-L1 mRNA can be detected in the heart, lung, thymus, spleen, and kidney. PD-L1 is expressed in almost all mouse tumor cell lines, including PA1 myeloma, P815 mastocytoma, and B16 melanoma, upon IFN-γ treatment. PD-L2 expression is more restricted, primarily in DCs and a few tumor cell lines.

[0135] CTLA4 or CTLA-4 (cytotoxic T lymphocyte antigen 4), also known as CD152 (cluster of differentiation 152), is a protein receptor that functions as an immune checkpoint and downregulates immune responses. CTLA4 is constitutively expressed in regulatory T cells but is upregulated in normal T cells only after activation, a phenomenon particularly prominent in cancer. CTLA4 belongs to the immunoglobulin superfamily and is expressed by activated T cells, transmitting inhibitory signals to T cells. CTLA4 shares homology with the T cell costimulatory protein CD28, and both molecules bind to CD80 and CD86 (also known as B7-1 and B7-2, respectively) on antigen-presenting cells. CTLA-4 binds to CD80 and CD86 with higher affinity and avidity than CD28, thereby outcompeting CD28 for binding to their ligands. CTLA-4 transmits inhibitory signals to T cells, while CD28 transmits stimulatory signals. CTLA-4 is also found on regulatory T cells, where it contributes to their suppressive function. T cell activation via the T cell receptor and CD28 increases the expression of CTLA-4.

[0136] There are several checkpoint inhibitors currently being used to treat cancer. PD-1 inhibitors include pembrolizumab (Keytruda) and nivolumab (Opdivo). PD-L1 inhibitors include atezolizumab (Tecentriq), avelumab (Bavencio), and durvalumab (Imfinzi). CTLA-4 inhibitors include ipilimumab (Yervoy). There are also several other checkpoint inhibitors in development, including an anti-B7-H3 antibody (MGA271), an anti-KIR antibody (lirilumab), and an anti-LAG3 antibody (BMS-986016).

[0137] In additional embodiments, the invention provides polynucleotides encoding the heavy or light chain, or antigen-binding portion thereof, of any one of the antibodies or antigen-binding fragments thereof described herein.

[0138] The present invention also provides nucleic acid molecules comprising polynucleotides encoding one or more chains of an antibody (e.g., an anti-CCR8 antibody) described herein. In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a heavy chain or a light chain of an antibody described herein. In some embodiments, the nucleic acid molecule comprises both a polynucleotide encoding a heavy chain and a polynucleotide encoding a light chain of an antibody described herein. In some embodiments, a first nucleic acid molecule comprises a first polynucleotide encoding a heavy chain, and a second nucleic acid molecule comprises a second polynucleotide encoding a light chain.

[0139] In some such embodiments, the heavy and light chains are expressed from one nucleic acid molecule or as two separate polypeptides from two separate nucleic acid molecules, in some embodiments, for example when the antibody is an scFv, a single polynucleotide encodes a single polypeptide in which both the heavy and light chains are linked.

[0140] In some embodiments, a polynucleotide encoding a heavy or light chain of an antibody described herein comprises a nucleotide sequence encoding a leader sequence, which is located at the N-terminus of the heavy or light chain after translation. As noted above, the leader sequence may be the native heavy or light chain leader sequence or may be another exogenous leader sequence.

[0141] As used herein, the terms "nucleic acid" or "oligonucleotide" refer to polynucleotides such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Nucleic acids include, but are not limited to, genomic DNA, cDNA, mRNA, iRNA, miRNA, tRNA, ncRNA, rRNA, and recombinantly produced and chemically synthesized molecules, such as aptamers, plasmids, antisense DNA strands, shRNA, ribozymes, conjugated nucleic acids, and oligonucleotides. According to the present invention, nucleic acids can exist as single-stranded or double-stranded molecules, and as linear or covalently closed circular molecules. Nucleic acids can be isolated. The term "isolated nucleic acid" means that the nucleic acid has been (i) amplified in vitro, e.g., via polymerase chain reaction (PCR), (ii) recombinantly produced by cloning, (iii) purified, e.g., by cleavage and separation by gel electrophoresis, (iv) synthesized, e.g., by chemical synthesis, or (v) extracted from a sample. Nucleic acids can be used for introduction into cells (i.e., cell transfection), particularly in the form of RNA, which can be prepared by in vitro transcription from a DNA template. The RNA can be further modified by stabilizing sequences, capping, and polyadenylation before application.

[0142] In one embodiment, the present invention provides a vector comprising a polynucleotide described herein, The vector is an expression vector selected from the group consisting of a mammalian expression vector, a yeast expression vector, an insect expression vector, and a bacterial expression vector. A vector is provided.

[0143] Vectors comprising polynucleotides encoding the heavy and / or light chains of the antibodies described herein are provided. Such vectors include, but are not limited to, DNA vectors, phage vectors, viral vectors, retroviral vectors, etc. In some embodiments, the vector comprises a first polynucleotide sequence encoding the heavy chain and a second polynucleotide sequence encoding the light chain. In some embodiments, the heavy and light chains are expressed as two separate polypeptides from the vector. In some embodiments, the heavy and light chains are expressed as part of a single polypeptide, for example, when the antibody is an scFv.

[0144] In some embodiments, a first vector comprises a polynucleotide encoding a heavy chain, and a second vector comprises a polynucleotide encoding a light chain. In some embodiments, the first vector and the second vector are transfected into host cells in similar amounts (e.g., similar molar amounts or similar mass amounts). In some embodiments, the first vector and the second vector are transfected into host cells in a molar or mass ratio of between 5:1 and 1:5. In some embodiments, a mass ratio of between 1:1 and 1:5 is used for the vector encoding the heavy chain and the vector encoding the light chain. In some embodiments, a mass ratio of 1:2 is used for the vector encoding the heavy chain and the vector encoding the light chain.

[0145] In some embodiments, a vector optimized for the expression of a polypeptide in CHO cells or CHO-derived cells, or in NSO cells, is selected. Examples of such vectors are described, for example, in Running Deer et al., Biotechnol. Prog. 20: 880-889 (2004). In some embodiments, a vector is selected for the in vivo expression of a CCR8 antagonist in animals (including humans). In some such embodiments, the expression of a polypeptide(s) is under the control of a promoter(s) that functions in a tissue-specific manner. For example, a liver-specific promoter is described, for example, in PCT Publication No. WO 2006 / 076288.

[0146] The terms "vector," "expression vector," or "plasmid DNA," as used herein, refer to a recombinant nucleic acid construct engineered by human intervention. A recombinant nucleic acid construct may contain two or more nucleotide sequences linked in a manner whose products are not naturally found in a cell. Specifically, two or more nucleotide sequences may be operably linked, for example, a gene encoding a protein of interest, one or more protein tags, a functional domain, etc.

[0147] Vectors suitable for use in preparing proteins and / or protein conjugates include those selected from the following: baculovirus, phage, plasmid, phagemid, cosmid, fosmid, bacterial artificial chromosome, viral DNA, P1-based artificial chromosome, yeast plasmid, and yeast artificial chromosome. For example, viral DNA vectors can be selected from the following: vaccinia virus, adenovirus, foul pox virus, pseudorabies virus, and SV40 derivatives. One type of vector is a genome-integrating vector, or "integrated vector," which can integrate into the chromosomal DNA of a host cell. Another type of vector is an episomal vector, e.g., a nucleic acid capable of extrachromosomal replication. A vector capable of expressing a gene when operably linked to it is referred to herein as an "expression vector." Viral vectors include adenovirus, adeno-associated virus (AAV), retrovirus, lentivirus, vaccinia virus, measles virus, herpes virus, and bovine papillomavirus vectors (for a review of viral and non-viral vectors, see Kay et al., Proc. Natl. Acad. Sci. USA 94:12744-12746 (1997)). Viral vectors are modified to alter or eliminate the virus's natural tropism and pathogenicity. The viral genome can also be modified to enhance its infectivity and accommodate packaging of a nucleic acid encoding a polypeptide of interest.

[0148] The nucleic acid constructs of the present invention can be introduced into cells to modify the cells and enable expression of chimeric proteins in the cells. Various suitable methods for introducing nucleic acids into cells are known in the art and include viral and non-viral techniques. Typical non-viral techniques include, but are not limited to, electroporation, calcium phosphate-mediated introduction, nucleofection, sonoporation, heat shock, magnetofection, liposome-mediated introduction, microinjection, microprojectile-mediated introduction (nanoparticles), cationic polymer-mediated introduction (DEAE-dextran, polyethyleneimine, polyethylene glycol (PEG), etc.), or cell fusion. Other transfection methods include, for example, the following proprietary transfection reagents: Lipofectamine™, Dojindo Hilymax™, Fugene™, jetPEI™, Effectene™, and DreamFect™.

[0149] The nucleic acid constructs of the present invention can be introduced into host cells to modify the cells and enable expression of chimeric proteins in the cells. Various host cells suitable for expressing chimeric proteins are known in the art. Typical examples of cells used for transfection include, but are not limited to, bacterial cells, eukaryotic cells, yeast cells, insect cells, or plant cells, such as E. coli, Bacillus, Streptomyces, Pichia pastoris, Salmonella typhimurium, Drosophila S2, Spodoptera SJ9, CHO, COS (e.g., COS-7), 3T3-F442A, HeLa, HUVEC, HUAEC, NIH 3T3, Jurkat, 293, 293H, or 293F.

[0150] In various embodiments, the heavy and / or light chains of the antibodies described herein can be expressed in prokaryotic cells, such as bacterial cells; or in eukaryotic cells, such as fungal cells (e.g., yeast), plant cells, insect cells, and mammalian cells. Such expression can be carried out, for example, according to procedures known in the art. Examples of eukaryotic cells that can be used to express polypeptides include, but are not limited to, COS cells (including COS-7 cells); 293 cells (including 293-6E cells); CHO cells (including CHO-S and DG44 cells); PER.C6® cells (Crucell); and NSO cells. In some embodiments, the heavy and / or light chains of the antibodies described herein can be expressed in yeast. See, e.g., U.S. Patent Publication No. US 2006 / 0270045 A1. In some embodiments, a particular eukaryotic host cell is selected based on its ability to make desired post-translational modifications to the heavy and / or light chains of the anti-CCR8 antibody. For example, in some embodiments, CHO cells produce polypeptides with higher sialylation levels compared to the same polypeptides produced in 293 cells.

[0151] In various embodiments herein, the anti-CCR8 monoclonal antibodies of the invention are expressed in cells with limited or no fucosylation, resulting in an antibody with increased defucosylation. In some embodiments, the antibody is selected from: (a) an antibody having a heavy chain comprising SEQ ID NO: 3 and a light chain comprising SEQ ID NO: 2; (b) an antibody having a heavy chain comprising SEQ ID NO: 4 and a light chain comprising SEQ ID NO: 2; (c) an antibody having a heavy chain comprising SEQ ID NO: 5 and a light chain comprising SEQ ID NO: 2; (d) an antibody having a heavy chain comprising SEQ ID NO: 6 and a light chain comprising SEQ ID NO: 2; (e) an antibody having a heavy chain comprising SEQ ID NO: 7 and a light chain comprising SEQ ID NO: 2; (f) an antibody having a heavy chain comprising SEQ ID NO: 8 and a light chain comprising SEQ ID NO: 2; (h) an antibody having a heavy chain comprising SEQ ID NO: 9 and a light chain comprising SEQ ID NO: 2; The antibody specifically binds to CCR8. In preferred embodiments, the antibody has a heavy chain comprising SEQ ID NO:6 and a light chain comprising SEQ ID NO:2. In these embodiments, the antibody has a defucosylation level of at least 50%. For example, the antibody has a defucosylation level of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more.

[0152] Introduction of one or more nucleic acids into desired host cells can be achieved by any method, including, but not limited to, calcium phosphate transfection, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, etc. Non-limiting examples of methods are described, for example, in Sambrook et al., Molecular Cloning, A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press (2001). Nucleic acids can be transiently or stably transfected into desired host cells according to any suitable method.

[0153] In some embodiments, one or more polypeptides may be produced in vivo in an animal that has been modified or transfected with one or more nucleic acid molecules encoding the polypeptides, according to any suitable method.

[0154] Polynucleotides can be delivered to cells (e.g., multiple different cells or cell types, including target cells or cell types and / or non-target cell types) using vectors (e.g., expression vectors). Examples of vectors include, but are not limited to: (a) non-viral vectors (e.g., nucleic acid vectors, including linear oligonucleotides and circular plasmids; artificial chromosomes, such as human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), and bacterial artificial chromosomes (BACs or PACs); episomal vectors; transposons (e.g., PiggyBacs)); and (b) viral vectors (e.g., retroviral vectors, lentiviral vectors, adenoviral vectors, and AAV vectors). Viral vectors have several advantages for nucleic acid delivery, including high infectivity and / or tropism for specific target cells or tissues. In some cases, viral vectors can be used to deliver the polynucleotides described herein.

[0155] [Additional Embodiments] Embodiment 1. An isolated monoclonal antibody or antigen-binding fragment thereof comprising: HCVR including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15), the antibody or antigen-binding fragment thereof specifically binds to CCR8; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0156] Embodiment 2. An isolated monoclonal antibody or antigen-binding fragment thereof described in embodiment 1, further comprising the HFR1 sequence of SEQ ID NO: 23, the HFR2 sequence of SEQ ID NO: 24, the HFR3 sequence of SEQ ID NO: 25, and / or the HFR4 sequence of SEQ ID NO: 26; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0157] Embodiment 3. An isolated monoclonal antibody or antigen-binding fragment thereof according to embodiment 1 or 2, further comprising an LFR1 sequence of SEQ ID NO: 27, an LFR2 sequence of SEQ ID NO: 28, an LFR3 sequence of SEQ ID NO: 29, and / or an LFR4 sequence of SEQ ID NO: 30; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0158] Embodiment 4. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 3, the HCVR sequence is SEQ ID NO: 36 and / or the LCVR sequence is SEQ ID NO: 32; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0159] Embodiment 5. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, The HCVR sequence is SEQ ID NO: 36, and the LCVR sequence is SEQ ID NO: 32. An isolated monoclonal antibody or antigen-binding fragment thereof.

[0160] Embodiment 6. An isolated monoclonal antibody according to any one of embodiments 1 to 5, The heavy chain sequence is SEQ ID NO: 6 and the light chain sequence is SEQ ID NO: 2. Isolated monoclonal antibodies.

[0161] Embodiment 7. An isolated monoclonal antibody or antigen-binding fragment thereof, comprising: (a) HCVR, which includes: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSEQNYYYAMDY (SEQ ID NO: 16), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (b) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENNYYYAMDY (SEQ ID NO: 17), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (c) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSENQYYYAMDY (SEQ ID NO: 18), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (d) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDQNYYYAMDY (SEQ ID NO: 20), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (e) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDNQYYYAMDY (SEQ ID NO: 21), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (f) HCVR, including: the HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); an HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11), and an HCVR CDR3 sequence having the amino acid sequence TRGSDQQYYYAMDY (SEQ ID NO: 22), and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13), an LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14), and an LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); the antibody or antigen-binding fragment thereof specifically binds to CCR8; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0162] Embodiment 8. An isolated monoclonal antibody or antigen-binding fragment thereof according to embodiment 7, further comprising the HFR1 sequence of SEQ ID NO: 23, the HFR2 sequence of SEQ ID NO: 24, the HFR3 sequence of SEQ ID NO: 25, and / or the HFR4 sequence of SEQ ID NO: 26; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0163] Embodiment 9. An isolated monoclonal antibody or antigen-binding fragment thereof according to embodiment 7 or 8, further comprising the LFR1 sequence of SEQ ID NO: 27, the LFR2 sequence of SEQ ID NO: 28, the LFR3 sequence of SEQ ID NO: 28, and / or the LFR4 sequence of SEQ ID NO: 29; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0164] Embodiment 10. An isolated monoclonal antibody or antigen-binding fragment thereof, comprising: (a) an HCVR sequence of SEQ ID NO: 33 and an LCVR sequence of SEQ ID NO: 32; or (b) an HCVR sequence of SEQ ID NO: 34 and an LCVR sequence of SEQ ID NO: 32; or (c) an HCVR sequence of SEQ ID NO: 35 and an LCVR sequence of SEQ ID NO: 32; or (d) an HCVR sequence of SEQ ID NO: 37 and an LCVR sequence of SEQ ID NO: 32; or (e) an HCVR sequence of SEQ ID NO: 38 and an LCVR sequence of SEQ ID NO: 32; or (f) an HCVR sequence of SEQ ID NO: 39 and an LCVR sequence of SEQ ID NO: 32; the antibody or antigen-binding fragment thereof specifically binds to CCR8; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0165] Embodiment 11. An isolated monoclonal antibody, comprising: (a) a heavy chain sequence of SEQ ID NO: 3 and a light chain sequence of SEQ ID NO: 2; or (b) a heavy chain sequence of SEQ ID NO: 4 and a light chain sequence of SEQ ID NO: 2; or (c) a heavy chain sequence of SEQ ID NO: 5 and a light chain sequence of SEQ ID NO: 2; or (d) a heavy chain sequence of SEQ ID NO: 7 and a light chain sequence of SEQ ID NO: 2; or (e) a heavy chain sequence of SEQ ID NO: 8 and a light chain sequence of SEQ ID NO: 2; or (f) a heavy chain sequence of SEQ ID NO: 9 and a light chain sequence of SEQ ID NO: 2; the antibody or antigen-binding fragment thereof specifically binds to CCR8; Isolated monoclonal antibodies.

[0166] Embodiment 12. An isolated monoclonal antibody or antigen-binding fragment thereof having at least 80% identity to the antibody of any one of embodiments 5, 6, 10, or 11, the antibody or antigen-binding fragment thereof specifically binds to CCR8; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0167] Embodiment 13. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 12, The antibody is a humanized antibody. An isolated monoclonal antibody or antigen-binding fragment thereof.

[0168] Embodiment 14. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 13, the antigen-binding fragment thereof is a Fab, Fab', F(ab')2, Fd, single-chain Fv or scFv, disulfide-linked Fv, V-NAR domain, IgNar, intrabody, IgGACH2, minibody, F(ab')3, tetrabody, triabody, diabody, domain antibody (dAb), DVD-Ig, Fcab, mAb2, (scFv)2, tandem scFv, DART®, TandAb, nanobody, or scFv-Fc; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0169] Embodiment 15. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 14, the monoclonal antibody or antigen-binding fragment thereof binds to human CCR8 with a Kd of less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, 1 nM, 0.5 nM, or 0.01 nM; An isolated monoclonal antibody or antigen-binding fragment thereof.

[0170] Embodiment 16. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 15, Induce ADCC, An isolated monoclonal antibody or antigen-binding fragment thereof.

[0171] Embodiment 17. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 16, The antibody has a defucosylation level of at least 50%. An isolated monoclonal antibody or antigen-binding fragment thereof.

[0172] Embodiment 18. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 17, The antibody has a defucosylation level of at least 85%. An isolated monoclonal antibody or antigen-binding fragment thereof.

[0173] Embodiment 19. A method of treating cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-18, thereby treating cancer in the subject. method.

[0174] Embodiment 20. A method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-18 and an anti-cancer agent, thereby treating cancer in the subject. method.

[0175] Embodiment 21. The method of embodiment 19 or 20, further comprising: the cancer is a blood cancer or a solid tumor; method.

[0176] Embodiment 22. The method of embodiment 21, further comprising: The hematological cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sezary syndrome, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma. method.

[0177] Embodiment 23. The method of embodiment 21, further comprising: The solid tumor is breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, or prostate cancer; method.

[0178] Embodiment 24. The method of embodiment 19, further comprising: The method further comprises administering to the patient an anti-cancer agent. method.

[0179] Embodiment 25. The method of embodiment 20 or 24, further comprising: The anti-cancer agent is a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition; method.

[0180] Embodiment 26. A method of treating breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, and / or prostate cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-18, thereby treating cancer in the subject. method.

[0181] Embodiment 27. A method of treating breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, and / or prostate cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-18 and an anti-cancer agent, thereby treating cancer in the subject. method.

[0182] Embodiment 28. The method of embodiment 27, further comprising: The anti-cancer agent is a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition; method.

[0183] Embodiment 29. A method of treating cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-6, thereby treating cancer in the subject. method.

[0184] Embodiment 30. A method of treating breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, and / or prostate cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-6, thereby treating cancer in the subject. method.

[0185] Embodiment 31. A method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-6 and an anti-cancer agent, thereby treating cancer in the subject. method.

[0186] Embodiment 32. A method of treating breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, and / or prostate cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-6 and an anti-cancer agent, thereby treating cancer in the subject. method.

[0187] Embodiment 33 The method of embodiment 29 or 31, comprising: the cancer is a blood cancer or a solid tumor; method.

[0188] Embodiment 34. The method of embodiment 33, further comprising: The hematological cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sezary syndrome, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma. method.

[0189] Embodiment 35. The method of embodiment 33, further comprising: The solid tumor is breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, or prostate cancer; method.

[0190] Embodiment 36 The method of embodiment 29 or 30, further comprising: The method further comprises administering to the patient an anti-cancer agent. method.

[0191] Embodiment 37. The method of embodiment 36, further comprising: The anti-cancer agent is a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition; method.

[0192] Embodiment 38. A polynucleotide encoding the heavy or light chain, or an antigen-binding portion thereof, of any one of Embodiments 1 to 18.

[0193] Embodiment 39. A vector comprising the polynucleotide of embodiment 38, The vector is an expression vector selected from the group consisting of a mammalian expression vector, a yeast expression vector, an insect expression vector, and a bacterial expression vector. vector.

[0194] Embodiment 40. A method for producing a monoclonal antibody according to embodiment 6 or 11, comprising: the antibody is at least 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 99% defucosylated; The method includes expressing a polynucleotide encoding the antibody in a cell that has been modified to reduce or prevent fucosylation of the antibody. method.

[0195] The following are examples relating to the isolated anti-CCR8 antibodies described herein. The following examples are provided to further illustrate embodiments of the present invention, but are not intended to limit the scope of the invention. While they are typical of those that may be used, other procedures, methodologies, or techniques known to those skilled in the art may be used instead. [Example]

[0196] Example 1: Defucosylation of humanized anti-CCR8 antibody To assess the impact of defucosylation level on the properties of the Ab001_WT antibody, ATUM's proprietary hypofucosylation technology (miFuc™) was applied during cell line generation to increase the defucosylation level to >90%. A second control cell line with the same sequence but without the miFuc™ hypofucosylation technology achieved approximately 11% defucosylation. Antibody efficacy via ADCC was assessed using a HuT78 cell reporter assay using Jurkat-CD16A-NFAT. An approximately 100-fold increase in ADCC activity was observed in the hypofucosylated cell line compared to the non-hypofucosylated cell line (see Table 1 and Figure 1).

[0197] JPEG2025527535000002.jpg44166

[0198] Example 2: CCR8 binding assay Antibodies were tested for binding to CCR8 using the CHO-K1 human chemokine CCR8 receptor cell line (CHO.hCCR8; Perkin Elmer Corp., Waltham, MA) as follows: approximately 5 x 10 4CHO.hCCR8 cells were resuspended in 50 μL of serially diluted antibody solution in cold MACS buffer (Miltenyi Biotec, Inc., San Jose, CA) to antibody concentrations of 10, 3.3, 1.1, 0.37, 0.12, 0.04, and 0.01 nM in each well of a V-bottom 96-well plate. After 30 minutes of incubation on ice, 150 μL of cold MACS buffer was added to each well, the plate was centrifuged at 500 × g for 5 minutes at 4°C, and the supernatant was discarded. The cells were washed twice in 200 μL of cold MACS buffer and then resuspended in 50 μL of anti-human IgG-Alexa Fluor 488 (The Jackson Laboratory, Bar Harbor, ME) diluted 1:100 in MACS buffer. After 30 minutes of incubation on ice, 150 μL of cold MACS buffer was added to each well, the plate was centrifuged at 500 × g for 5 minutes at 4°C, and the supernatant was discarded. Cells were washed twice in 200 μL of cold MACS buffer and then resuspended in 80 μL of cold MACS buffer containing propidium iodide (PI, 1:1000 dilution, 1 μg / ml; ThermoFisher Scientific, Waltham, MA). Antibody binding (luminescence) was measured using a CellStream flow cytometry system (Cytek Biosciences, Fremont, CA).

[0199] Example 3: Antibody-dependent cellular cytotoxicity (ADCC) assay CHO.hCCR8 cells were 1 × 10 4Cells were seeded at a density of 100 μL of Ham's F-12 medium (10% FBS, 0.4 mg / mL G418) into a white, clear-bottom 96-well plate at 100 μL per well. After 24 hours, the medium was removed and 40 μL of assay medium (One-Step Luciferase Assay System; BPS Bioscience, San Diego, CA) was added. Antibodies were diluted 1:10 and then serially diluted 1:4 in assay medium. 10 μL of each was dispensed into wells and incubated for 1 hour. ADCC bioassay effector cell V variant (high affinity)-Jurkat recombinant cell line (BPS Bioscience) cells were added at 5 × 10 per well. 4 50 μL of cells / well was added. The plate was incubated at 37°C and 5% CO for 5 hours, after which 100 μL of luciferase assay working solution (luciferase reagent substrate diluted 1:100 in luciferase reagent buffer; BPS Bioscience) was added, and the plate was gently rocked for 20 minutes at room temperature. Luminescence was then measured using a luminometer.

[0200] [Example 4: Deamination of Ab001_Wt antibody] Ab001_Wt was incubated in PBS (pH 7.4) at 37°C, and samples were taken on days 0, 1, 4, 7, 14, 21, and 28. The samples were analyzed using human CCR8-binding EC 50 , ADCC EC 50 The deamination rates at N104 and / or N105 were assessed at each time point. As shown in Figures 2A and 2B, hCCR8 binding and ADCC activity decreased over time. As shown in Figure 2C, this decrease was consistent with the deamination of Ab001_Wt antibody over the same period. This indicates that Ab001_Wt exhibits the disadvantage of reducing antibody efficacy over time and that frequent administration of the antibody is necessary to maintain the activity level of Ab001_Wt.

[0201] In a second experiment, plasma samples from a monkey pharmacokinetic (PK) study showed that both CCR8 binding (Figure 3A) and ADCC activity (Figure 3B) rapidly decreased over time, indicating a similar disadvantage of Ab001_Wt in animals.

[0202] Example 5: Construction and testing of mutant antibodies Ab001_Wt was modified in the HCVR CDR3 to generate a series of antibodies with reduced susceptibility to deamination. These antibodies modify amino acids 103-105 of the HCVR CDR3 sequence from DNN in the parent antibody Ab001_Wt to ​​EQN in Ab001_M1, ENN in Ab001_M2, ENQ in Ab001_M3, EQQ in Ab001_M4, DQN in Ab001_M5, DNQ in Ab001_M6, and DQQ in Ab001_M7. The mutant antibodies were tested for CCR8 binding affinity and ADCC activity compared to the parent antibody Ab001_Wt.

[0203] As shown in Figures 4A and 4B, the CCR8 binding of Ab001-M1, M2, M3, and M4 was measured and compared with the binding activity of Ab001_Wt. The EC50 values ​​are summarized in Figure 4C. Modifications of the CDR3 region significantly improved the binding EC50 values. 50 Although Ab001_M4 showed some effect on CCR8 binding activity, unexpectedly, complete substitution of asparaginyl residues N104 and N105 in Ab001_M4 did not result in significant changes. Thus, Ab001_M4 overcomes the disadvantage of deamination over time and maintains CCR8 binding activity.

[0204] As shown in Figures 5A and 5B, the ADCC activities of Ab001-M1, M2, M3, and M4 were measured and compared with that of Ab001_Wt. The EC50s are summarized in Figure 5C. Similar to CCR8 binding, modifications in the CDR3 region significantly improved the ADCC EC50s. 50Although Ab001_M4 showed some effect on ADCC activity, unexpectedly, complete substitution of asparaginyl residues N104 and N105 in Ab001_M4 did not result in significant changes. Thus, Ab001_M4 overcomes the disadvantage of deamination over time and maintains ADCC activity.

[0205] Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.

[0206] [Sequence List:] JPEG2025527535000003.jpg14166

[0207] >Ab001_WT HC (wild type) (SEQ ID NO: 1) JPEG2025527535000004.jpg62166

[0208] > Ab001_WT LC (wild type and all variants below) (SEQ ID NO: 2) METDTLLLWVLLLWVPGSTG DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGNTYLYWFLQKPGQSPQLLIYRMSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYYCMQHLEYPFTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0209] > Ab001_M1 HC(EQN)(SEQ ID NO:3) JPEG2025527535000005.jpg63166

[0210] > Ab001_M2 HC(ENN)(SEQ ID NO: 4) JPEG2025527535000006.jpg63166

[0211] > Ab001_M3 HC(ENQ) (SEQ ID NO: 5) JPEG2025527535000007.jpg62166

[0212] > Ab001_M4 HC(EQQ)(SEQ ID NO:6) JPEG2025527535000008.jpg62166

[0213] > Ab001_M5 HC(DQN)(SEQ ID NO:7) JPEG2025527535000009.jpg62166

[0214] > Ab001_M6 HC(DNQ) (SEQ ID NO: 8) JPEG2025527535000010.jpg62166

[0215] > Ab001_M7 HC(DQQ)(SEQ ID NO:9) JPEG2025527535000011.jpg62166

[0216] >Ab001_WT and M1-M7 HCDR1 (SEQ ID NO: 10) GFRFNTNA

[0217] >Ab001_WT and M1-M7 HCDR2 (SEQ ID NO: 11) IRSKSNSYATYY

[0218] >Ab001_WT HCDR3 (SEQ ID NO: 12) TRGSDNNYYYAMDY

[0219] >Ab001_WT and M1-M7 LCDR1 (SEQ ID NO: 13) QSLLHSNGNTY

[0220] >Ab001_WT and M1-M7 LCDR2 (SEQ ID NO: 14) YRMSNR

[0221] >Ab001_WT and M1-M7 LCDR3 (SEQ ID NO: 15) MQHLEYPFT

[0222] >Ab001_M1 HCDR3 (SEQ ID NO: 16) TRGSEQNYYYAMDY

[0223] >Ab001 M2 HCDR3 (SEQ ID NO: 17) TRGSENNYYYAMDY

[0224] >Ab001 M3 HCDR3 (SEQ ID NO: 18) TRGSENQYYYAMDY

[0225] >Ab001 M4 HCDR3 (SEQ ID NO: 19) TRGSEQQYYYAMDY

[0226] >Ab001 M5 HCDR3 (SEQ ID NO: 20) TRGSDQNYYYAMDY

[0227] >Ab001 M6 HCDR3 (SEQ ID NO: 21) TRGSDNQYYYAMDY

[0228] >Ab001 M7 HCDR3 (SEQ ID NO: 22) TRGSDQQYYYAMDY

[0229] >Ab001 WT and M1-M7 HFR1 (SEQ ID NO: 23) EVQLVESGGGLVQPGGSLKLSCAAS

[0230] >Ab001 WT and M1-M7 HFR2 (SEQ ID NO: 24) MNWVRQASGKDLEWVAR

[0231] >Ab001 WT and M1-M7 HFR3 (SEQ ID NO: 25) AASVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYC

[0232] >Ab001 WT and M1-M7 HFR4 (SEQ ID NO: 26) WGQGTTVTVSS

[0233] >Ab001 WT and M1-M7 LFR1 (SEQ ID NO: 27) DIVMTQSPLSLPVTPGEPASISCRSS

[0234] >Ab001 WT and M1-M7 LFR2 (SEQ ID NO: 28) LYWFLQKPGQSPQLLI

[0235] >Ab001 WT and M1-M7 LFR3 (SEQ ID NO: 29) ASGVPDRFSGSGSGTDFTLKISRVEAEEDVGVYYC

[0236] >Ab001 WT and M1-M7 LFR4 (SEQ ID NO: 30) FGQGTKLEIK

[0237] >Ab001_WT HCVR (wild type) (SEQ ID NO: 31) JPEG2025527535000012.jpg21166

[0238] > Ab001_WT LCVR (wild type and all variants below) (SEQ ID NO: 32) JPEG2025527535000013.jpg20166

[0239] > Ab001_M1 HC(EQN) (SEQ ID NO: 33) JPEG2025527535000014.jpg21166

[0240] > Ab001_M2 HC(ENN) (SEQ ID NO: 34) JPEG2025527535000015.jpg22166

[0241] > Ab001_M3 HC(ENQ) (SEQ ID NO: 35) JPEG2025527535000016.jpg21166

[0242] > Ab001_M4 HC(EQQ) (SEQ ID NO: 36) JPEG2025527535000017.jpg21166

[0243] > Ab001_M5 HC(DQN)(SEQ ID NO:37) JPEG2025527535000018.jpg20166

[0244] > Ab001_M6 HC(DNQ) (SEQ ID NO: 38) JPEG2025527535000019.jpg21166

[0245] > Ab001_M7 HC(DQQ) (SEQ ID NO: 39) JPEG2025527535000020.jpg20166

Claims

1. 1. An isolated monoclonal antibody or antigen-binding fragment thereof: A heavy chain variable region (HCVR) comprising: an HCVR complementarity determining region (CDR1) sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); Including, The antigen-binding fragment of the antibody specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof.

2. 1. An isolated monoclonal antibody or antigen-binding fragment thereof: a HC comprising a sequence having at least 80% identity to SEQ ID NO:6 and having the antigen-binding specificity thereof; and LC comprising a sequence having at least 80% identity with SEQ ID NO: 2 and having the antigen-binding specificity thereof. Including, An isolated monoclonal antibody or antigen-binding fragment thereof.

3. 3. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 2, Where: The heavy chain variable region (HCVR) sequence is SEQ ID NO: 36, and / or the light chain variable region (LCVR) sequence is SEQ ID NO: 32; An isolated monoclonal antibody or antigen-binding fragment thereof.

4. 4. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 2 or 3, Where: The HCVR sequence is SEQ ID NO: 36 and the LCVR sequence is SEQ ID NO:

32. An isolated monoclonal antibody or antigen-binding fragment thereof.

5. 10. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 2, 3, or 4, Where: The HC sequence is SEQ ID NO: 6 and the LC sequence is SEQ ID NO:

2. An isolated monoclonal antibody or antigen-binding fragment thereof.

6. 1. An isolated monoclonal antibody or antigen-binding fragment thereof: (a) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSEQNYYYAMDY (SEQ ID NO: 16); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (b) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSENNYYYAMDY (SEQ ID NO: 17); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (c) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSENQYYYAMDY (SEQ ID NO: 18); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (d) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSDQNYYYAMDY (SEQ ID NO: 20); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (e) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSDNQYYYAMDY (SEQ ID NO: 21); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); or (f) a heavy chain variable region (HCVR) comprising: HCVR CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSDQQYYYAMDY (SEQ ID NO: 22); and A light chain variable region (LCVR) comprising: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); Including, The antigen-binding fragment of the antibody specifically binds to CCR8. An isolated monoclonal antibody or antigen-binding fragment thereof.

7. 1. An isolated monoclonal antibody or antigen-binding fragment thereof: (a) a heavy chain (HC) comprising a sequence having at least 80% identity to SEQ ID NO: 3 and having the antigen-binding specificity thereof; and a light chain (LC) comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (b) a HC comprising a sequence having at least 80% identity to SEQ ID NO: 4 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (c) a HC comprising a sequence having at least 80% identity to SEQ ID NO: 5 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (d) a HC comprising a sequence having at least 80% identity to SEQ ID NO: 7 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; (e) a HC comprising a sequence having at least 80% identity to SEQ ID NO: 8 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; or (f) a HC comprising a sequence having at least 80% identity to SEQ ID NO: 9 and having the antigen-binding specificity thereof; and an LC comprising a sequence having at least 80% identity to SEQ ID NO: 2 and having the antigen-binding specificity thereof; Including, the antibody or antigen-binding fragment thereof specifically binds to CCR8; An isolated monoclonal antibody or antigen-binding fragment thereof.

8. 7. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 6, Where: (a) the HCVR sequence is SEQ ID NO: 33 and / or the LCVR sequence is SEQ ID NO: 32; (b) the HCVR sequence is SEQ ID NO: 34 and / or the LCVR sequence is SEQ ID NO: 32; (c) the HCVR sequence is SEQ ID NO: 35 and / or the LCVR sequence is SEQ ID NO: 32; (d) the HCVR sequence is SEQ ID NO: 37 and / or the LCVR sequence is SEQ ID NO: 32; (e) the HCVR sequence is SEQ ID NO: 38 and / or the LCVR sequence is SEQ ID NO: 32; (f) the HCVR sequence is SEQ ID NO: 39 and / or the LCVR sequence is SEQ ID NO: 32; An isolated monoclonal antibody or antigen-binding fragment thereof.

9. 9. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 6 or 8, Where: (a) the HCVR sequence is SEQ ID NO: 33 and the LCVR sequence is SEQ ID NO: 32; (b) the HCVR sequence is SEQ ID NO: 34 and the LCVR sequence is SEQ ID NO: 32; (c) the HCVR sequence is SEQ ID NO: 35 and the LCVR sequence is SEQ ID NO: 32; (d) the HCVR sequence is SEQ ID NO: 37 and the LCVR sequence is SEQ ID NO: 32; (e) the HCVR sequence is SEQ ID NO: 38 and the LCVR sequence is SEQ ID NO: 32; (f) the HCVR sequence is SEQ ID NO: 39 and the LCVR sequence is SEQ ID NO: 32; An isolated monoclonal antibody or antigen-binding fragment thereof.

10. 10. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 6, 8, or 9, Where: The HC sequence is SEQ ID NO: 3 and the LC sequence is SEQ ID NO:

2. The HC sequence is SEQ ID NO: 4 and the LC sequence is SEQ ID NO:

2. The HC sequence is SEQ ID NO: 5 and the LC sequence is SEQ ID NO:

2. The HC sequence is SEQ ID NO: 7 and the LC sequence is SEQ ID NO:

2. The HC sequence is SEQ ID NO: 8 and the LC sequence is SEQ ID NO:

2. or The HC sequence is SEQ ID NO: 9 and the LC sequence is SEQ ID NO:

2. An isolated monoclonal antibody or antigen-binding fragment thereof.

11. 11. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 10, The antibody is a humanized antibody. An isolated monoclonal antibody or antigen-binding fragment thereof.

12. 12. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 11, The antigen-binding fragments thereof include Fab, Fab', F(ab') ２ , F ｄ , single chain Fv or scFv, disulfide bond F ｖ , V-NAR domain, IgNar, intrabody, IgG ACH ２ , minibody, F(ab') ３ , tetrabodies, triabodies, diabodies, single domain antibodies, DVD-Ig, Fcab, mAb ２ , (scFv) ２ or scFv-Fc; An isolated monoclonal antibody or antigen-binding fragment thereof.

13. 13. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 12, The monoclonal antibody or antigen-binding fragment thereof is ｄ binds to human CCR8 with less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, 1 nM, 0.5 nM, or 0.01 nM. An isolated monoclonal antibody or antigen-binding fragment thereof.

14. 14. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 13, induce antibody-dependent cell-mediated cytotoxicity (ADCC); An isolated monoclonal antibody or antigen-binding fragment thereof.

15. 15. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 14, The antibody has a defucosylation level of at least 50%. An isolated monoclonal antibody or antigen-binding fragment thereof.

16. 16. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 15, The antibody has a defucosylation level of at least 85%. An isolated monoclonal antibody or antigen-binding fragment thereof.

17. 1. A method of treating cancer in a subject in need thereof, comprising: The method comprises administering to the subject an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 16, thereby treating cancer in the subject. method.

18. 18. The method of claim 17, the cancer is a blood cancer or a solid tumor; method.

19. 20. The method of claim 18, The hematological cancers are the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma, mycosis fungoides, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Sezary syndrome, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma. Selected from: method.

20. 20. The method of claim 18, The solid tumor is breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, renal cancer, ovarian cancer, liver cancer, or prostate cancer; method.

21. A method according to any one of claims 17 to 20, comprising: The method further comprises administering to the patient a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition. method.

22. 1. A method of treating cancer in a subject in need thereof, comprising: The method includes administering to the subject an effective amount of an isolated monoclonal antibody or antigen-binding fragment thereof; The isolated monoclonal antibody or antigen-binding fragment thereof comprises: HCVR, including: a CDR1 sequence having the amino acid sequence GFRFNTNA (SEQ ID NO: 10); HCVR CDR2 sequence having the amino acid sequence IRSKSNSYATYY (SEQ ID NO: 11); and an HCVR CDR3 sequence having the amino acid sequence TRGSEQQYYYAMDY (SEQ ID NO: 19); and LCVR including: LCVR CDR1 sequence having the amino acid sequence QSLLHSNGNTY (SEQ ID NO: 13); LCVR CDR2 sequence having the amino acid sequence YRMSNR (SEQ ID NO: 14); and LCVR CDR3 sequence having the amino acid sequence MQHLEYPFT (SEQ ID NO: 15); and thereby treating cancer in said subject. method.

23. 23. The method of claim 22, the cancer is a solid tumor; method.

24. 24. The method of claim 23, The solid tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, melanoma, uveal melanoma, colon cancer, kidney cancer, urothelial cancer, ovarian cancer, endometrial cancer, uterine cancer, liver cancer, pancreatic cancer, bile duct cancer, gastric cancer, gastroesophageal cancer, esophageal cancer, cervical cancer, squamous cell carcinoma, prostate cancer, or bladder cancer. Selected from: method.

25. A method according to any one of claims 22 to 24, comprising: The method further comprises administering to the patient a chemotherapeutic agent, an anti-angiogenic agent, a growth inhibitor, an immuno-oncology agent, a checkpoint inhibitor, and / or an anti-tumor composition. method.

26. A polynucleotide encoding the heavy or light chain, or an antigen-binding portion thereof, of any one of claims 1 to 16.

27. 27. A vector comprising the polynucleotide of claim 26, The vector is an expression vector selected from the group consisting of a mammalian expression vector, a yeast expression vector, an insect expression vector, and a bacterial expression vector. vector.

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