Methods of treating cancer using anti-C-C motif chemokine receptor 8 (CCR8) antibodies

Abstract

The present disclosure provides, inter alia, methods of treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject by administering an anti-CCR8 antibody to the subject in need thereof, and related therapeutic uses.

Description

[0001] Sequence Listing

[0002] This application contains a sequence listing, which has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The XML copy was created on October 5, 2023, named 50474-310WO3_Sequence_Listing_10_05_23, and is 161,635 bytes in size. Technical Field

[0003] The present invention relates to methods and compositions for use in treating cancer (eg, locally advanced, recurrent or metastatic solid tumor malignancies) in a patient. Background Art

[0004] Regulatory T (Treg) cells expressing the transcription factor Foxp3 are important for maintaining peripheral immune tolerance and preventing autoimmunity. Treg cells also constitute a major component of the immune infiltrate of solid cancers, promoting tumor development and progression by establishing an immunosuppressive tumor microenvironment and inhibiting anti-tumor immune responses. Treg cells can also hinder the efficacy of immunotherapy. An increased proportion of Treg cells in tumor-infiltrating lymphocytes has been associated with poor outcomes in several cancer indications.

[0005] Checkpoint blockade results in improved overall survival for some patients; however, there remains a subset of patients whose cancers are refractory to treatment (primary resistance) or progress after treatment (secondary resistance).

[0006] Therefore, there is a need for improved methods for treating cancer. Summary of the invention

[0007] The present disclosure provides, inter alia, methods of treating locally advanced, recurrent or metastatic solid tumor malignancies in subjects in need thereof, anti-CCR8 antibodies (e.g., monoclonal antibodies that bind to CCR8) for use in treating locally advanced, recurrent or metastatic solid tumor malignancies in subjects in need thereof, and related uses and kits.

[0008] In one aspect, the invention features a method of treating a locally advanced, recurrent, or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CC motif chemokine receptor 8 (CCR8).

[0009] In another aspect, the invention features a monoclonal antibody that binds to CCR8 for use in treating a locally advanced, recurrent, or metastatic solid tumor malignancy in a subject in need thereof.

[0010] In some aspects, (i) the subject has progressed after at least one available standard therapy; and / or (ii) the subject is a subject for whom all available standard therapies have proven ineffective or are intolerant or contraindicated.

[0011] In some aspects, locally advanced, recurrent or metastatic solid tumors are incurable.

[0012] In some aspects, the subject is 18 years of age or older.

[0013] In some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), melanoma, triple-negative breast cancer (TNBC), urothelial carcinoma (UC), esophageal cancer, gastric cancer, cervical cancer, renal cell carcinoma (RCC) or hepatocellular carcinoma (HCC).

[0014] In some aspects, the RCC is clear cell RCC.

[0015] In some aspects, the HNSCC is HNSCC of the oral cavity, oropharynx, hypopharynx, or larynx.

[0016] In some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is NSCLC.

[0017] In some aspects, the subject's tumor contains a targetable somatic alteration, and the subject has experienced disease progression during or after treatment with the targeted agent or is intolerant to the treatment.

[0018] In some aspects, targetable somatic alterations include somatic alterations involving epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), ROS proto-oncogene 1 (ROS1), proto-oncogene B-Raf (BRAF) V600E, neurotrophic tyrosine receptor kinase (NTRK), MET proto-oncogene (MET), RET proto-oncogene (RET), or Kirsten rat sarcoma virus (KRAS).

[0019] In some aspects, the melanoma is cutaneous melanoma.

[0020] In some aspects, the subject's tumor comprises a BRAFV600 mutation and the subject has experienced disease progression during or after treatment with or is intolerant to treatment with one or more serine / threonine protein kinase B-Raf (BRAF) inhibitors and / or one or more mitogen-activated protein kinase (MEK) inhibitors.

[0021] In some aspects, the locally advanced, recurrent, or metastatic solid tumor malignancy is UC.

[0022] In some aspects, the subject has: (i) histologically confirmed incurable advanced transitional cell carcinoma of the urothelium (including the renal pelvis, ureters, bladder, and urethra); and / or (ii) mixed histology, in which the subject's tumor has a dominant transitional cell pattern.

[0023] In some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is TNBC.

[0024] In some aspects, TNBC is defined by the American Society of Clinical Oncology-College of American Pathologists guidelines: (i) <1% of tumor cell nuclei are immunoreactive for estrogen receptor and <1% of tumor cell nuclei are immunoreactive for progesterone receptor; and / or (ii) HER2 negativity based on immunohistochemistry (IHC) and / or in situ hybridization.

[0025] In some aspects, the subject is checkpoint inhibitor (CPI) naive.

[0026] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is NSCLC or UC.

[0027] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is UC, wherein the subject is eligible for treatment with cisplatin, and the subject has experienced disease progression during or after treatment with cisplatin or is intolerant to the treatment.

[0028] In some aspects, the subject has not received prior treatment with a CPI, or where the subject has received adjuvant therapy with a CPI, the adjuvant therapy has been discontinued at least six months prior to the first administration of a monoclonal antibody that binds to CCR8 to the subject.

[0029] In some aspects, the subject has experienced a CPI.

[0030] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is NSCLC, HNSCC, melanoma, UC, TNBC, esophageal cancer, gastric cancer, cervical cancer, clear cell RCC, or HCC.

[0031] In some aspects, the subject receives clinical benefit from treatment comprising a PD-1 axis binding antagonist prior to disease progression.

[0032] In some aspects, the PD-1 axis binding antagonist is an anti-PD-L1 antibody or an anti-PD-1 antibody.

[0033] In some aspects, the subject has a treatment duration of greater than or equal to 6 months with a therapy comprising a PD-1 axis binding antagonist and / or has a partial response or a complete response as the best objective response.

[0034] In some aspects, (i) the subject has not been treated with a CPI, an immunomodulatory monoclonal antibody, or an immunomodulatory monoclonal antibody-derived therapy within 6 weeks prior to the first administration of a monoclonal antibody that binds to CCR8 to the subject; or (ii) the subject has previously been treated with a PD-1 axis binding antagonist, and the last administration of the PD-1 axis binding antagonist to the subject was at least 3 weeks prior to the first administration of the monoclonal antibody that binds to CCR8 to the subject.

[0035] In some aspects, the CPI is a PD-1 axis binding antagonist or a CTLA4 antagonist.

[0036] In some aspects, the PD-1 axis binding antagonist is an anti-PD-L1 antibody or an anti-PD-1 antibody.

[0037] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject in a dosing regimen that includes one or more dosing cycles.

[0038] In some aspects, the one or more dosing cycles include a 21 day dosing cycle.

[0039] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject on day 1 of each 21 -day dosing cycle.

[0040] In some aspects, a monoclonal antibody that binds to CCR8 is administered to a subject until disease progression or unacceptable toxicity occurs.

[0041] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject at a dose of 2 mg.

[0042] In some aspects, the monoclonal antibody that binds to CCR8 is administered intravenously to the subject.

[0043] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject intravenously by infusion.

[0044] In some aspects, a monoclonal antibody that binds to CCR8 is administered to a subject as a monotherapy.

[0045] In some aspects, a monoclonal antibody that binds to CCR8 is administered to a subject in combination with one or more additional therapeutic agents.

[0046] In some aspects, the one or more additional therapeutic agents comprises atezolizumab.

[0047] In some aspects, atezolizumab is administered to a subject in a dosing regimen comprising one or more dosing cycles.

[0048] In some aspects, the one or more dosing cycles include a 21 day dosing cycle.

[0049] In some aspects, atezolizumab is administered to the subject on day 1 of each 21 -day dosing cycle.

[0050] In some aspects, atezolizumab is administered to the subject at a dose of 1200 mg.

[0051] In some aspects, atezolizumab is administered intravenously to the subject.

[0052] In some aspects, atezolizumab is administered to the subject intravenously by infusion.

[0053] In some aspects, a tumor sample from a subject has been determined to have a detectable expression level of PD-L1.

[0054] In some aspects, a tumor sample from a subject has greater than or equal to 1% tumor cells (TC), immune cells (IC), comprehensive positive score (CPS), or tumor proportion score (TPS).

[0055] In some aspects, prior to administering atezolizumab to the subject, the subject has received at least two cycles of a monoclonal antibody that binds to CCR8.

[0056] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:29 or SEQ ID NO:30, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:31, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:32; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:26, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:27, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:28.

[0057] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0058] In some aspects, the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:106.

[0059] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 to 52; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0060] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35-47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48-52.

[0061] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:48; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0062] In some aspects, the monoclonal antibody that binds to CCR8 comprises: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:47; and (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:48.

[0063] In some aspects, VL comprises a V4M mutation, a P43A mutation, a F46L mutation, a C90Q mutation, or a combination thereof. In some embodiments, the V4M mutation, the P43A mutation, the F46L mutation, or the C90Q mutation is numbered according to Kabat.

[0064] In some aspects, VH comprises a G49S mutation, a K71R mutation, a S73N mutation, or a combination thereof. In some embodiments, the G49S mutation, the K71R mutation, or the S73N mutation is numbered according to Kabat.

[0065] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:55 and the light chain amino acid sequence of SEQ ID NO:56.

[0066] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:60 and the light chain amino acid sequence of SEQ ID NO:56.

[0067] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:111 and the light chain amino acid sequence of SEQ ID NO:56.

[0068] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:113 and the light chain amino acid sequence of SEQ ID NO:56.

[0069] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35-47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48-52.

[0070] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:47 and the VL sequence of SEQ ID NO:48.

[0071] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5, (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; and a light chain variable domain (VL) comprising (d) CDR-L1 comprising the amino acid sequence of SEQ ID NO:1, (e) CDR-L2 comprising the amino acid sequence of SEQ ID NO:2, and (f) CDR-L3 comprising the amino acid sequence of SEQ ID NO:3.

[0072] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0073] In some aspects, the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:106.

[0074] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0075] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 10-21 and a VL sequence selected from the group consisting of SEQ ID NOs: 22-25.

[0076] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:24; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0077] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:21; and a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:24.

[0078] In some aspects, the VL comprises a Y2I mutation. In some embodiments, the Y2I mutation is numbered according to Kabat.

[0079] In some aspects, VH comprises S73N mutation, V78L mutation, T76N mutation, F91Y mutation and P105Q mutation or its combination.In some embodiments, S73N mutation, V78L mutation, T76N mutation, F91Y mutation or P105Q mutation is according to Kabat numbering.

[0080] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:57 and the light chain amino acid sequence of SEQ ID NO:58.

[0081] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:61 and the light chain amino acid sequence of SEQ ID NO:58.

[0082] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:112 and the light chain amino acid sequence of SEQ ID NO:58.

[0083] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:114 and the light chain amino acid sequence of SEQ ID NO:58.

[0084] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:21 and the VL sequence of SEQ ID NO:24.

[0085] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 84, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 85; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 75.

[0086] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:95; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:94; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0087] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:95 and the VL sequence of SEQ ID NO:94.

[0088] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:101 and the light chain amino acid sequence of SEQ ID NO:100.

[0089] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:115 and a light chain amino acid sequence of SEQ ID NO:100.

[0090] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78.

[0091] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:97; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:96; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0092] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:97 and the VL sequence of SEQ ID NO:96.

[0093] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:103 and the light chain amino acid sequence of SEQ ID NO:102.

[0094] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:116 and a light chain amino acid sequence of SEQ ID NO:102.

[0095] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:90 or SEQ ID NO:91, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:92, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:93; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:79, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:80, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:81.

[0096] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:99; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:98; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0097] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:99 and the VL sequence of SEQ ID NO:98.

[0098] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:105 and a light chain amino acid sequence of SEQ ID NO:104.

[0099] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:117 and a light chain amino acid sequence of SEQ ID NO:104.

[0100] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0101] In some aspects, the antibody binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:106.

[0102] In some aspects, the antibody binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:106.

[0103] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:65 or SEQ ID NO:66, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:67, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:68; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:62, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:63, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:64.

[0104] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:70; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:69; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0105] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:70 and the VL sequence of SEQ ID NO:69.

[0106] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:72 and the light chain amino acid sequence of SEQ ID NO:71.

[0107] In some aspects, the monoclonal antibody that binds to CCR8 is a human antibody.

[0108] In some aspects, the monoclonal antibody that binds to CCR8 is a humanized antibody.

[0109] In some aspects, the monoclonal antibody that binds to CCR8 is a chimeric antibody.

[0110] In some aspects, the monoclonal antibody that binds to CCR8 is an antibody fragment that binds to CCR8.

[0111] In some aspects, the monoclonal antibody that binds to CCR8 is a full-length antibody.

[0112] In some aspects, the monoclonal antibody that binds to CCR8 is a full-length IgG1 antibody.

[0113] In some aspects, the monoclonal antibody that binds to CCR8 comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO:53 or SEQ ID NO:59.

[0114] In some aspects, the monoclonal antibody that binds to CCR8 comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO:54.

[0115] In some aspects, the monoclonal antibody that binds to CCR8 is present at about 1×10 -12 M is about 1×10 -11 The binding affinity (K d ) binds to CCR8.

[0116] In some aspects, the CCR8 is human CCR8.

[0117] In some aspects, the monoclonal antibody that binds to CCR8 is afucosylated.

[0118] In some aspects, the defucosylation ratio is between about 80% and about 95%.

[0119] In some aspects, regulatory T cells present in the tumor microenvironment of locally advanced, recurrent or metastatic solid tumor malignancies are depleted.

[0120] In some aspects, regulatory T cells outside the tumor microenvironment of locally advanced, recurrent or metastatic solid tumor malignancies are depleted.

[0121] In some aspects, the subject is a human. BRIEF DESCRIPTION OF THE DRAWINGS

[0122] Figure 1Screening results of anti-CCR8 monoclonal antibodies (mAbs) selectively binding to Treg cells (Treg) from human colorectal cancer dissociated tumor cells (DTCs) (obtained from Discovery LifeSciences) are depicted. Shown are CD8 T cells (defined as CD45+CD14-CD3+CD8+CD4-) (circles, ○), conventional CD4 T cells (defined as CD45+CD14-CD3+CD8-CD4+FOXP3-) (squares, □) and Treg cells (defined as CD45+CD14-CD3+CD8-CD4+FOXP3+) (triangles, ▲) mean fluorescence intensity (MFI) values. Three anti-CCR8 mAb clones in five anti-CCR8 mAb clones Ab1-Ab5 specifically stain intratumoral Treg cells, rather than conventional CD4 or CD8 T cells, and are sorted according to CCR8 MFI: hu.Ab4.H1L1>hu.Ab5.H1L1>hu.Ab3.H1L1.

[0123] Figure 2A and 2B Delineate the proposed mechanism of action of natural killer (NK) cell-mediated antibody-dependent cellular cytotoxicity (ADCC), which leads to the depletion of tumor-infiltrating CCR8-expressing Tregs ( Figure 2A ), and the ADCC activity of a human / cynomolgus monkey cross-reactive anti-CCR8 mAb proposed for further study ( Figure 2B EC of anti-CCR8 mAbs hu.Ab3.H1L1, hu.Ab5.H1L1, and hu.Ab4.H1L1 50 The values ​​were determined to be 0.02 nM, 0.02 nM and 0.08 nM, respectively.

[0124] Figures 3A to 3D Agonist and antagonist activities of human / cynomolgus monkey cross-reactive anti-CCR8 mAbs hu.Ab4.H1L1, hu.Ab5.H1L1 and hu.Ab3.H1L1 and comparator anti-CCR8 mAbs (humanized anti-human Yoshida anti-CCR8 antibody, mouse anti-human CCR8 mAb 433H (BD Biosciences) and mouse anti-human CCR8 mAb L263G8 (Biolegend) are depicted. Figure 3A As shown, CCL1, a known ligand for CCR8, exhibited agonist activity, but none of the anti-CCR8 tested mAbs showed antagonistic effects. Figure 3BThe data in show that anti-CCR8 mAb hu.Ab4.H1L1 exhibits antagonistic (neutralizing / ligand blocking) activity against the CCR8 ligand CCL1 (20 nM ligand), while anti-CCR8 mAbs hu.Ab5.H1L1 and hu.Ab3.H1L1 do not exhibit ligand blocking (non-neutralizing) activity at the concentrations studied. Figure 3C The data in show that comparator anti-CCR8 mAbs (humanized anti-human Yoshida anti-CCR8 antibody, mouse anti-human CCR8 mAb 433H (BD Biosciences), and mouse anti-human CCR8 mAb L263G8 (Biolegend)) did not show agonistic effects, whereas the CCR8 ligand CCL1 showed agonistic effects. Figure 3D The data in show that the comparator anti-CCR8 mAbs (humanized anti-human Yoshida anti-CCR8 antibody, mouse anti-human CCR8 mAb 433H (BD Biosciences) and mouse anti-human Biolegend L263G8 (Biolegend)) exhibit antagonistic (neutralizing / ligand blocking) activity against the CCR8 ligand CCL1. The IC values ​​for ligand blocking activity are provided in the Examples. 50 value.

[0125] Figures 4A to 4F Depicts hu.Ab3.H1L1( Figure 4A )、hu.Ab4.H1L1( Figure 4B ) and hu.Ab5.H1L1( Figure 4C ) and commercial anti-CCR8 mAb mouse anti-human CCR8 mAb 433H (BD Biosciences) ( Figure 4D ) and mouse anti-human CCR8 mAb L263G8 (Biolegend) ( Figure 4E ) and humanized anti-human Yoshida anti-CCR8 mAb ( Figure 4F ) and the N-terminal end of the human GPCRs (CCR2, CCR3, CCR4, CCR5, CCR8, CXCR4, ACKR2 and ACKR4) (DYKDDDDK, SEQ ID NO: 118)-tagged plasmids, hCCR8 constructs or using Binding data of HEK293 cells transiently transfected with the mock construct (reagent:DNA=3:1). Antibody control (5ug / mL) staining was used to confirm cell surface expression of each GPCR. mAbs hu.Ab4.H1L1 and hu.Ab5.H1L1 stained only hCCR8-containing cells, confirming their specificity for hCCR8. mAb hu.Ab3.H1L1 showed staining of multiple other GPCRs, indicating a lack of specificity. The CCR8-selective hu.Ab4.H1L1 and hu.Ab5.H1L1 mAbs (e.g., Figure 2A and 2B continued to be used for further research.

[0126] Figures 5A to 5D The light chain variable regions of the sequences of the rabbit (rb.Ab4) and humanized Ab4 (L1-L4 and H1-H12) CCR8 mAbs studied are depicted ( Figure 5A ) and the heavy chain variable region ( Figures 5B to 5D ) comparison. Based on the binding evaluation of the variant antibodies, Y2 on the light chain (L3) and S73, T76, V78, F91 and P105 on the heavy chain (H12) were determined to be key rabbit vernier residues. CDRs, variable regions, constant regions and full-length sequences are provided in the examples.

[0127] Figures 6A to 6D The light chain variable regions of the sequences of the rabbit (rb.Ab5) and humanized Ab5 (L1-L5 and H1-H13) CCR8 mAbs studied are depicted ( Fig. 6A ) and the heavy chain variable region ( Figures 6B to 6D ) comparison. The C90Q mutation in CDR L3 was introduced to remove an unpaired cysteine ​​that would be a disadvantage during manufacturing. Based on the binding evaluation of the variant antibodies, V4, P43 and F46 on the light chain (L1) and G49, K71 and S73 (H13) on the heavy chain were determined to be key rabbit vernier residues. CDRs, variable regions, constant regions and full-length sequences are provided in the examples.

[0128] Figures 7A to 7D Depicted are the results of cell-based affinity measurements of hu.Ab5.H13L1 and hu.Ab4.H12L3 mAbs using radiolabeled IgG and CHO cell lines stably expressing either human CCR8 or cynomolgus ("cyno") CCR8. The data demonstrate that hu.Ab4.H12L3 and hu.Ab5.H13L1 mAbs have similar affinities for human and cynomolgus CCR8, indicating the expected cross-reactivity (compare Fig. 7A and Figure 7B , and compare Figure 7C and Fig.7D ). Kd (nM) affinity data from these studies are provided in the Examples.

[0129] Fig. 8A and 8B Depicts hu.Ab4.H12L3( Fig. 8A ) and hu.Ab5.H13L1( Figure 8B ) mAb binding data to a panel of sulfated GPCRs and again confirmed that these Ab4 and Ab5 variants (with Figure 4B and Figure 4C Ab4 and Ab5 data provided in the literature) showed selectivity for CCR8. The tag binds weakly (it affects the binding of the N-terminal epitope of Ab5; see Fig.16A and 16B ), thus having a C-terminal The CCR8 constructs are also provided at Figure 4C middle.

[0130] Fig. 9A and 9B Effects of anti-CCR8 mAbs hu.Ab4.H12L3 and hu.Ab5.H13L1 on CCR8 activation as measured by Ca 2+ Influx assay ( Fig. 9A ) and CCR8 CCL1 ligand binding ( Fig. 9B ) is determined by Figure 3A The data is similar, Fig. 9A Again, it was confirmed that neither Ab4 nor Ab5 anti-CCR8 mAb variants showed agonism in the absence of the CCR8 ligand CCL1. Figure 3B The data is similar, Fig. 9B It was again demonstrated that the Ab4 variant exhibited antagonism against the CCR8 ligand CCL1 (20 nM ligand), whereas the Ab5 variant exhibited no ligand blocking activity at the concentrations studied. The IC values ​​for ligand blocking activity are provided in the Examples. 50 value.

[0131] Figures 10A to 10E Depicted are the staining differences of hu.Ab4.H12L3 and hu.Ab5.H13L1 on CCR8+ HEK293 cells with (hCCR8.TPST1 / 2NTC) and without tyrosyl-protein sulfotransferase (TPST) 1 and tyrosyl-protein sulfotransferase (TPST) 2 (hCCR8.TPST1 / 2KO) compared to humanized anti-human Yoshida CCR8 mAb and commercial antibodies mouse anti-human CCR8 mAb433H (BD Biosciences) and mouse anti-human CCR8 mAb L263G8 (Biolegend). Fig. 10A) and hu.Ab5.H13L1( Fig. 10B ) showed similar binding / staining to both cell lines (hCCR8.TPST1 / 2NTC and hCCR8.TPST1 / 2KO), indicating that they bind CCR8 independently of tyrosine sulfation (“sulfation-independent”). In contrast, the humanized anti-human Yoshida CCR8 antibody ( Fig. 10C ) and commercial antibody mouse anti-human CCR8 mAb 433H (BD Biosciences) ( Fig. 10D ) and mouse anti-human CCR8 mAb L263G8 (Biolegend) ( Fig. 10E ) failed to bind to TPST1 / 2KO cells, indicating that they require tyrosine sulfation of CCR8 for binding and are therefore considered “sulfation-dependent”.

[0132] Figures 11A to 11D Depicted is the effect of defucosylated CCR8 mAbs Afuc.hu.Ab5.H13L1 and Afuc.hu.Ab4.H12L3 on CHO cells stably expressing hCCR8 (using NK-92F158 ( Fig.11A ) and NK-92V158( Fig. 11B ) as effector cells) showed enhanced (>10-fold improvement) ADCC activity and compared with humanized anti-human Yoshida anti-CCR8 antibody ( Fig. 11C ) also showed 10- to 20-fold improved ADCC activity compared to the commercial anti-CCR8 mAbs mouse anti-human CCR8 mAb 433H (BD Biosciences) and mouse anti-human CCR8 mAb L263G8 (Biolegend) that were demonstrated to have no ADCC activity (as expected) since the assay used is primarily relevant to antibodies containing human Fc regions ( Fig. 11C ). Fig.11D Murine anti-human CCR8 mAb 433H (BD Biosciences) and Murine anti-human CCR8 mAb L263G8 (Biolegend) were shown to have ADCC activity and human anti-CCR8 activity using an assay specific for antibodies containing a murine Fc region. Activity data are also provided in the Examples.

[0133] Figures 12A to 12DFigure 3 shows the selective ADCC activity against human Treg cells compared to conventional human CD4 T cells from peripheral blood mononuclear cells (PBMCs) transferred to NOD.Cg-Prkdc when incubated with afucosylated, fucosylated (hIgG1) and afucosylated isotype control mAb ("gD.afuc") and primary NK cells as effector cells. scid Iq tm1Wjl / SzJ(NSG TM ) mice were induced to express CCR8 and then recovered. ADCC activity against Treg cells was measured by calculating the ratio of recovered Treg cells to recovered CD8 cells (Treg / CD8) or conventional CD4 T cells to recovered CD8 T cells (CD4conv / CD8). Fig. 12B ), CCR8 mAbs Afuc.hu.Ab4.H12L3 and hu.Ab4.H12L3 selectively mediated ADCC activity against Treg cells ( Fig. 12A ), where the defucosylated variant exhibited increased ADCC activity. Similarly, compared with conventional CD4 T cells ( Fig.12D ), CCR8 mAb Afuc.hu.Ab5.H13L1 and hu.Ab5.H13L1 selectively mediated ADCC activity against Treg cells ( Fig. 12C ), wherein the defucosylated variant exhibited increased ADCC activity.

[0134] Figures 13A to 13D Depicted are selective ADCC activity against Treg cells compared to conventional CD4 T cells when human dissociated renal cell carcinoma (RCC) cells were incubated with afucosylated, fucosylated (hIgG1), and afucosylated isotype control mAb ("gD.afuc") and primary NK cells as effector cells. ADCC activity against Treg cells was measured by calculating the ratio of recovered Treg cells to recovered CD8 cells (Treg / CD8) or conventional CD4 T cells to recovered CD8 T cells (CD4conv / CD8). ADCC activity against Treg cells was measured by calculating the ratio of recovered Treg cells to recovered CD8 T cells (Treg / CD8) or conventional CD4 T cells to recovered CD8 T cells (CD4conv / CD8). Fig. 13B ), CCR8 mAbAfuc.hu.Ab4.H12L3 and hu.Ab4.H12L3 selectively mediated ADCC activity against Treg cells ( Fig.13A ), where the defucosylated variant exhibited increased ADCC activity. Similarly, compared with conventional CD4 T cells ( Fig.13D), CCR8 mAbAfuc.hu.Ab5.H13L1 and hu.Ab5.H13L1 selectively mediated ADCC activity against Treg cells ( Fig. 13C ), wherein the defucosylated variant exhibited increased ADCC activity.

[0135] Figures 14A to 14E It was shown that the defucosylated anti-CCR8 mAbs Afuc.hu.Ab5.H13L1 and Afuc.hu.Ab4.H12L3 were significantly more effective in the presence of HR / FF ( Fig.14A )、RR / FF( Fig. 14B )、HR / VF( Fig. 14C ) and RR / VF( Fig.14D CD14 of four different donors with the FcgRIIa(H131R) / FcgRIIIa(V158F) genotype + It exhibits enhanced ADCP activity in monocyte-derived macrophages and is comparable to humanized anti-human Yoshida anti-CCR8 antibody ( Fig.14E ) also showed 3 to 4 times improved ADCP activity. Activity data are also provided in the examples.

[0136] Figures 15A to 15D Compared with the FcgRIIa enhanced G236A.I332E variant Afuc.hu.Ab5.H13L1.G236A.I332E, the defucosylated anti-CCR8 mAb Afuc.hu.Ab5.H13L1 was significantly more effective in the presence of HR / FF ( Fig.15A )、RR / FF( Fig. 15B )、HR / VF( Fig. 15C ) and RR / VF( Fig.15D CD14 of four different donors with the FcgRIIa(H131R) / FcgRIIIa(V158F) genotype + Monocyte-derived macrophages exhibit enhanced ADCP activity.

[0137] Fig.16A and 16B Depicts hu.Ab5.H13L1( Fig.16A ) and hu.Ab4.H12L3( Fig. 16B ) epitope map of mAb. Fig.16A As shown, a construct was generated encoding a C-terminal hu.Ab5.H13L1 did not bind to D2A, Y3A, L5A, and D6A when individual alanine point mutations were present at positions 2-24 in the tagged hCCR8, indicating that the epitope includes at least the DYTLD region at the N-terminus of human CCR8. Fig. 16B As shown, a construct was generated that encodes a human CCR8.CCR5 chimera (N-term1, N-term2, ECL1, ECL2, and ECL3) in which different extracellular regions of hCCR8 were replaced by a C-terminal The corresponding region of CCR5 of the tag was replaced, and hu.Ab4.H12L3 did not bind to the ECL1 and ECL2 chimera, indicating that the epitope of this antibody includes at least the ECL1 and ECL2 regions of the CCR8.huCCR8 N-terminus: MDYTLDLSVTTVTDYYYPDIFSSP (SEQ ID NO: 110).

[0138] Figures 17A to 17I Figure 3 shows tumor growth in CT26 tumor-bearing mice three days after injection of a single dose of mouse surrogate anti-CCR8 mAb at increasing concentrations between 0.003 mg / kg and 5 mg / kg. Fig.17A ) but not in the spleen ( Fig. 17B ) or tumor-draining lymph nodes ( Fig. 17C ) did not result in progressive depletion of Treg cells. Anti-CCR8 mAb treatment did not lead to CD4 conventional T cells ( Figures 17D to 17F ) or CD8 T cell depletion ( Figures 17G to 17I ). An isotype control antibody (anti-gp120) was used.

[0139] Fig.17J Depicts depletion of Treg cells in tumors (measured as the fraction of Treg cells with CD45+ leukocytes) but not in spleen, draining lymph nodes (dLN), or blood three days after injection of mice bearing E0771 syngeneic tumors with a single dose of anti-CCR8 antibody at increasing concentrations between 0.01 g / kg and 1 mg / kg.

[0140] Figures 17K to 17O A minimal physiologically based pharmacokinetic-pharmacodynamic (PBPK-PD) model for simulating the pharmacokinetic (PK) / receptor occupancy (RO) relationship as well as the pharmacodynamics (PD) and efficacy of a surrogate anti-CCR8 antibody in mice is depicted. The minimal PBPK model structure is shown in Figure 17K As shown. This model predicts the PK of anti-CCR8 antibody ( Figure 17L )、RO( Fig.17M ), Treg depletion ( Fig.17N ) and antitumor efficacy ( Fig.17O ).

[0141] Figures 18A to 18D Depicted are the results of the comparison with anti-CD25 mAb ( Fig.18D ) or isotype control mAb (anti-gp120) ( Fig.18A ) in mice with established CT26 syngeneic tumors, compared with a single dose of ( Fig.18B ) or twice weekly ( Fig.18C ) mice treated with alternative anti-CCR8 mAb for tumor growth inhibition. When the tumor volume reached 150-250mm 3 Treatment was started at 9:00 p.m. Tumor volumes were measured over time. Gray lines represent individual mice and black lines represent group fits.

[0142] Figures 19A to 19E Depicts the time of tumor inoculation ( Fig.19B ) or the tumor reaches 150 to 250 mm 3 hour( Fig.19D ) administration of effector-competent mice to replace the growth inhibition of CT26 tumors observed with anti-CCR8 mAb. In the same ligand-blocking anti-CCR8 mAb, the effector-incompetent LALAPG variant ( Fig.19C and 19E ). Tumor volumes were measured over time. Gray lines represent individual mice and black lines represent group fits. Isotype control mAb (anti-gp120) ( Fig.19A ).

[0143] Figures 20A to 20D showed that the combination of anti-CCR8 mAb and anti-PDL1 mAb in mice ( Fig.20D ) unexpectedly showed superior growth inhibition of EMT6 tumors compared to anti-CCR8 mAb alone ( Fig. 20B ) or anti-PDL1 mAb alone ( Fig. 20C ) is more effective. When the tumor reaches 150-250mm 3 Treatment was started at 9:00 pm. Tumor volumes were measured over time. Gray lines represent individual mice and black lines represent group fits. Isotype control mAb (anti-gp120) ( Fig. 20A ).

[0144] Fig.21Depicted are serum pharmacokinetic profiles (mean ± SD) of anti-gD (control) and test anti-CCR8 mAbs Afuc.hu.Ab5.H13L1 and Afuc.hu.Ab4.H12L3 in cynomolgus monkeys after a single dose of 10 mg / kg IV bolus. Afuc.hu.Ab5.H13L1 exhibited desirable sustained serum concentration levels over a 35-day period post-dose, which is expected to elicit more sustained target engagement, which may translate into better anticancer activity and less frequent dosing.

[0145] Figures 22A to 22C Depicted are the responses to 10 mg / kg defucosylated anti-gD injected intravenously (control; group 1, designated 1001, 1002, 1003; Fig.22A ), Afuc.hu.Ab5.H13L1 (Group 2, designated 2001, 2002, 2003; Fig. 22B ) or Afuc.hu.Ab4.H12L3 (Group 3, designated 3001, 3002, 3003; Fig. 22C Figure 2. Whole blood flow cytometric analysis of total Treg cell counts in nine male cynomolgus monkeys treated with 100 mg / kg CCR8 mAbs. Neither of the two tested anti-CCR8 mAbs significantly reduced the absolute counts of total T-reg cells in whole blood up to 840 hours after dosing.

[0146] Figures 23A to 23I Depicted are the results of the experiments against defucosylated anti-gD (control; group 1, designated as 1001 ( Fig.23A )、1002( Fig. 23B )、1003( Fig.23C )), Afuc.hu.Ab4.H12L3 (Group 3, designated 3001 ( Fig.23D )、3002( Fig.23E )、3003( Fig.23F )) or Afuc.hu.Ab5.H13L1 (Group 2, designated 2001 ( Figure 23G )、2002( Fig.23H )、2003( Fig.23I)) Results of whole blood flow cytometric analysis of CCR8+FoxP3+Treg cell reduction in 9 male cynomolgus monkeys dosed with . Blood was collected from each animal before dosing ("pre-study") and at 0 hours on day 1 ("pre-dose"). Each animal was then administered a single dose of 10 mg / kg defucosylated anti-gD (control group), Afuc.hu.Ab5.H13L1 (Group 2) or Afuc.hu.Ab4.H12L3 (Group 3) via intravenous injection. Blood was then collected from the animals and subjected to the following treatments prior to flow cytometric analysis: (i) blood samples not spiked with any of the tested CCR8 mAbs ("unspiked"), (ii) blood samples further spiked with saturating concentrations of Afuc.hu.Ab5.H13L1, and (iii) blood samples further spiked with saturating concentrations of Afuc.hu.Ab4.H12L3. Each sample in the unspiked and spiked samples was subsequently treated with labeled goat anti-human IgG antibody and analyzed by flow cytometry. Figures 23A to 23C As can be seen in the Figure 1, the control (Group 1) was initially treated, but flow cytometry of unspiked blood demonstrated that total CCR8+ T-reg cells were not regulated. In addition, flow cytometry of spiked blood had little effect on total CCR8+ T-reg cell counts. Regarding Group 3, as can be seen from Figures 23D to 23F As can be seen in Figure 2, flow cytometric analysis of the blood of each of the three animals showed a decrease in CCR8+ T-reg cells up to 168 hours after dosing. Figures 23G to 23I As can be seen in Figure 2, flow cytometry analysis of blood showed a decrease in CCR8+ T-reg cells in animals 2002 and 2003. Group 2 and 3 animals showed little effect on overall Treg cell counts ( Figures 22A to 22C ), but showed a reduced number of peripheral blood CCR8+T-reg cells after administration ( Figures 23D to 23I ) (both in the spiked and unspiked cases), which is consistent with the proposed mechanism of action (see Figure 2A ) are consistent.

[0147] Fig.24 Schematic diagram showing the study design for the Phase Ia portion of the GO43860 study with RO7502175 as a single agent. DL = dose level; DLT = dose-limiting toxicity; HCC = hepatocellular carcinoma; HNSCC = head and neck squamous cell carcinoma; MAD = maximum administered dose; MTD = maximum tolerated dose; NSCLC = non-small cell lung cancer; PD = pharmacodynamics; PK = pharmacokinetics; Q3W = every 3 weeks; RCC = renal cell carcinoma; TNBC = triple-negative breast cancer; UC = urothelial carcinoma. Actual number of patients may vary.

[0148] Fig.25Schematic diagram showing the study design for the Phase Ib portion of the GO43860 study (RO7502175 in combination with atezolizumab). TBD = To Be Determined.

[0149] Fig.26 Schematic diagram showing the conditions for continued treatment after disease progression in the GO43860 study. ECOG = Eastern Cooperative Oncology Group; RECIST = Response Evaluation Criteria in Solid Tumors.

[0150] Fig.27A and 27B Schematic diagram showing the conditions for patients in phase Ia to cross over from phase Ia to phase Ib in the GO43860 study. AE = adverse event; PD = progressive disease.

[0151] Figures 28A to 28C The in vitro pharmacology of RO7502175 is described. Fig.28A )RO7502175 only bound to CCR8 from humans and cynomolgus monkeys among the species assessed. Defucosylated RO7502175 showed binding to FcγRIIIA-F158 ( Fig.28B ) and FcγRIIIA-V158( Fig.28C )'s enhanced combination.

[0152] Fig.29 Depicted is the in vitro ADCC activity of RO7502175 on human Treg cells from pre-activated PBMCs. Shown is the ADCC activity of RO7502175, fucosylated (wild type) anti-CCR8 control and defucosylated anti-gD (isotype) control on human Treg cells isolated from pre-activated PBMCs from three different donors.

[0153] Fig. 30A and 30B The in vitro ADCC activity of RO7502175 against dissociated tumor cells and CHO cells expressing CCR8 is depicted. Fig. 30A ) The figure shows the ADCC activity of RO7502175 on human regulatory, conventional CD4+ and CD8+ T cells from dissociated renal cell carcinoma tumors. ( Fig. 30B ) The figure shows the percentage of cytotoxicity of RO7502175 in the ADCC assay using CHO cells expressing CCR8 (using PBMCs isolated from 8 different healthy donors) (mean ± SD). The geometric mean EC50 values ​​are listed.

[0154] Figures 31A to 31F Depicted is the in vitro cytokine release of RO7502175 in a PBMC assay using soluble and immobilized forms. Fig.31A ) soluble assay form and ( Fig.31B ) Graphical display of the immobilized assay format combining test article and PBMC. Figures 31C to 31F ) shows the interaction of the indicated test articles in soluble or immobilized form with PBMC (n=8), ( Fig.31C )IFNγ、( Fig.31D )IL-2, ( Fig.31E )IL-6 and ( Fig.31F ) Cytokine concentration (pg / mL) after 18 h of TNFα incubation.

[0155] Figures 32A to 32E Depicted is the in vivo activity of anti-murine CCR8 antibodies in a syngeneic mouse tumor model. Fig.32A ) Study design: C57BL / 6 mice bearing E0771 tumors were given a single IV administration of anti-mouse CCR8 antibody and ( Fig.32B )Treg cells and ( Fig.32C )CD8+T cell frequency and ( Fig.32D ) Serum antibody concentration (mean ± SD). Fig.32E ) shows the antitumor activity of E0771 tumor-bearing C57BL / 6 mice after a single IV administration of anti-murine CCR8 antibody in the efficacy study. dLN, draining lymph node; MQC, minimum quantifiable concentration.

[0156] Fig.33A and 33B Describe the in vivo activity of anti-murine CCR8 antibodies in a syngeneic mouse tumor model (from Figures 32A to 32D Additional data from the PD study described in ). Fig.33A )Treg cells and ( Fig.33B )The frequency of CD8+ T cells in the spleen and draining lymph nodes.

[0157] Fig.34 Depicted from Fig.32E PK of anti-murine CCR8 antibodies in C57BL / 6 mice from the efficacy study described in . Serum concentrations (mean ± SD) estimated from pharmacokinetic parameters of non-compartmental analysis after a single IV administration of anti-murine CCR8 antibodies in C57BL / 6 mice are shown.

[0158] Figures 35A to 35D The PK, PD, and safety profiles of RO7502175 in cynomolgus monkeys were characterized. Fig.35A ) shows the serum concentration-time curves (mean ± SD) and ( Fig.35B ) Plasma MCP-1 and IL-6 cytokine concentrations (mean ± SD). Fig.35C) shows the serum concentration-time curves (mean ± SD) in cynomolgus monkeys following IV administration of vehicle control or RO7502175 in a repeated dose study and ( Fig.35D ) Fold change of CCR8+ Treg cells in blood compared with baseline (mean ± SD).

[0159] Fig.36A and 36B Depicted is the PD curve of RO7502175 in a single-dose study in cynomolgus monkeys. Fig.36A ) anti-gD (control) or ( Fig.36B ) Relative percentage of CCR8+ Treg cells in the blood of individual male cynomolgus monkeys after RO7502175 (mean±SEM; n=2 replicates).

[0160] Figures 37A to 37H The mPBPK-PD model reproduced preclinical PK, PD and anti-tumor efficacy data. Fig.37A ) shows a schematic diagram of the mPBPK-PD model structure. ( Fig.37B ) The model captured the anti-murine CCR8 antibody PK in mice following single-dose administration at 0.01, 0.03, 0.1, and 1 mg / kg IV. ( Fig.37C ) model predicts receptor occupancy (RO) of anti-mouse CCR8 antibodies over time. Fig.37D ) The model was calibrated based on tumor CCR8+Treg cell counts. Fig.37E ) model captures the increase in CD8+ T cells and ( Fig.37F )Average tumor killing. ( Figure 37G ) The model was calibrated and validated based on the PK of RO7502175 in single-dose and repeated-dose studies in cynomolgus monkeys. Fig.37H ) shows RO predictions for cynomolgus monkeys at 10 mg / kg single dose and 30 and 100 mg / kg qw dosing. Symbols represent individual data points, and lines represent model fits or predictions.

[0161] Figures 38A to 38D The predicted RO7502175 clinical PK profile and receptor occupancy (RO) in patient plasma and tumors are depicted. Blood ( Fig.38A and 38B ) and tumor compartments ( Fig.38C and 38D ) in clinical PK and RO prediction.

[0162] Fig.39Depicted is the expected RO7502175 receptor occupancy (RO) in the patient's tumor. Shown is the clinical mean RO prediction for the tumor compartment in cycle 1 (21 days) after administration of RO7502175 at dose levels of 0.2, 0.6, 2, 6, and 20 mg IV q3w. These ranges show the mean RO of the tumor taking into account the uncertainty of the PK parameters (V plasma = 37.1 mL / kg + / - 20%, CL = 0.12 mL / h / kg + / - 20%, sig_tumor = 0.91 to 0.95 (for 5% to 10% tumor / plasma AUC), K D =0.032 nM to 0.060 nM). Vplasma, plasma volume; CL, clearance; sig_tumor, tumor reflectivity; K D , antibody binding affinity to CCR8.

[0163] Fig.40A and 40B Depicted is the first-in-human dose selection for RO7502175. Fig.40A ) A MABEL-based approach, an mPAD-based approach, and a comprehensive approach based on all preclinical data were considered in selecting the RO7502175 FiH dose. The workflow in the schematic shows the in vitro and in vivo datasets used as input and the criteria for the results generated according to each method evaluated. ( Fig.40B ) The figure shows the FiH dose of RO7502175 based on each method examined. RO, receptor occupancy; K D , binding affinity of antibody to CCR8; ADCC, antibody-dependent cell-mediated cytotoxicity; MABEL, minimum expected biological effect level; C max , maximum observed concentration; mPAD, minimum pharmacologically active dose; C avg , mean concentration over a defined time period after administration; NOAEL, no observed adverse effect level; CRA, in vitro cytokine release assay. DETAILED DESCRIPTION

[0164] I. Definitions

[0165] For purposes herein, "acceptor human framework" is such a framework, which comprises the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. The acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework can comprise the amino acid sequence identical to the human immunoglobulin framework or a human consensus framework, or it can contain amino acid sequence changes. In some aspects, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less or 2 or less. In some aspects, the VL acceptor human framework is identical to the VL human immunoglobulin framework sequence or a human consensus framework sequence in sequence.

[0166] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (K D ) represents. Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary methods for measuring binding affinity will be described herein.

[0167] An "affinity matured" antibody is one with one or more alterations in one or more complementarity determining regions (CDRs) which result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess such alterations.

[0168] The terms "anti-CCR8 antibody" and "antibody that binds to CCR8" refer to an antibody that is capable of binding to CCR8 with sufficient affinity so that the antibody can be used as a diagnostic and / or therapeutic agent targeting CCR8. In one aspect, the extent of binding of an anti-CCR8 antibody to unrelated, non-CCR8 proteins is less than about 10% of the binding of the antibody to CCR8, as measured, for example, by surface plasmon resonance (SPR). In certain aspects, the dissociation constant (K) of an antibody that binds to CCR8 is D ) is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM or ≤0.001 nM (e.g., 10 -8 M or smaller, e.g. 10 -13 M to 10 -8 M, for example 10 -13 M to 10 -9 M). In some aspects, the antibody that binds to CCR8 has a specificity of about 1×10 -12M is about 1×10 -10 M, about 1×10 -12 M is about 1×10 -11 M, or about 1×10 -11 M is about 5×10 -11 M of K D In some aspects, the antibody that binds to CCR8 has a CCR8 affinity of about 2×10 -11 M of K D In some aspects, the antibody that binds to CCR8 has a CCR8 affinity of about 5×10 -12 M of K D When the antibody has a K of 1 μM or less D When the antibody "specifically binds" to CCR8, the antibody is said to "specifically bind" to CCR8. In certain embodiments, the anti-CCR8 antibody binds to an epitope of CCR8 in at least two different species (e.g., human and cynomolgus monkey).

[0169] The term "antibody" herein is used in the broadest sense and includes various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0170] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody and binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') 2 ; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv and scFab); single domain antibodies (dAb); and multispecific antibodies formed from antibody fragments. For a review of certain antibody fragments, see Holliger and Hudson, Nature Biotechnology (2005) 23: 1126-1136.

[0171] The term "epitope" refers to a site on a protein or non-protein antigen to which an anti-CCR8 antibody binds. An epitope may be formed by a continuous stretch of amino acids (linear epitope) or by the inclusion of non-continuous amino acids (conformational epitope), for example due to antigen folding, i.e., due to the tertiary folding of the protein antigen, which is in close spatial proximity. After the protein antigen is exposed to a denaturant, a linear epitope is usually still bound by an anti-CCR8 antibody, while a conformational epitope is usually destroyed after treatment with a denaturant. An epitope comprises at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 10, at least 15, at least 20, at least 30, or at least 35 or 3 to 25, 3 to 20, 3 to 15, 3 to 10, 3 to 5, 30 to 40, 35 to 40, or 5 to 10 amino acids in a unique spatial configuration.

[0172] Screening for antibodies that bind to a specific epitope (i.e., those that bind to the same epitope) can be performed using methods routine in the art, such as, for example, but not limited to, alanine scanning, peptide blotting (see Kobeissy et al., Meth. Mol. Biol. (2004) 248:443-463), peptide cleavage analysis, epitope excision, epitope extraction, chemical modification of antigens (see Hochleitner et al., Prot. Sci. 9 (2000) 487-496), and cross-blocking (see "Antibodies", Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harb., NY)).

[0173] Antigen structure-based antibody profiling (ASAP), also known as modification-assisted profiling (MAP), allows binning of a large number of monoclonal antibodies that specifically bind to CCR8 based on the binding profile of each antibody to a chemically or enzymatically modified antigen surface (see, e.g., US2004 / 0101920). The antibodies in each group bind to the same epitope, which may be a unique epitope that is significantly different from or partially overlaps with the epitope represented by another group.

[0174] Competitive binding can also be used to easily determine whether an antibody binds to the same epitope of CCR8 or competes for binding with a reference anti-CCR8 antibody. For example, an "antibody that binds to the same epitope" as a reference anti-CCR8 antibody refers to an antibody that blocks the binding of the reference anti-CCR8 antibody to its antigen by 50% or more in a competition assay, whereas the reference antibody blocks the binding of the antibody to its antigen by 50% or more in a competition assay. Also for example, to determine whether an antibody binds to the same epitope as a reference anti-CCR8 antibody, the reference antibody is allowed to bind to CCR8 under saturating conditions. After removing excess reference anti-CCR8 antibody, the ability of the anti-CCR8 antibody in question to bind to CCR8 is assessed. If the anti-CCR8 antibody is able to bind to CCR8 after saturation binding of the reference anti-CCR8 antibody, it can be concluded that the anti-CCR8 antibody in question binds to a different epitope than the reference anti-CCR8 antibody. However, if the anti-CCR8 antibody in question is unable to bind to CCR8 after saturation binding of the reference anti-CCR8 antibody, the anti-CCR8 antibody in question may bind to the same epitope as the epitope bound by the reference anti-CCR8 antibody. In order to confirm whether the antibody in question binds to the same epitope or is blocked for steric reasons, routine experiments (e.g., peptide mutations and binding analysis using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art) can be used. The assay should be performed in two settings, i.e., both antibodies are saturated antibodies. If in both settings, only the first (saturated) antibody is able to bind to CCR8, it can be concluded that the anti-CCR8 antibody in question and the reference anti-CCR8 antibody compete for binding to CCR8.

[0175] In some aspects, two antibodies are considered to bind to the same or overlapping epitope if a 1, 5, 10, 20, or 100-fold excess of one antibody inhibits the binding of the other antibody by at least 50%, at least 75%, at least 90%, or even 99% or more as measured in a competitive binding assay (see, e.g., Junghans et al., Cancer Res. 50 (1990) 1495-1502).

[0176] In some aspects, two antibodies are considered to bind to the same epitope if substantially all amino acid mutations in the antigen that reduce or eliminate binding of one antibody also reduce or eliminate binding of the other antibody. Two antibodies are considered to have "overlapping epitopes" if only a subset of amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody.

[0177] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0178] The "class" of an antibody refers to the type of constant domain or region possessed by the heavy chain of the antibody. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of them can be further divided into subclasses (isotypes), e.g., IgG 1 IgG 2 IgG 3 IgG 4 , IgA 1 , and IgA 2 In certain aspects, the antibody is IgG 1 Isotype. In certain aspects, the antibody is an IgG with P329G, L234A and L235A mutations to reduce the effector function of the Fc region. 1 Isotype. In other aspects, the antibody is IgG 2 Isotype. In certain aspects, the antibody is an IgG with an S228P mutation in the hinge region. 4 Isotype to improve IgG 4 Stability of antibodies. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ. The light chains of antibodies can be classified into one of two types called kappa (κ) and lambda (λ) based on the amino acid sequence of their constant domains.

[0179] As used in the present application, the term "human-derived constant region" or "human constant region" refers to the constant heavy chain region and / or constant light chain kappa or lambda region of a human antibody of subclass IgG1, IgG2, IgG3 or IgG4. Such constant regions are well known in the prior art and are described, for example, by Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991) (see also, for example, Johnson, G. and Wu, TT, Nucleic Acids Res. 28 (2000) 214-218; Kabat, EA et al., Proc. Natl. Acad. Sci. USA 72 (1975) 2785-2788). Unless otherwise indicated herein, the numbering of the amino acid regions within the constant region is according to the EU numbering system, also known as the EU index of Kabat, as described in Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991), NIH Publication No. 91-3242.

[0180] "Effector functions" refer to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[0181] An "effective amount" of an agent (eg, a pharmaceutical composition) refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

[0182] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, which comprises at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one aspect, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the antibody produced by the host cell can undergo post-translational cleavage of one or more (particularly one or two) amino acids from the C-terminus of the heavy chain. Therefore, the antibody produced by the host cell by expressing a specific nucleic acid molecule encoding a full-length heavy chain can include a full-length heavy chain, or the antibody can include a cleavage variant of the full-length heavy chain. This may be a situation where the last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, EU numbering system). Therefore, the C-terminal lysine (Lys447) or C-terminal glycine (Gly446) and lysine (Lys447) in the Fc region may be present or may not be present. In one aspect, a heavy chain comprising an Fc region as specified herein is included in an antibody according to the invention, the heavy chain comprising an additional C-terminal glycine-lysine dipeptide (G446 and K447, EU numbering system). In one aspect, a heavy chain comprising an Fc region as specified herein is included in an antibody according to the invention, the heavy chain comprising an additional C-terminal glycine residue (G446, numbered according to the EU index). Unless otherwise provided herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, which is also referred to as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, U.S. Department of Health and Human Services, National Institutes of Health, Bethesda, MD, 1991.

[0183] "Framework" or "FR" refers to the variable domain residues excluding the complementarity determining regions (CDRs). The FR of a variable domain is usually composed of the following four FR domains: FR1, FR2, FR3, and FR4. Therefore, CDR and FR sequences usually appear in the following sequence in VH (or VL): FR1-CDR-H1 (CDR-L1)-FR2-CDR-H2 (CDR-L2)-FR3-CDR-H3 (CDR-L3)-FR4.

[0184] The terms "full length antibody", "intact antibody" and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to a native antibody structure or having a heavy chain containing an Fc region as defined herein. It should be understood that a full length antibody comprises a heavy chain variable domain and a light chain variable domain as defined herein, and an Fc region as defined herein.

[0185] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and progeny derived from the primary transformed cell, regardless of the number of passages. Progeny may not be identical to the nucleic acid content of the parent cell, but may contain mutations. Mutant progeny as screened or selected for the same function or biological activity as in the original transformed cell are included herein.

[0186] A "human antibody" is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or derived from a non-human antibody using a full repertoire of human antibodies or other human antibody coding sequences. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues.

[0187] A "human consensus framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. In general, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. In general, the subgroup of sequences is a subgroup as described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), Volumes 1-3. In one aspect, for VL, the subgroup is subgroup κI as described in Kabat et al. in the above-mentioned document. In one aspect, for VH, the subgroup is subgroup III as described in Kabat et al. in the above-mentioned document.

[0188] "Humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human CDRs and amino acid residues from human FRs. In some aspects, a humanized antibody will substantially comprise at least one and typically two variable domains, wherein all or substantially all CDRs correspond to those of non-human antibodies, and all or substantially all FRs correspond to those of human antibodies. A humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. An antibody in "humanized form", such as a non-human antibody, refers to an antibody that has been humanized.

[0189] As used herein, the term "hypervariable region" or "HVR" refers to individual regions of an antibody variable domain that are hypervariable in sequence and determine antigen binding specificity, such as the "complementarity determining region" ("CDR").

[0190] In some aspects, the antibody comprises six CDRs; three in VH (CDR-H1, CDR-H2, CDR-H3), and three in VL (CDR-L1, CDR-L2, CDR-L3). In some aspects, the antibody comprising six CDRs is a full-length antibody. In some aspects, the antibody comprising six CDRs is an antibody fragment.

[0191] Exemplary CDRs herein include:

[0192] (a) hypervariable loops occurring at the following amino acid residues: 26 to 32 (L1), 50 to 52 (L2), 91 to 96 (L3), 26 to 32 (H1), 53 to 55 (H2), and 96 to 101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987));

[0193] (b) CDRs occurring at amino acid residues 24 to 34 (L1), 50 to 56 (L2), 89 to 97 (L3), 31 to 35b (H1), 50 to 65 (H2), and 95 to 102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and

[0194] (c) Antigenic contact points occurring at amino acid residues 27c to 36 (L1), 46 to 55 (L2), 89 to 96 (L3), 30 to 35b (H1), 47 to 58 (H2), and 93 to 101 (H3) (MacCallum et al. J. Mol. Biol. 262:732-745 (1996)).

[0195] Unless otherwise indicated, CDRs are identified as described in Kabat et al., supra and Chothia, supra. One skilled in the art will appreciate that CDR names may also be identified according to McCallum, supra or any other scientifically accepted nomenclature system.

[0196] In one aspect, the CDR residues comprise Figures 5A to 5D and Figures 6A to 6D and those identified in Tables C1, C2, D1 and D2. In other aspects, the CDR residues comprise those identified in Tables N1, N2, O1 and O2.

[0197] A "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain aspects, the subject is a human. In some aspects, the subject is a patient.

[0198] An "isolated" antibody is one that has been separated from a component of its natural environment. In some aspects, the antibody is purified to a purity greater than 95% or 99% by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0199] The term "nucleic acid molecule" or "polynucleotide" includes any compound and / or substance comprising a nucleotide polymer. Each nucleotide is composed of a base, particularly a purine or pyrimidine base (i.e., cytosine (C), guanine (G), adenine (A), thymine (T) or uracil (U)), a sugar (i.e., deoxyribose or ribose) and a phosphate group. Typically, nucleic acid molecules are described by a base sequence, wherein the base represents the primary structure (linear structure) of the nucleic acid molecule. Typically, the sequence of bases is represented from 5' to 3'. In this article, the term nucleic acid molecule encompasses synthetic forms of deoxyribonucleic acid (DNA) (including, for example, complementary DNA (cDNA) and genomic DNA), ribonucleic acid (RNA) (particularly messenger RNA (mRNA)), DNA or RNA, and mixed polymers comprising two or more of these molecules. Nucleic acid molecules can be linear or cyclic. In addition, the term nucleic acid molecule includes sense strands and antisense strands, as well as single-stranded and double-stranded forms. In addition, nucleic acid molecules described herein may contain naturally occurring or non-naturally occurring nucleotides. Examples of non-naturally occurring nucleotides include modified nucleotide bases with derivatized sugar or phosphate backbone bonds or chemically modified residues. Nucleic acid molecules also encompass DNA and RNA molecules suitable as carriers for direct expression in vitro and / or in vivo (e.g., in a host or subject) of antibodies described herein. Such DNA (e.g., cDNA) or RNA (e.g., mRNA) vectors may be unmodified or modified. For example, mRNA may be chemically modified to enhance the stability of the RNA vector and / or the expression of the encoding molecule so that mRNA may be injected into a subject to produce antibodies in vivo (see, e.g., Stadler et al., Nature Medicine 2017, published online on June 12, 2017, doi: 10.1038 / nm.4356 or EP 2 101 823 B1).

[0200] An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0201] "Isolated nucleic acid encoding an anti-CCR8 antibody" refers to one or more nucleic acid molecules encoding the anti-CCR8 antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules in a single vector or separate vectors, and such nucleic acid molecules present in one or more locations in a host cell.

[0202] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, that is, except for possible variant antibodies (e.g., containing naturally occurring mutations or produced during the production of monoclonal antibody preparations, such variants are usually present in small amounts), each antibody comprising the population is identical and / or binds to the same epitope. Contrary to polyclonal antibody preparations that typically include different antibodies for different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed to a single determinant on an antigen. Therefore, the modifier "monoclonal" indicates that the characteristic of an antibody is obtained from a substantially homogeneous antibody population, and should not be interpreted as requiring antibodies to be produced by any particular method. For example, monoclonal antibodies according to the present disclosure can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of a human immunoglobulin locus, such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

[0203] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or a radiolabel. Naked antibodies can be present in pharmaceutical compositions.

[0204] "Native antibodies" refer to naturally occurring immunoglobulin molecules with different structures. For example, natural IgG antibodies are heterotetrameric glycoproteins of about 150,000 daltons, consisting of two identical light chains and two identical heavy chains bonded by disulfide bonds. From the N-terminus to the C-terminus, each heavy chain has a variable domain (VH), also referred to as a variable heavy chain domain or a heavy chain variable domain, followed by three constant heavy chain domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable domain (VL), also referred to as a variable light chain domain or a light chain variable domain, followed by a constant light chain (CL) domain.

[0205] The term "package insert" is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and / or warnings concerning the use of such therapeutic products.

[0206] "Amino acid sequence identity percentage (%)" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence after aligning the candidate sequence with the reference polypeptide sequence and introducing spaces (if necessary) to achieve maximum sequence identity percentage, and for the purpose of alignment without considering any conservative substitutions as components of sequence identity. Alignment for determining amino acid sequence identity percentage can be achieved in various ways within the skill of the art, for example using publicly available computer software such as BLAST, BLAST-2, Clustal W, Megalign (DNASTAR) software or FASTA program packages. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the compared sequence. Alternatively, the sequence comparison computer program ALIGN-2 can be used to generate identity percentage values. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc. and the source code has been filed with user documentation in the US Copyright Office, Washington DC, 20559, where it is registered under US Copyright Registration No. TXU510087 and is described in WO 2001 / 007611.

[0207] Unless otherwise indicated, for the purposes of this article, the values ​​of amino acid sequence identity percentages are generated using the BLOSUM50 comparison matrix using the ggsearch program of the FASTA package version 36.3.8c or higher. The FASTA program package is by W.R. Pearson and DJ Lipman (1988), "Improved Tools for Biological Sequence Analysis", PNAS 85:2444-2448; W.R. Pearson (1996) "Effective protein sequence comparison" Meth. Enzymol. 266:227-258; and Pearson et al. (1997) Genomics 46:24-36 creation and can be publicly obtained from fasta.bioch.virginia.edu / fasta_www2 / fasta_down.shtml or ebi.ac.uk / Tools / sss / fasta. Alternatively, sequences can be compared using a public server accessible at fasta.bioch.virginia.edu / fasta_www2 / index.cgi, using the ggsearch (global protein:protein) program and default options (BLOSUM50; open: -10; ext: -2; Ktup = 2) to ensure that a global rather than a local alignment is performed. The amino acid identity percentage is given in the output alignment header.

[0208] The terms "pharmaceutical composition" and "pharmaceutical formulation" are used interchangeably herein and refer to a preparation that is in a form that permits the biological activity of the active ingredients contained therein to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the formulation would be administered.

[0209] "Pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical composition or formulation other than the active ingredient, which is non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0210] Unless otherwise indicated, as used herein, the term "CCR8" refers to any natural CCR8 from any vertebrate source, including mammals such as primates (e.g., humans, monkeys ("cynomolgus monkeys")) and rodents (e.g., mice and rats). The term includes "full-length" unprocessed CCR8, as well as any form of CCR8 produced by processing in cells. The term also encompasses naturally occurring variants of CCR8, such as splice variants or allelic variants. In some aspects, CCR8 is human CCR8 ("hCCR8" or "huCCR8"). The amino acid sequence of exemplary human CCR8 is shown in SEQ ID NO: 106, as shown in the table below. In some aspects, CCR8 is cynomolgus monkey ("cyno") CCR8. The amino acid sequence of exemplary cynomolgus monkey CCR8 is shown in SEQ ID NO: 107, as shown in the table below. In some aspects, CCR8 is mouse CCR8 ("mCCR8"). The amino acid sequence of an exemplary mouse CCR8 is shown in SEQ ID NO: 108, as shown in the table below.

[0211]

[0212]

[0213] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners to eliminate T cell dysfunction caused by signaling on the PD-1 signaling axis, resulting in restoration or enhancement of T cell function (e.g., proliferation, cytokine production, and / or target cell killing). As used herein, PD-1 axis binding antagonists include PD-L1 binding antagonists, PD-1 binding antagonists, and PD-L2 binding antagonists. In some cases, PD-1 axis binding antagonists include PD-L1 binding antagonists or PD-1 binding antagonists. In a preferred aspect, the PD-1 axis binding antagonist is a PD-L1 binding antagonist.

[0214] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the signal transduction generated by the interaction of PD-L1 with one or more of its binding partners (such as PD-1 and / or B7-1). In some cases, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In a specific aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some cases, the PD-L1 binding antagonist includes anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with the signal transduction generated by the interaction of PD-L1 with one or more of its binding partners (such as PD-1 and / or B7-1). In one instance, the PD-L1 binding antagonist reduces the negative co-stimulatory signal mediated by signaling through PD-L1 mediated by or through cell surface proteins expressed on T lymphocytes, thereby rendering the dysfunctional T cells less dysfunctional (e.g., improving the response of effectors to antigen recognition). In some instances, the PD-L1 binding antagonist binds to PD-L1. In some instances, the PD-L1 binding antagonist is an anti-PD-L1 antibody (e.g., an anti-PD-L1 antagonist antibody). Exemplary anti-PD-L1 antagonist antibodies include atezolizumab, MDX-1105, MEDI4736 (durvalumab), MSB0010718C (avelumab), SHR-1316, CS1001, envafolimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001 , KL-A167, APL-502, cosibelimab, lodapolimab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, and HS-636. In some aspects, the anti-PD-L1 antibody is atezolizumab, MDX-1105, MEDI4736 (durvalumab), or MSB0010718C (avetumab). In one specific aspect, the PD-L1 binding antagonist is MDX-1105. In another specific aspect, the PD-L1 binding antagonist is MEDI4736 (durvalumab). In another specific aspect, the PD-L1 binding antagonist is MSB0010718C (avelumab).In other aspects, the PD-L1 binding antagonist can be a small molecule, for example, GS-4224, INCB086550, MAX-10181, INCB090244, CA-170 or ABSK041, which can be administered orally in some cases. Other exemplary PD-L1 binding antagonists include AVA-004, MT-6035, VXM10, LYN192, GB7003 and JS-003. In one aspect, the PD-L1 binding antagonist is atezolizumab.

[0215] For the purposes of this article, "atezumab" is an Fc-engineered, humanized, non-glycosylated IgG1 kappa immunoglobulin that binds to PD-L1. Atezolizumab contains a single amino acid substitution (asparagine to alanine) (N297A) at position 297 on the heavy chain, using EU numbering of Fc region amino acid residues, which results in a non-glycosylated antibody with minimal binding to Fc receptors. Atezolizumab is also described in WHO Drug Information (International Nonproprietary Drug Name (Proposed INN)) List 112, Vol. 28, No. 4, 2014, p. 488.

[0216] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with the signal transduction generated by the interaction of PD-1 with one or more of its binding partners (such as PD-L1 and / or PD-L2). PD-1 (programmed death 1) is also referred to as "programmed cell death 1", "PDCD1", "CD279" and "SLEB2" in the art. Exemplary human PD-1 is shown in UniProtKB / Swiss-Prot accession number Q15116. In some cases, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In a specific aspect, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies and antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with the signal transduction generated by the interaction of PD-1 with PD-L1 and / or PD-L2. In one instance, a PD-1 binding antagonist reduces negative co-stimulatory signals mediated by signaling through PD-1 mediated by or through cell surface proteins expressed on T lymphocytes, thereby rendering dysfunctional T cells less dysfunctional (e.g., improving effector responses to antigen recognition). In some instances, a PD-1 binding antagonist binds to PD-1. In some instances, a PD-1 binding antagonist is an anti-PD-1 antibody (e.g., an anti-PD-1 antagonist antibody). Exemplary anti-PD-1 antagonist antibodies include nivolumab, pembrolizumab, MEDI-0680, PDR001 (spartalizumab), REGN2810 (cemiprilimab), BGB-108, palolizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, dotalimumab, rivulimab, sazanlimab, penampalimab, CS1003, HLX10, SCT-I10A, sepalizumab, batilizumab, genoluzumab, BI 754091, silimumab, YBL-006, BAT1306, HX008, brigalimumab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103 and hAb21. In one specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In another specific aspect, the PD-1 binding antagonist is a PD-L2 fusion protein, for example, AMP-224. In another specific aspect, the PD-1 binding antagonist is MED1-0680.In another specific aspect, the PD-1 binding antagonist is PDR001 (spartalizumab). In another specific aspect, the PD-1 binding antagonist is REGN2810 (simiprilimab). In another specific aspect, the PD-1 binding antagonist is BGB-108. In another specific aspect, the PD-1 binding antagonist is palolizumab. In another specific aspect, the PD-1 binding antagonist is carrelizumab. In another specific aspect, the PD-1 binding antagonist is sindilimab. In another specific aspect, the PD-1 binding antagonist is tislelizumab. In another specific aspect, the PD-1 binding antagonist is toripalizumab. Other exemplary PD-1 binding antagonists include BION-004, CB201, AUNP-012, ADG104, and LBL-006.

[0217] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to an attempt to alter the natural course of a disease (e.g., cancer (e.g., locally advanced, recurrent or metastatic solid tumor malignancies)) in the subject being treated, and may be performed for prevention ("prophylactic treatment" or "prophylactic treatment") or may be performed during the course of clinical pathology ("therapeutic treatment" or "therapeutic treatment"). Desired effects of therapeutic treatment include, but are not limited to, alleviation of symptoms, attenuation of any direct or indirect pathological consequences of the disease, prevention of metastasis of cancer, reduction of the rate of disease progression, amelioration or palliation of the disease state, and alleviation or improved prognosis. Desired effects of prophylactic treatment include, but are not limited to, prevention of the occurrence or recurrence of the disease. In some aspects, the antibodies described herein are used to delay the development of a disease or slow the progression of a disease.

[0218] The term "variable region" or "variable domain" refers to the domain of an antibody heavy chain or light chain that participates in the binding of an antibody to an antigen. The variable domains of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have similar structures, wherein each domain comprises four conserved framework regions (FRs) and three complementary determining regions (CDRs). (See, e.g., Kindt et al. Kuby Immunology, 6th edition, WH Freeman and Co., p. 91 (2007).) A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen may be isolated using VH or VL domains from antibodies that bind to the antigen, respectively, to screen for libraries of complementary VL or VH domains. See, e.g., Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

[0219] The term "vector" as used herein refers to a nucleic acid molecule capable of carrying another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures, as well as vectors that are incorporated into the genome of a host cell into which they have been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors."

[0220] II. Methods and Compositions

[0221] In one aspect, the present disclosure is based, in part, on the development of therapeutic methods and dosing regimens for treating locally advanced, recurrent, or metastatic solid tumor malignancies using an anti-CCR8 antibody (e.g., an anti-CCR8 antibody as disclosed herein).

[0222] A. Methods of Treatment and Compositions for Use

[0223] The present disclosure provides therapeutic methods and compositions for use in treating cancer (e.g., locally advanced, recurrent or metastatic solid tumor malignancies) in a subject in need thereof, which may include administering to the subject an anti-CCR8 antibody as disclosed herein alone or in combination with one or more additional therapeutic agents (e.g., a PD-1 axis binding antagonist, e.g., an anti-PD-L1 antibody such as atezolizumab). Any anti-CCR8 antibody (e.g., a monoclonal antibody that binds to CCR8) may be used.

[0224] In one aspect, provided herein is a method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8.

[0225] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

[0226] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

[0227] In one aspect, provided herein is a method of depleting regulatory T cells ("Tregs") in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8.

[0228] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

[0229] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

[0230] In some aspects, (i) the subject has progressed after at least one available standard therapy; and / or (ii) the subject is a subject for whom all available standard therapies have proven ineffective or are intolerant or contraindicated.

[0231] In some aspects, locally advanced, recurrent or metastatic solid tumors are incurable.

[0232] In some aspects, the subject is 18 years of age or older. For example, the subject can be an adult.

[0233] Any suitable locally advanced, recurrent or metastatic solid tumor malignancy can be treated. For example, in some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), melanoma, triple negative breast cancer (TNBC), urothelial carcinoma (UC), esophageal cancer, gastric cancer, cervical cancer, renal cell carcinoma (RCC) or hepatocellular carcinoma (HCC).

[0234] In some aspects, the RCC is clear cell RCC.

[0235] In some aspects, the HNSCC is HNSCC of the oral cavity, oropharynx, hypopharynx, or larynx.

[0236] In some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is NSCLC.

[0237] In some aspects, the subject's tumor contains a targetable somatic alteration, and the subject has experienced disease progression during or after treatment with the targeted agent or is intolerant to the treatment.

[0238] In some aspects, targetable somatic alterations include somatic alterations involving epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), ROS proto-oncogene 1 (ROS1), proto-oncogene B-Raf (BRAF) V600E, neurotrophic tyrosine receptor kinase (NTRK), MET proto-oncogene (MET), RET proto-oncogene (RET), or Kirsten rat sarcoma virus (KRAS).

[0239] In some aspects, the melanoma is cutaneous melanoma.

[0240] In some aspects, the subject's tumor comprises a BRAFV600 mutation and the subject has experienced disease progression during or after treatment with or is intolerant to treatment with one or more serine / threonine protein kinase B-Raf (BRAF) inhibitors and / or one or more mitogen-activated protein kinase (MEK) inhibitors.

[0241] In some aspects, the locally advanced, recurrent, or metastatic solid tumor malignancy is UC.

[0242] In some aspects, the subject has: (i) histologically confirmed incurable advanced transitional cell carcinoma of the urothelium (including the renal pelvis, ureters, bladder, and urethra); and / or (ii) mixed histology, in which the subject's tumor has a dominant transitional cell pattern.

[0243] In some aspects, the locally advanced, recurrent or metastatic solid tumor malignancy is TNBC.

[0244] In some aspects, TNBC is defined by the American Society of Clinical Oncology-College of American Pathologists guidelines: (i) <1% of tumor cell nuclei are immunoreactive for estrogen receptor and <1% of tumor cell nuclei are immunoreactive for progesterone receptor; and / or (ii) HER2 negativity based on immunohistochemistry (IHC) and / or in situ hybridization.

[0245] In some aspects, the subject is checkpoint inhibitor (CPI) naive.

[0246] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is NSCLC or UC.

[0247] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is UC, wherein the subject is eligible for treatment with cisplatin, and the subject has experienced disease progression during or after treatment with cisplatin or is intolerant to the treatment.

[0248] In some aspects, the subject has not received prior treatment with a CPI, or where the subject has received adjuvant therapy with a CPI, the adjuvant therapy has been discontinued at least six months prior to the first administration of a monoclonal antibody that binds to CCR8 to the subject.

[0249] In some aspects, the subject has experienced a CPI.

[0250] In some aspects, the subject's locally advanced, recurrent, or metastatic solid tumor malignancy is NSCLC, HNSCC, melanoma, UC, TNBC, esophageal cancer, gastric cancer, cervical cancer, clear cell RCC, or HCC.

[0251] In some aspects, the subject receives clinical benefit from treatment comprising a PD-1 axis binding antagonist prior to disease progression.

[0252] In some aspects, the PD-1 axis binding antagonist is an anti-PD-L1 antibody or an anti-PD-1 antibody.

[0253] In some aspects, the subject has a treatment duration of greater than or equal to 6 months with a therapy comprising a PD-1 axis binding antagonist and / or has a partial response or a complete response as the best objective response.

[0254] In some aspects, (i) the subject has not been treated with a CPI, an immunomodulatory monoclonal antibody, or an immunomodulatory monoclonal antibody-derived therapy within 6 weeks prior to the first administration of a monoclonal antibody that binds to CCR8 to the subject; or (ii) the subject has previously been treated with a PD-1 axis binding antagonist, and the last administration of the PD-1 axis binding antagonist to the subject was at least 3 weeks prior to the first administration of the monoclonal antibody that binds to CCR8 to the subject.

[0255] In some aspects, the CPI is a PD-1 axis binding antagonist or a CTLA4 antagonist (e.g., an anti-CTLA4 antibody such as ipilimumab ).

[0256] In some aspects, the PD-1 axis binding antagonist is an anti-PD-L1 antibody (e.g., any anti-PD-L1 antibody disclosed herein, such as atezolizumab or avelumab) or an anti-PD-1 antibody (e.g., any anti-PD-1 antibody disclosed herein, such as pembrolizumab).

[0257] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject in a dosing regimen that includes one or more dosing cycles.

[0258] In some aspects, the one or more dosing cycles include a 21 day dosing cycle.

[0259] For example, in one aspect, provided herein is a method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 in a dosing regimen comprising one or more 21-day dosing cycles.

[0260] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof with a dosing regimen comprising one or more 21-day dosing cycles.

[0261] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof with a dosing regimen comprising one or more 21-day dosing cycles.

[0262] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject on day 1 of each 21 -day dosing cycle.

[0263] In one aspect, provided herein is a method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject every three weeks (Q3W) a monoclonal antibody that binds to CCR8.

[0264] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in the treatment of locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof every three weeks (Q3W).

[0265] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for the every three weeks (Q3W) treatment of locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof.

[0266] In one aspect, provided herein is a method of depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 every three weeks (Q3W).

[0267] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use every three weeks (Q3W) in depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

[0268] In one aspect, provided herein is a method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg.

[0269] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof at a dose of 2 mg.

[0270] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof at a dose of 2 mg.

[0271] In one aspect, provided herein is a method of depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg.

[0272] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof at a dose of 2 mg.

[0273] In one aspect, provided herein is a method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg every three weeks (Q3W).

[0274] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof at a dose of 2 mg every three weeks (Q3W).

[0275] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof at a dose of 2 mg every three weeks (Q3W).

[0276] In one aspect, provided herein is a method for depleting Tregs in the tumor microenvironment of a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg every three weeks (Q3W).

[0277] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use at a dose of 2 mg every three weeks (Q3W) to deplete Tregs in the tumor microenvironment of locally advanced, recurrent or metastatic solid tumor malignancies in a subject in need thereof.

[0278] In some embodiments of any of the aforementioned aspects, a monoclonal antibody that binds to CCR8 can be administered to a subject until disease progression or unacceptable toxicity occurs.

[0279] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject at a dose of 2 mg.

[0280] Any suitable route of administration can be used in the methods and compositions for use disclosed herein.In some aspects, the monoclonal antibody that binds to CCR8 is administered intravenously to the subject.

[0281] In some aspects, the monoclonal antibody that binds to CCR8 is administered to the subject intravenously by infusion.

[0282] In some aspects, a monoclonal antibody that binds to CCR8 is administered to a subject as a monotherapy.

[0283] In other aspects, the monoclonal antibody combined with CCR8 is administered to the subject as a combination therapy. For example, the combination therapy may include administering an antibody as described herein and administering at least one additional therapeutic agent (e.g., one, two, three, four, five or six additional therapeutic agents).

[0284] The one or more additional therapeutic agents encompass any medicament that can be administered for treatment. In some aspects, the additional therapeutic agent is another anticancer agent. Exemplary anticancer agents include, but are not limited to, microtubule disruptors, antimetabolites, topoisomerase inhibitors, DNA intercalators, alkylating agents, hormone therapy, kinase inhibitors, receptor antagonists, tumor cell apoptosis activators, anti-angiogenic agents, immunomodulators, cell adhesion inhibitors, cytotoxic agents or cell inhibitors, apoptosis activators, agents that increase cell sensitivity to apoptosis inducing agents, cytokines, anticancer vaccines or oncolytic viruses, Toll-like receptors (TLR) agents, bispecific antibodies, cell therapy, and immune cell conjugates. In certain aspects, the additional therapeutic agent is an immunomodulatory anti-cancer agent, e.g., a checkpoint inhibitor (CPI) such as an anti-CTLA4 antibody (e.g., ipilimumab) or a PD-1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody (e.g., atezolizumab or avelumab), a PD-1 binding antagonist (e.g., pembrolizumab) or a PD-L2 binding antagonist).

[0285] In some aspects, the one or more additional therapeutic agents include an anti-PD-L1 antibody. For example, in some aspects, the one or more additional therapeutic agents include atezolizumab.

[0286] In some aspects, atezolizumab is administered to a subject in a dosing regimen comprising one or more dosing cycles. In some aspects, the one or more dosing cycles comprise a 14-day, 21-day, or 28-day dosing cycle.

[0287] In some aspects, the one or more dosing cycles include a 21-day dosing cycle. In some aspects, atezolizumab is administered to the subject on day 1 of each 21-day dosing cycle.

[0288] Any suitable dose of atezolizumab may be administered to a subject. In some aspects, atezolizumab is administered to a subject at a dosage of 1200 mg. For example, in some aspects, atezolizumab is administered to a subject at a dosage of 1200 mg every three weeks (Q3W). In other examples, atezolizumab is administered to a subject at a dosage of 840 mg (e.g., every two weeks (Q2W)). In yet another example, atezolizumab is administered to a subject at a dosage of 1680 mg (e.g., every four weeks (Q4W)).

[0289] In some aspects, atezolizumab is administered intravenously to the subject. In some aspects, atezolizumab is administered intravenously to the subject by infusion.

[0290] In some aspects, it has been determined that a tumor sample from a subject has a detectable PD-L1 expression level. For example, in some aspects, a tumor sample from a subject has greater than or equal to 1% of tumor cells (TC), immune cells (IC), a comprehensive positive score (CPS) or a tumor proportion score (TPS). The presence or expression level of PD-L1 can be determined using any suitable method (e.g., using immunohistochemistry of anti-PD-L1 diagnostic antibodies). Any suitable anti-PD-L1 diagnostic antibody can be used, for example, SP142 (VENTANA), SP263 (VENTANA), 22C3 (Dako), 28-8 (Dako), E1L3N, 4059, h5H1, 9A11, etc.

[0291] In some aspects, the subject has received at least two cycles (e.g., at least two, three, four, five, six, seven, eight, nine, ten, or more cycles) of a monoclonal antibody that binds to CCR8 prior to administering atezolizumab to the subject.

[0292] Such combination therapies described above encompass combined administration (where two or more therapeutic agents are included in the same or separate pharmaceutical compositions) and separate administration, in which case the administration of an antibody as described herein may be performed before, simultaneously with, and / or after the administration of an additional therapeutic agent or medicament. In one aspect, the administration of an anti-CCR8 antibody and the administration of an additional therapeutic agent are performed within about one month of each other, or within about one week, two weeks, or three weeks, or within about one day, two days, three days, four days, five days, or six days. In one aspect, the antibody and the additional therapeutic agent are administered to the subject on day 1 of treatment. The antibodies as described herein may also be used in combination with radiation therapy.

[0293] In one aspect, an anti-CCR8 antibody for use as a medicine is provided. In a further aspect, an anti-CCR8 antibody for treating cancer is provided. In certain aspects, an anti-CCR8 antibody for use in a method of treating a subject (e.g., a human subject) in need thereof is provided. The method comprises administering an effective amount of an anti-CCR8 antibody to the subject. In one such aspect, such as described below, the method further comprises administering an effective amount of at least one additional therapeutic agent (e.g., one, two, three, four, five or six additional therapeutic agents) to the subject. In other aspects, the present disclosure provides an anti-CCR8 antibody for depleting Tregs in a tumor microenvironment. In certain aspects, the present disclosure provides an anti-CCR8 antibody for depleting Tregs in a subject's tumor microenvironment, the method comprising administering an effective amount of an anti-CCR8 antibody to the subject to deplete Tregs in the tumor microenvironment.

[0294] In a further aspect, the present disclosure provides the use of anti-CCR8 antibodies in the manufacture or preparation of a medicament. In one aspect, the medicament is used to treat cancer. In a further aspect, the medicament is used in a method for treating cancer, the method comprising administering an effective amount of the medicament to a subject (e.g., a human subject) in need thereof. In one such aspect, for example as described below, the method further comprises administering an effective amount of at least one additional therapeutic agent to the subject. In a further aspect, the medicament is used to deplete Tregs in a tumor microenvironment. In a further aspect, the medicament is used in a method for depleting Tregs in a tumor microenvironment of a subject, the method comprising administering an effective amount of the medicament to the subject to eliminate Tregs in the tumor microenvironment.

[0295] In a further aspect, the present disclosure provides a method for treating cancer. In one aspect, the method comprises administering an effective amount of an anti-CCR8 antibody to a subject (e.g., a human subject) in need thereof to treat cancer. In one such aspect, as described below, the method further comprises administering an effective amount of at least one additional therapeutic agent to the subject.

[0296] In a further aspect, the present disclosure provides an anti-CCR8 antibody for depleting, for example, Treg cells outside or in a tumor microenvironment. For example, in certain embodiments, the present disclosure provides a method for depleting Treg cells in a tumor microenvironment of a subject (e.g., a human subject) in need thereof, the method comprising administering to the subject an effective amount of an anti-CCR8 antibody sufficient to deplete Treg cells in the tumor microenvironment, thereby treating cancer. In some aspects, the present disclosure provides a method for depleting Treg cells outside (e.g., in circulation) of a subject (e.g., a human subject) in need thereof, the method comprising administering to the subject an effective amount of an anti-CCR8 antibody sufficient to deplete Treg cells outside the tumor microenvironment, thereby treating cancer. Without wishing to be bound by any particular theory, by reducing the number of Treg cells outside the tumor microenvironment, as the number of Treg cells infiltrating into the tumor microenvironment decreases, the number of Treg cells in the tumor microenvironment is reduced.

[0297] In one aspect, provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject every three weeks (Q3W) a monoclonal antibody that binds to CCR8.

[0298] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in the every three weeks (Q3W) treatment of cancer in a subject in need thereof.

[0299] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for the every three weeks (Q3W) treatment of cancer in a subject in need thereof.

[0300] In one aspect, provided herein is a method of depleting Tregs in a tumor microenvironment of a cancer in a subject in need thereof, the method comprising administering to the subject every three weeks (Q3W) a monoclonal antibody that binds to CCR8.

[0301] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in depleting Tregs in the tumor microenvironment of a cancer in a subject in need thereof every three weeks (Q3W).

[0302] In one aspect, provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg.

[0303] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating cancer in a subject in need thereof at a dose of 2 mg.

[0304] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating cancer in a subject in need thereof at a dose of 2 mg.

[0305] In one aspect, provided herein is a method of depleting Tregs in a tumor microenvironment of a cancer in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg.

[0306] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in depleting Tregs in a tumor microenvironment of a cancer in a subject in need thereof at a dose of 2 mg.

[0307] In one aspect, provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg every three weeks (Q3W).

[0308] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in treating cancer in a subject in need thereof at a dose of 2 mg every three weeks (Q3W).

[0309] In another aspect, provided herein is the use of a monoclonal antibody that binds to CCR8 in the manufacture of a medicament for treating cancer in a subject in need thereof at a dose of 2 mg every three weeks (Q3W).

[0310] In one aspect, provided herein is a method of depleting Tregs in a tumor microenvironment of a cancer in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CCR8 at a dose of 2 mg every three weeks (Q3W).

[0311] In another aspect, provided herein is a monoclonal antibody that binds to CCR8 for use in depleting Tregs in the tumor microenvironment of a cancer in a subject in need thereof at a dose of 2 mg every three weeks (Q3W).

[0312] Exemplary cancers include, but are not limited to, bladder cancer (e.g., urothelial carcinoma), blastoma, blood cancer (e.g., lymphoma such as non-Hodgkin's disease, leukemia), bone cancer, brain cancer, breast cancer (e.g., triple-negative breast cancer), cervical cancer, colorectal cancer (e.g., colon cancer, rectal cancer), endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma), kidney cancer (e.g., renal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer, small cell lung cancer), ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer (e.g., melanoma, squamous cell carcinoma, cell carcinoma), testicular cancer, and uterine cancer.

[0313] In certain aspects, the cancer is bladder cancer, blood cancer, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, stomach cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, and skin cancer.

[0314] In certain aspects, the cancer is bladder cancer, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, head and neck cancer, liver cancer, lung cancer, or skin cancer.

[0315] In certain aspects, the cancer is a solid tumor, eg, a locally advanced or metastatic solid tumor.

[0316] In certain aspects, the cancer (eg, a locally advanced, recurrent, or metastatic solid tumor malignancy) expresses CCR8.

[0317] In certain aspects, the cancer (e.g., a locally advanced, recurrent, or metastatic solid tumor malignancy) is a T cell inflamed tumor or comprises a T cell inflamed tumor microenvironment.

[0318] In certain aspects, cancer (e.g., locally advanced, recurrent or metastatic solid tumor malignancies) includes regulatory T cells in the tumor microenvironment, and as described herein, exposing the cancer to CCR8 antibodies results in depletion of regulatory T cells in the tumor microenvironment. In a further aspect, the disclosure provides a pharmaceutical composition comprising any of the anti-CCR8 antibodies described herein, for example, for use in any of the above methods of treatment. In one aspect, a pharmaceutical composition comprising any of the anti-CCR8 antibodies provided herein, and a pharmaceutically acceptable carrier. In another aspect, a pharmaceutical composition comprising any of the anti-CCR8 antibodies provided herein, and at least one additional therapeutic agent, for example, as described below.

[0319] Any of the anti-CCR8 antibodies provided herein (e.g., in Section B below) can be used in the therapeutic methods and compositions for use (e.g., anti-CCR8 antibodies for use (e.g., monoclonal antibodies that bind to CCR8 for use)) as disclosed herein.

[0320] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:29 or SEQ ID NO:30, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:31, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:32; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:26, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:27, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:28.

[0321] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0322] In some aspects, the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:106.

[0323] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 to 52; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0324] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35-47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48-52.

[0325] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:48; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0326] In some aspects, the monoclonal antibody that binds to CCR8 comprises: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:47; and (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:48.

[0327] In some aspects, VL comprises a V4M mutation, a P43A mutation, a F46L mutation, a C90Q mutation, or a combination thereof. In some cases, the V4M mutation, the P43A mutation, the F46L mutation, or the C90Q mutation is numbered according to Kabat.

[0328] In some aspects, VH comprises a G49S mutation, a K71R mutation, a S73N mutation, or a combination thereof. In some cases, the G49S mutation, the K71R mutation, or the S73N mutation is numbered according to Kabat.

[0329] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:55 and the light chain amino acid sequence of SEQ ID NO:56.

[0330] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:60 and the light chain amino acid sequence of SEQ ID NO:56.

[0331] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:111 and the light chain amino acid sequence of SEQ ID NO:56.

[0332] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:113 and the light chain amino acid sequence of SEQ ID NO:56.

[0333] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35-47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48-52.

[0334] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:47 and the VL sequence of SEQ ID NO:48.

[0335] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5, (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; and a light chain variable domain (VL) comprising (d) CDR-L1 comprising the amino acid sequence of SEQ ID NO:1, (e) CDR-L2 comprising the amino acid sequence of SEQ ID NO:2, and (f) CDR-L3 comprising the amino acid sequence of SEQ ID NO:3.

[0336] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0337] In some aspects, the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:106.

[0338] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0339] In some aspects, the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 10-21 and a VL sequence selected from the group consisting of SEQ ID NOs: 22-25.

[0340] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:24; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0341] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:21; and a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:24.

[0342] In some aspects, the VL comprises a Y2I mutation. In some cases, the Y2I mutation is numbered according to Kabat.

[0343] In some aspects, VH comprises S73N mutation, V78L mutation, T76N mutation, F91Y mutation and P105Q mutation or its combination.In some cases, S73N mutation, V78L mutation, T76N mutation, F91Y mutation or P105Q mutation is according to Kabat numbering.

[0344] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:57 and the light chain amino acid sequence of SEQ ID NO:58.

[0345] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:61 and the light chain amino acid sequence of SEQ ID NO:58.

[0346] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:112 and the light chain amino acid sequence of SEQ ID NO:58.

[0347] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:114 and the light chain amino acid sequence of SEQ ID NO:58.

[0348] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:21 and the VL sequence of SEQ ID NO:24.

[0349] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 84, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 85; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 75.

[0350] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:95; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:94; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0351] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:95 and the VL sequence of SEQ ID NO:94.

[0352] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:101 and the light chain amino acid sequence of SEQ ID NO:100.

[0353] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:115 and a light chain amino acid sequence of SEQ ID NO:100.

[0354] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78.

[0355] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:97; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:96; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0356] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:97 and the VL sequence of SEQ ID NO:96.

[0357] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:103 and the light chain amino acid sequence of SEQ ID NO:102.

[0358] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:116 and a light chain amino acid sequence of SEQ ID NO:102.

[0359] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:90 or SEQ ID NO:91, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:92, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:93; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:79, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:80, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:81.

[0360] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:99; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:98; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0361] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:99 and the VL sequence of SEQ ID NO:98.

[0362] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:105 and a light chain amino acid sequence of SEQ ID NO:104.

[0363] In some aspects, the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO:117 and a light chain amino acid sequence of SEQ ID NO:104.

[0364] In some aspects, the monoclonal antibody that binds to CCR8 binds to CCR8 independent of sulfation of CCR8.

[0365] In some aspects, the antibody binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:106.

[0366] In some aspects, the antibody binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:106.

[0367] In some aspects, the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:65 or SEQ ID NO:66, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:67, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:68; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:62, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:63, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:64.

[0368] In some aspects, the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:70; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:69; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

[0369] In some aspects, the monoclonal antibody that binds to CCR8 comprises the VH sequence of SEQ ID NO:70 and the VL sequence of SEQ ID NO:69.

[0370] In some aspects, the monoclonal antibody that binds to CCR8 comprises the heavy chain amino acid sequence of SEQ ID NO:72 and the light chain amino acid sequence of SEQ ID NO:71.

[0371] In some aspects, the monoclonal antibody that binds to CCR8 is a human antibody.

[0372] In some aspects, the monoclonal antibody that binds to CCR8 is a humanized antibody.

[0373] In some aspects, the monoclonal antibody that binds to CCR8 is a chimeric antibody.

[0374] In some aspects, the monoclonal antibody that binds to CCR8 is an antibody fragment that binds to CCR8.

[0375] In some aspects, the monoclonal antibody that binds to CCR8 is a full-length antibody.

[0376] In some aspects, the monoclonal antibody that binds to CCR8 is a full-length IgG1 antibody.

[0377] In some aspects, the monoclonal antibody that binds to CCR8 comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO:53 or SEQ ID NO:59.

[0378] In some aspects, the monoclonal antibody that binds to CCR8 comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO:54.

[0379] In some aspects, the monoclonal antibody that binds to CCR8 is present at about 1×10 -12 M is about 1×10 -11 The binding affinity (K d ) binds to CCR8.

[0380] In some aspects, the CCR8 is human CCR8.

[0381] In some aspects, the monoclonal antibody that binds to CCR8 is afucosylated.

[0382] In some aspects, the defucosylation ratio is between about 80% and about 95%.

[0383] In some aspects, regulatory T cells present in the tumor microenvironment of locally advanced, recurrent or metastatic solid tumor malignancies are depleted.

[0384] In some aspects, regulatory T cells outside the tumor microenvironment of locally advanced, recurrent or metastatic solid tumor malignancies are depleted.

[0385] In some aspects, the subject is a human.

[0386] Antibodies as described herein (and any additional therapeutic agents) can be administered by any suitable means, including parenteral, intrapulmonary and intranasal, and if necessary for local treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. Administration can be carried out by any suitable route, for example by injection, such as intravenous or subcutaneous injection, depending in part on whether the administration is short-lived or long-term. Various dosing schedules are contemplated herein, including but not limited to single or multiple administrations, push administrations, and pulse infusions at various time points.

[0387] Antibodies as described herein will be formulated, dosed and applied in a manner consistent with good medical practice. Factors to be considered in this case include the specific condition being treated, the specific subject species being treated, the clinical condition of the subject, the cause of the condition, the delivery site of the medicament, the method of administration, the time schedule of administration, and other factors known to practitioners. The antibody is not necessary, but is optionally co-formulated with one or more preparations currently used to treat the condition in question. The effective amount of these other preparations depends on the amount of the antibody present in the pharmaceutical composition, the type of condition or treatment, and other factors discussed above. These are generally used with the same dosage and route of administration as described herein, or with about 1% to 99% of the dosage described herein, or with any dosage and any route determined to be suitable empirically / clinically.

[0388] The antibody is appropriately administered to the subject once or in a series of treatments. For repeated administration over several days or longer, depending on the condition, treatment will generally continue until the desired suppression of disease symptoms occurs. However, other dosage regimens may be available. The progress of the therapy can be monitored by conventional techniques and assays.

[0389] B. Exemplary Anti-CCR8 Antibodies

[0390] Any anti-CCR8 antibody can be used in any of the methods and compositions for use disclosed herein, eg, as described in Section A above.

[0391] In one aspect, the disclosure provides antibodies that bind to CCR8. In one aspect, the antibody provided is an isolated antibody that binds to CCR8. In one aspect, the disclosure provides antibodies that specifically bind to CCR8. In some aspects, anti-CCR8 antibodies bind to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:106. In some aspects, anti-CCR8 antibodies bind to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:106. In some aspects, CCR8 is human CCR8, mouse CCR8 or cynomolgus monkey CCR8. In some aspects, CCR8 is human CCR8. In one aspect, the disclosure provides antibodies that bind to CCR8 independently of tyrosine sulfation of CCR8 ("independent of sulfation"). Exemplary antibodies that rely on sulfation disclosed herein include Ab4 and Ab5, which will be further described in more detail below.

[0392] In certain aspects, the antibodies provided herein have a concentration 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 smaller, e.g. 10 -8 M to 10-13 M, for example 10 -9 M to 10 -13 The dissociation constant (K D ). In certain aspects, the antibody that binds to CCR8 has a specificity of about 1×10 -12 M is about 1×10 -10 M, about 1×10 -12 M is about 1×10 -11 M, or about 1×10 -11 M is about 5×10 -11 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 2×10 -11 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 5×10 -12 M of K d In one aspect, K D Radiolabeled IgG and a CHO cell line stably expressing the antigen were used for measurement. Stable CHO cells expressing the antigen were seeded at 50,000 cells per well in cold binding buffer (Opti-MEM + 2% fetal bovine serum (FBS) + 50 mM HEPES, pH 7.2 + 0.1% sodium azide). NEX244 Method (Perkin Elmer) fixed concentration of 125 I radiolabeled target antigen was mixed with serial dilutions of target antibody starting from 20nM or 50nM. The antibody mixture was added to the cells and incubated for 12 hours at room temperature under gentle agitation. The cells and antibodies were then transferred to a Millipore multi-layer screen filter plate. The filter plate was washed 4 times with 250 μL of cold binding buffer, dried for at least 30 minutes, and the filter was placed in a 5mL polystyrene tube. PerkinElmer Wallac Radioactivity was measured using a 2470 gamma counter set to 1 count per minute with a counting efficiency of 0.8. The data were fit to a heterologous single-site fit Ki competitive binding model.

[0393] In some aspects, the average clearance rate of the antibody provided herein after intravenous administration of a single 10mg / kg dose on the first day is about 3 to about 5mL / day / kg over a period of 35 days. For example, but not limited to, such administration may include the mAb of a single 10mg / kg IV push. Blood samples may be collected for analysis, for example, at 0.25 hours, 2 hours and 6 hours after administration, and 1 day, 2 days, 7 days, 14 days, 21 days, 28 days and 35 days, and a variety of means (e.g., qualified ELISA analytical methods) may be used to measure the mAb concentration in serum. In some aspects, administration is for mammals. In some aspects, administration is for primates. In some aspects, administration is for non-human primates, such as cynomolgus monkeys. In some aspects, administration is for people.

[0394] (i) Embodiments of Ab5 and its fragments

[0395] In one aspect, the present disclosure provides an anti-CCR8 antibody comprising at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 26; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 27; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 28. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to both human CCR8 and cynomolgus monkey CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0396] In one aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31; and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32. In one aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32. In another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32; and CDR-L3 comprising an amino acid sequence of SEQ ID NO: 28. In yet another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32; CDR-L3 comprising an amino acid sequence of SEQ ID NO: 28; and CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31. In yet another aspect, the antibody comprises: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:29 or SEQ ID NO:30; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:31; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:32.

[0397] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 28. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 28.

[0398] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30; (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31; and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 26; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 27; and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 28. In certain aspects, the anti-CCR8 antibody comprises all six of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to both human CCR8 and cynomolgus monkey CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0399] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:29 or SEQ ID NO:30, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:31, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:32; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:26, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:27, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:28.

[0400] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequences selected from the group consisting of SEQ ID NOs: 35 to 47. In another embodiment, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequences selected from the group consisting of SEQ ID NOs: 48 to 52. In another embodiment, the anti-CCR8 antibody comprises a CDR sequence of the VH sequence selected from the group consisting of SEQ ID NOs: 35 to 47 and a CDR sequence of the VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52.

[0401] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 47. In another embodiment, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 48. In another embodiment, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 47. In another embodiment, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 48.

[0402] In another aspect, the anti-CCR8 antibody comprises CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain selected from the group consisting of SEQ ID NOs: 35 to 47 and CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain selected from the group consisting of SEQ ID NOs: 48 to 52.

[0403] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:47 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:48.

[0404] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47 and a framework having at least 95% sequence identity with a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47. In another aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47 and a framework having at least 98% sequence identity with a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 35 to 47.

[0405] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 47 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 47. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 47 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 47. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 47 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 47. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 47; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 47.

[0406] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52 and a framework having at least 95% sequence identity with a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52. In another aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52 and a framework having (especially at least) at least 98% sequence identity with a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 48 to 52.

[0407] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 48 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 48. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 48 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 48. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 48 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 48. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 48; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 48.

[0408] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 31; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 32; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 28, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47; The amino acid sequences of the group consisting of SEQ ID NOs: 48 to 52 have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequences selected from the group consisting of SEQ ID NOs: 35 to 47. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequences selected from the group consisting of SEQ ID NOs: 48 to 52. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with an antibody comprising a VH sequence selected from the group consisting of SEQ ID NOs: 35 to 47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52 (K D ) is up to 10 times lower or up to 10 times higher.

[0409] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 31; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 32; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 28, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 47; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 47; NO:48 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:47. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:48. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 47 and the VL sequence of SEQ ID NO: 48 (K D ) is up to 10 times lower or up to 10 times higher.

[0410] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47. In one aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47. In certain aspects, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In some aspects, in the amino acid sequence selected from the group consisting of SEQ ID NO: 35 to 47, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In some aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises a VH sequence selected from the group consisting of SEQ ID NO: 35 to 47, including post-translational modifications of the sequence. In a specific aspect, VH comprises one, two or three CDRs selected from the following: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31, and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 48 to 52. In one aspect, the anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 48 to 52. In certain aspects, the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In some aspects, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence selected from the group consisting of SEQ ID NO: 48 to 52. In some aspects, the substitution, insertion or deletion occurs in a region outside of CDR (i.e., in FR).Optionally, the anti-CCR8 antibody comprises a VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52, including post-translational modifications of the sequence. In a specific aspect, the VL comprises one, two or three CDRs selected from the following: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 28.

[0411] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 47. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 47. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 47, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 47, including post-translational modifications of the sequence. In a specific aspect, the VH comprises one, two or three CDRs selected from the following: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 31, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 32. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 48. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 48. In some aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In some aspects, in the amino acid sequence of SEQ ID NO: 48, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In some aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 48, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:26, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:27, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:28.

[0412] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35 to 47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52, including post-translational modifications of those sequences. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 47 and a VL sequence of SEQ ID NO: 48, including post-translational modifications of those sequences.

[0413] In one aspect, the VL sequence comprises a V4M mutation, a P43A mutation, a F46L mutation, a C90Q mutation, or a combination thereof (e.g., according to Kabat numbering). In one aspect, the VH comprises a G49S mutation, a K71R mutation, a S73N mutation, or a combination thereof (e.g., according to Kabat numbering).

[0414] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59. In one aspect, the antibody comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: (a) an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59; and (b) a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54.

[0415] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 31, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 32; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 26, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 27, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 28. In one aspect, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 47 and a VL sequence of SEQ ID NO: 48.

[0416] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:55 and the light chain of SEQ ID NO:56.

[0417] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:60 and the light chain of SEQ ID NO:56.

[0418] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the heavy chain of the antibody comprises a shortened C-terminus, wherein one or two of the C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending in PG. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 111 and a light chain of SEQ ID NO: 56. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 113 and a light chain of SEQ ID NO: 56.

[0419] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody does not bind to a CCR8 ligand. In one aspect, the anti-CCR8 antibody does not have CCR8 ligand blocking activity. In one aspect, the anti-CCR8 antibody is a full-length antibody. In one aspect, the CCR8 ligand is CCL1.

[0420] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody binds to CCR8 independently of tyrosine sulfation of CCR8 (ie, is sulfation-independent).

[0421] In another aspect of any one of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody is a defucosylated antibody variant. In one aspect, the defucosylated antibody variant has enhanced FcγRIIIa receptor binding. In one aspect, the defucosylated anti-CCR8 antibody variant has enhanced antibody-dependent cellular cytotoxicity (ADCC). In one aspect, the anti-CCR8 defucosylated antibody variant has antibody-dependent cellular phagocytosis (ADCP) activity.

[0422] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody has improved antibody stability. In one aspect, the anti-CCR8 antibody has low aggregation, good solubility and / or low viscosity. In certain aspects of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody has about 1×10 -12 M is about 1×10 -11 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 5×10 -12 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 4×10 -12 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 3×10 -12 M of K D .

[0423] In one aspect, the anti-CCR8 antibody is designated in the present disclosure as "hu.Ab5.H13L1", which may be fucosylated or defucosylated, which optionally contains one or more heavy chain mutations at G236A and I331E, and which optionally comprises a shortened C-terminus of the heavy chain, wherein one or two of the C-terminal amino acid residues have been removed. In some embodiments, the heavy chain mutations are numbered according to the EU index.

[0424] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including chimeric, humanized or human antibodies. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0425] (ii) Embodiments of Ab4 and fragments thereof

[0426] In one aspect, the present disclosure provides an anti-CCR8 antibody comprising at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO:6; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO:7; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO:1; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO:2; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO:3. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to both human CCR8 and cynomolgus monkey CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0427] In one aspect, the present disclosure provides an antibody comprising at least one, at least two, or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO:6; and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO:7. In one aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO:7. In another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO:7; and CDR-L3 comprising an amino acid sequence of SEQ ID NO:3. In yet another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO:7; CDR-L3 comprising an amino acid sequence of SEQ ID NO:3; and CDR-H2 comprising an amino acid sequence of SEQ ID NO:6. In yet another aspect, the antibody comprises: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:6; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:7.

[0428] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 1; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 2; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 3. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 1; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 2; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 3.

[0429] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:6; and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:7; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:1; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:2; and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:3. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to both human CCR8 and cynomolgus monkey CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0430] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:1, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:2, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:3.

[0431] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequences selected from the group consisting of SEQ ID NOs: 10 to 21. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequences selected from the group consisting of SEQ ID NOs: 22 to 25. In another aspect, the anti-CCR8 antibody comprises a CDR sequence of the VH sequence selected from the group consisting of SEQ ID NOs: 10 to 21 and a CDR sequence of the VL sequence selected from the group consisting of SEQ ID NOs: 22 to 25.

[0432] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 21. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 24. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 21. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 24.

[0433] In another aspect, the anti-CCR8 antibody comprises CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain selected from the group consisting of SEQ ID NOs: 10 to 21 and CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain selected from the group consisting of SEQ ID NOs: 22 to 25.

[0434] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:21 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:24.

[0435] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21 and a framework having at least 95% sequence identity with a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21. In another aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21 and a framework having at least 98% sequence identity with a framework amino acid sequence of a VH domain selected from the group consisting of SEQ ID NOs: 10 to 21.

[0436] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 21 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 21. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 21 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 21. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 21 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 21. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 21; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 21.

[0437] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25 and a framework having at least 95% sequence identity with a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25. In another aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25 and a framework having (especially at least) at least 98% sequence identity with a framework amino acid sequence of a VL domain selected from the group consisting of SEQ ID NOs: 22 to 25.

[0438] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 24 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 24. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 24 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 24. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 24 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 24. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 24; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 24.

[0439] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:6; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:1; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:3, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs:10 to 21; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs:10 to 21; The amino acid sequences of the group consisting of SEQ ID NOs: 22 to 25 have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with an antibody comprising a VH sequence selected from the group consisting of SEQ ID NOs: 10 to 21 and a VL sequence selected from the group consisting of SEQ ID NOs: 22 to 25 (K D ) is up to 10 times lower or up to 10 times higher.

[0440] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:6; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:1; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:3, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO:21; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO:21; NO:24 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:21. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:24. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 21 and the VL sequence of SEQ ID NO: 24 (K D ) is up to 10 times lower or up to 10 times higher.

[0441] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21. In one aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21. In certain aspects, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In some aspects, in the amino acid sequence selected from the group consisting of SEQ ID NO: 10 to 21, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In some aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises a VH sequence selected from the group consisting of SEQ ID NO: 10 to 21, including post-translational modifications of the sequence. In a specific aspect, VH comprises one, two or three CDRs selected from the following: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5, (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 6, and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 7. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25. In one aspect, the anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25. In certain aspects, the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In some aspects, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25. In some aspects, the substitution, insertion or deletion occurs in a region outside of the CDR (i.e., in the FR).Optionally, the anti-CCR8 antibody comprises a VL sequence selected from the group consisting of SEQ ID NOs: 22 to 25, including post-translational modifications of the sequence. In a specific aspect, the VL comprises one, two or three CDRs selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 1, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 2, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 3.

[0442] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 21. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 21. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 21, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 21, including post-translational modifications of the sequence. In a specific aspect, VH comprises one, two or three CDRs selected from the following: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5, (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 6, (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 7. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 24. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 24. In certain aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 24, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 24, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from the following: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:1, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:2, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:3.

[0443] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence selected from the group consisting of SEQ ID NOs: 10 to 21 and a VL sequence selected from the group consisting of SEQ ID NOs: 22 to 25, including post-translational modifications of those sequences. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 21 and a VL sequence of SEQ ID NO: 24, including post-translational modifications of those sequences.

[0444] In one aspect, the VL sequence comprises a Y2I mutation. In another aspect, the VH sequence comprises a S73N mutation, a V78L mutation, a T76N mutation, a F91Y mutation, and a P105Q mutation, or a combination thereof (e.g., according to Kabat numbering).

[0445] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59. In one aspect, the antibody comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: (a) an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59; and (b) a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54.

[0446] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 6, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 7; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 1, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 2, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 3. In one aspect, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 21 and a VL sequence of SEQ ID NO: 24.

[0447] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:57 and the light chain of SEQ ID NO:58.

[0448] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:61 and the light chain of SEQ ID NO:58.

[0449] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the heavy chain of the antibody comprises a shortened C-terminus, wherein one or two of the C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending in PG. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 112 and a light chain of SEQ ID NO: 58. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 114 and a light chain of SEQ ID NO: 58.

[0450] In another aspect of any one of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody binds to a CCR8 ligand. In one aspect, the anti-CCR8 antibody has an antagonistic effect against a CCR8 ligand. In one aspect, the anti-CCR8 antibody has CCR8 ligand blocking activity. In one aspect, the anti-CCR8 antibody is a neutralizing antibody. In one aspect, the CCR8 ligand is CCL1.

[0451] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody binds to CCR8 independently of tyrosine sulfation of CCR8 (ie, is sulfation-independent).

[0452] In another aspect of any one of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody is a defucosylated antibody variant. In one aspect, the defucosylated antibody variant has enhanced FcγRIIIa receptor binding. In one aspect, the defucosylated anti-CCR8 antibody variant has enhanced antibody-dependent cellular cytotoxicity (ADCC). In one aspect, the anti-CCR8 defucosylated antibody variant has antibody-dependent cellular phagocytosis (ADCP) activity.

[0453] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody has improved antibody stability. In one aspect, the anti-CCR8 antibody has low aggregation, good solubility and / or low viscosity. In certain aspects of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the antibody has about 1×10 -11 M is about 5×10 -11 M of K D In certain aspects, the antibody that binds to CCR8 has a binding affinity of about 2×10 -11 M of K D .

[0454] In one aspect, the anti-CCR8 antibody is designated in the present disclosure as "hu.Ab4.H12L3", which can be fucosylated or defucosylated, which optionally contains one or more heavy chain mutations at G236A and I331E, and which optionally comprises a shortened C-terminus of the heavy chain, wherein one or two of the C-terminal amino acid residues have been removed. In some cases, the heavy chain mutations are numbered according to the EU index.

[0455] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including chimeric, humanized or human antibodies. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0456] (iii) Embodiments of Ab1 and its fragments

[0457] In one aspect, the present disclosure provides an anti-CCR8 antibody comprising at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 73; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 74; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 75. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0458] In one aspect, the present disclosure provides an antibody comprising at least one, at least two, or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84; and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85. In one aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85. In another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85; and CDR-L3 comprising an amino acid sequence of SEQ ID NO: 75. In yet another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85; CDR-L3 comprising an amino acid sequence of SEQ ID NO: 75; and CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84. In yet another aspect, the antibody comprises: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:82 or SEQ ID NO:83; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:84; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:85.

[0459] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 75. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 75.

[0460] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83; (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84; and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 73; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 74; and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 75. In certain aspects, the anti-CCR8 antibody comprises all six of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0461] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:82 or SEQ ID NO:83, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:84, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:85; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:73, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:74, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:75.

[0462] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 95. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 94. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 95. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 94.

[0463] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:95 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:94.

[0464] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 95 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 95. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 95 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 95. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 95 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 95. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 95; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 95.

[0465] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 94 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 94. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 94 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 94. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 94 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 94. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 94; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 94.

[0466] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 84; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 85; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 75, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 95; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 95; NO:94 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:95. In one aspect, the VL domain has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:94. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 95 and the VL sequence of SEQ ID NO: 94 (K D ) is up to 10 times lower or up to 10 times higher.

[0467] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 95. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 95. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 95, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 95, including post-translational modifications of the sequence. In a specific aspect, VH comprises one, two or three CDRs selected from the following: (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 94. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 94. In certain aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 94, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 94, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:73, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:74, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:75.

[0468] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 95 and a VL sequence of SEQ ID NO: 94, including post-translational modifications of those sequences.

[0469] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59. In one aspect, the antibody comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: (a) an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59; and (b) a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54.

[0470] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 84, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 85; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 73, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 74, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 75. In one aspect, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 95 and a VL sequence of SEQ ID NO: 94.

[0471] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:101 and the light chain of SEQ ID NO:100.

[0472] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the heavy chain of the antibody comprises a shortened C-terminus, wherein one or two of the C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending in PG. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 115 and a light chain of SEQ ID NO: 100.

[0473] In one aspect, an anti-CCR8 antibody is designated in the present disclosure as "hu.Abl.H1L1", which may be fucosylated or defucosylated, which optionally contains one or more heavy chain mutations at G236A and I331E, and which optionally comprises a shortened C-terminus of the heavy chain, wherein one or two of the C-terminal amino acid residues have been removed. In some cases, the heavy chain mutations are numbered according to the EU index.

[0474] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including chimeric, humanized or human antibodies. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0475] (iv) Embodiments of Ab2 and fragments thereof

[0476] In one aspect, the present disclosure provides an anti-CCR8 antibody comprising at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 88; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 89; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 76; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 77; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 78. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0477] In one aspect, the present disclosure provides an antibody comprising at least one, at least two, or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89. In one aspect, the antibody comprises: CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89. In another aspect, the antibody comprises: CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; and CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78. In yet another aspect, the antibody comprises: CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78; and CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88. In yet another aspect, the antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:86 or SEQ ID NO:87; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:88; and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:89.

[0478] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78.

[0479] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87; (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 88; and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 89; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 76; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 77; and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 78. In certain aspects, the anti-CCR8 antibody comprises all six of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0480] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 89; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 76, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 77, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 78.

[0481] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 97. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 96. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 97. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 96.

[0482] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:97 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:96.

[0483] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 97 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 97. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 97 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 97. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 97 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 97. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 97; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 97.

[0484] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 96 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 96. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 96 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 96. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 96 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 96. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 96; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 96.

[0485] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 97; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 97; NO:96 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:97. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:96. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 97 and the VL sequence of SEQ ID NO: 96 (K D ) is up to 10 times lower or up to 10 times higher.

[0486] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 97. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 97. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 97, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 97, including post-translational modifications of the sequence. In a specific aspect, the VH comprises one, two or three CDRs selected from the following: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 96. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 96. In certain aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 96, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 96, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:76, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:75, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:78.

[0487] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 97 and a VL sequence of SEQ ID NO: 96, including post-translational modifications of those sequences.

[0488] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59. In one aspect, the antibody comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: (a) an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59; and (b) a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54.

[0489] On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 78.

[0490] In one aspect, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO:97 and the VL sequence of SEQ ID NO:96.

[0491] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:103 and the light chain of SEQ ID NO:102.

[0492] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the heavy chain of the antibody comprises a shortened C-terminus, wherein one or two of the C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending in PG. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 116 and a light chain of SEQ ID NO: 102.

[0493] In one aspect, the anti-CCR8 antibody is designated in the present disclosure as "hu.Ab2.H1L1", which can be fucosylated or defucosylated, which optionally contains one or more heavy chain mutations at G236A and I331E, and which optionally comprises a shortened C-terminus of the heavy chain, wherein one or two of the C-terminal amino acid residues have been removed. In some cases, the heavy chain mutations are numbered according to the EU index.

[0494] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including chimeric, humanized or human antibodies. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0495] (v) Embodiments of Ab3 and fragments thereof

[0496] In one aspect, the present disclosure provides an anti-CCR8 antibody comprising at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 92; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 79; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 80; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 81. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0497] In one aspect, the present disclosure provides an antibody comprising at least one, at least two, or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 92; and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93. In one aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93. In another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93; and CDR-L3 comprising an amino acid sequence of SEQ ID NO: 81. In yet another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93; CDR-L3 comprising an amino acid sequence of SEQ ID NO: 81; and CDR-H2 comprising an amino acid sequence of SEQ ID NO: 92. In yet another aspect, the antibody comprises: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:90 or SEQ ID NO:91; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:92; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:93.

[0498] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 79; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 80; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 81. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 79; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 80; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 81.

[0499] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91; (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 92; and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 79; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 80; and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 81. In certain aspects, the anti-CCR8 antibody comprises all six of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a human antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a humanized antibody. In some aspects, the anti-CCR8 antibody is a full-length antibody that binds to human CCR8 and is a chimeric antibody.

[0500] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO:90 or SEQ ID NO:91, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO:92, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO:93; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO:79, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO:80, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO:81.

[0501] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 99. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 98. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 99. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 98.

[0502] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:99 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:98.

[0503] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 99 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 99. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 99 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 99. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 99 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 99. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 99; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 99.

[0504] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 98 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 98. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 98 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 98. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 98 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 98. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 98; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 98.

[0505] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 92; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 93; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 79; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 80; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 81, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 99; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 99; NO:98 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:99. In one aspect, the VL domain has at least 95% sequence identity with the amino acid sequence of SEQ ID NO:98. In one aspect, the antibody binds to CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 99 and the VL sequence of SEQ ID NO: 98 (K D ) is up to 10 times lower or up to 10 times higher.

[0506] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 99. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 99. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 99, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside of the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 99, including post-translational modifications of the sequence. In a specific aspect, the VH comprises one, two or three CDRs selected from the following: SEQ ID NO: 90 or SEQ ID NO: 91, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 92, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 93. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 98. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 98. In certain aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 98, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 98, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:79, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:80, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:81.

[0507] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 99 and a VL sequence of SEQ ID NO: 98, including post-translational modifications of those sequences.

[0508] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59. In one aspect, the antibody comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54. In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: (a) an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO: 59; and (b) a kappa constant domain comprising the amino acid sequence of SEQ ID NO: 54.

[0509] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 92, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 93; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 79, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 80, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 81. In one aspect, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 99 and a VL sequence of SEQ ID NO: 98.

[0510] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:105 and the light chain of SEQ ID NO:104.

[0511] In another aspect of any of the above embodiments, an anti-CCR8 antibody is provided, wherein the heavy chain of the antibody comprises a shortened C-terminus, wherein one or two of the C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending in PG. In one aspect, the anti-CCR8 antibody comprises a heavy chain of SEQ ID NO: 117 and a light chain of SEQ ID NO: 104.

[0512] In one aspect, the anti-CCR8 antibody is designated in the present disclosure as "hu.Ab3.H1L1", which can be fucosylated or defucosylated, which optionally contains one or more heavy chain mutations at G236A and I331E, and which optionally comprises a shortened C-terminus of the heavy chain, wherein one or two of the C-terminal amino acid residues have been removed. In some cases, the heavy chain mutations are numbered according to the EU index.

[0513] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including chimeric, humanized or human antibodies. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0514] (vi) Embodiment of mouse substitute

[0515] In one aspect, the present disclosure provides an anti-CCR8 antibody that binds to mouse CCR8 and comprises at least one, at least two, at least three, at least four, at least five or at least six CDRs selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 67; (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68; (d) CDR-L1 comprising an amino acid sequence of SEQ ID NO: 62; (e) CDR-L2 comprising an amino acid sequence of SEQ ID NO: 63; and (f) CDR-L3 comprising an amino acid sequence of SEQ ID NO: 64. In certain aspects, the anti-CCR8 antibody comprises all six CDRs of the above-mentioned CDRs. In certain aspects, the anti-CCR8 antibody is a full-length antibody. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to mouse CCR8. In certain aspects, the anti-CCR8 antibody is a full-length antibody that binds to mouse CCR8 and is a chimeric antibody (eg, a rabbit and mouse chimera).

[0516] In one aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (a) CDR-H1 comprising an amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66; (b) CDR-H2 comprising an amino acid sequence of SEQ ID NO: 67; and (c) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68. In one aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68. In another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68; and CDR-L3 comprising an amino acid sequence of SEQ ID NO: 64. In yet another aspect, the antibody comprises: CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68; CDR-L3 comprising an amino acid sequence of SEQ ID NO: 64; and CDR-H2 comprising an amino acid sequence of SEQ ID NO: 6. In yet another aspect, the antibody comprises: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:65 or SEQ ID NO:66; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO:67; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:68.

[0517] In another aspect, the present disclosure provides an antibody comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 62; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 63; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 64. In one aspect, the antibody comprises (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 62; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 63; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 64.

[0518] In another aspect, an antibody as described herein comprises: (a) a VH domain comprising at least one, at least two or all three VH CDR sequences selected from the group consisting of: (i) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66; (ii) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 67; and (iii) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68; and (b) a VL domain comprising at least one, at least two or all three VL CDR sequences selected from the group consisting of: (i) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 62; (ii) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 63; and (iii) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 64.

[0519] On the other hand, the present disclosure provides an antibody comprising: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:65 or SEQ ID NO:66, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:67, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:68; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:62, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:63, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:64.

[0520] In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 70. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 69. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VH sequence of SEQ ID NO: 70. In another aspect, the anti-CCR8 antibody comprises one or more of the CDR sequences of the VL sequence of SEQ ID NO: 69.

[0521] In another aspect, an anti-CCR8 antibody comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH domain of SEQ ID NO:70 and the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL domain of SEQ ID NO:69.

[0522] In one aspect, the anti-CCR8 antibody comprises one or more heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 70 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 70. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 70 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VH domain of SEQ ID NO: 70. In one aspect, the anti-CCR8 antibody comprises three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 70 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 70. In another aspect, the anti-CCR8 antibody comprises: three heavy chain CDR amino acid sequences of the VH domain of SEQ ID NO: 70; and a framework having at least 98% sequence identity with the framework amino acid sequence of the VH domain of SEQ ID NO: 70.

[0523] In one aspect, the anti-CCR8 antibody comprises one or more light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 69 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 69. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 69 and a framework having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the framework amino acid sequence of the VL domain of SEQ ID NO: 69. In one aspect, the anti-CCR8 antibody comprises three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 69 and a framework having at least 95% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 69. In another aspect, the anti-CCR8 antibody comprises: three light chain CDR amino acid sequences of the VL domain of SEQ ID NO: 69; and a framework having (especially at least) at least 98% sequence identity with the framework amino acid sequence of the VL domain of SEQ ID NO: 69.

[0524] In one aspect, the anti-CCR8 antibody comprises: (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 62; (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 63; and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 64, and a VH domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 70; and a VL domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 70; NO:69 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In one aspect, the VH domain has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:70. In one aspect, the VL domain has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:69. In one aspect, the antibody binds to mouse CCR8 with a dissociation constant (K D ) with the antibody comprising the VH sequence of SEQ ID NO: 70 and the VL sequence of SEQ ID NO: 69 (K D ) is up to 10 times lower or up to 10 times higher.

[0525] In another aspect, the anti-CCR8 antibody comprises: a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 70. In one aspect, the anti-CCR8 antibody comprises a heavy chain variable domain (VH) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 70. In certain aspects, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind to mouse CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 70, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, the substitution, insertion or deletion occurs in a region outside the CDR (i.e., in the FR). Optionally, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 70, including post-translational modifications of the sequence. In a specific aspect, the VH comprises one, two or three CDRs selected from the following: SEQ ID NO: 65 or SEQ ID NO: 66, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68. On the other hand, an anti-CCR8 antibody is provided, wherein the antibody comprises: a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 69. In one aspect, an anti-CCR8 antibody comprises: a light chain variable domain (VL) sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 69. In certain aspects, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to a reference sequence, but an anti-CCR8 antibody comprising the sequence retains the ability to bind to CCR8. In certain aspects, in the amino acid sequence of SEQ ID NO: 69, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted. In certain aspects, substitutions, insertions or deletions occur in regions outside of CDR (i.e., in FR). Optionally, an anti-CCR8 antibody comprises a VL sequence of SEQ ID NO: 69, including post-translational modifications of the sequence.In a specific aspect, VL comprises one, two or three CDRs selected from: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO:62, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO:63, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO:64.

[0526] In another aspect, an anti-CCR8 antibody is provided, wherein the antibody comprises a VH sequence as in any aspect provided above and a VL sequence as in any aspect provided above. In one aspect, the antibody comprises a VH sequence of SEQ ID NO: 70 and a VL sequence of SEQ ID NO: 69, including post-translational modifications of those sequences.

[0527] In another aspect, an anti-CCR8 antibody that binds to mouse CCR8 is provided, wherein the antibody comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising an amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66, (b) a CDR-H2 comprising an amino acid sequence of SEQ ID NO: 67, and (c) a CDR-H3 comprising an amino acid sequence of SEQ ID NO: 68; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising an amino acid sequence of SEQ ID NO: 62, (e) a CDR-L2 comprising an amino acid sequence of SEQ ID NO: 63, and (f) a CDR-L3 comprising an amino acid sequence of SEQ ID NO: 64. In one aspect, the anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 70 and a VL sequence of SEQ ID NO: 69.

[0528] In one aspect, the anti-CCR8 antibody comprises the heavy chain of SEQ ID NO:72 and the light chain of SEQ ID NO:71.

[0529] In a further aspect, the anti-CCR8 antibody according to any one of the above aspects is a monoclonal antibody, including a chimeric antibody. In one aspect, the anti-CCR8 antibody is an antibody fragment, such as Fv, Fab, Fab', scFv, diabody or F(ab'). 2 Fragment.

[0530] (vii) Other Implementation Schemes

[0531] In a further aspect, the anti-CCR8 antibody according to any of the above aspects may include any of the features described in Sections 1 to 5 below, alone or in combination:

[0532] 1. Antibody fragments

[0533] In certain aspects, the antibodies provided herein are antibody fragments.

[0534] In one aspect, the antibody fragment is Fab', Fab'-SH or F(ab') 2 Fragments, particularly Fab fragments. Papain digestion of intact antibodies produces two identical antigen-binding fragments called "Fab" fragments, each of which contains the heavy chain variable domain and the light chain variable domain (VH and VL, respectively) and the constant domain of the light chain (CL) and the first constant domain of the heavy chain (CH1). Therefore, the term "Fab fragment" refers to an antibody fragment that includes a light chain comprising the VL domain and the CL domain, and a heavy chain fragment comprising the VH domain and the CH1 domain. Fab' fragments differ from Fab fragments in that Fab' fragments have added residues at the carboxyl terminus of the CH1 domain, which include one or more cysteines from the antibody hinge region. Fab'-SH is a Fab' fragment in which the cysteine ​​residues of the constant domains have a free thiol group. Pepsin treatment produces F(ab') 2 A fragment having two antigen binding sites (two Fab fragments) and a portion of the Fc region. Regarding Fab and F(ab') containing salvage receptor binding epitope residues and having increased in vivo half-life 2 For a discussion of fragments, see U.S. Patent No. 5,869,046.

[0535] In another aspect, the antibody fragment is a diabody, a triabody, or a tetrabody. A "diabody" is an antibody fragment with two antigen binding sites that can be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0536] In another aspect, the antibody fragment is a single-chain Fab fragment. "Single-chain Fab fragment" or "scFab" is a polypeptide consisting of an antibody heavy chain variable domain (VH), an antibody heavy chain constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL) and a linker, wherein the antibody domain and the linker have one of the following orders in the N-terminal to C-terminal direction: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CL-linker-VL-CH1, or d) VL-CH1-linker-VH-CL. In particular, the linker is a polypeptide of at least 30 amino acids, preferably between 32 and 50 amino acids. The single-chain Fab fragment is stabilized via a natural disulfide bond between the CL domain and the CH1 domain. Furthermore, these single-chain Fab fragments can be further stabilized by generating interchain disulfide bonds via the insertion of cysteine ​​residues (eg, position 44 in the variable heavy chain and position 100 in the variable light chain according to Kabat numbering).

[0537] On the other hand, the antibody fragment is a single-chain variable fragment (scFv). "Single-chain variable fragment" or "scFv" is a fusion protein of the heavy chain variable domain (VH) and the light chain variable domain (VL) of an antibody, which is connected by a linker. In particular, the linker is a short polypeptide of 10 to about 25 amino acids, and is generally rich in glycine to obtain flexibility, and rich in serine or threonine to obtain solubility, and the N-terminus of VH can be connected to the C-terminus of VL, or vice versa. Although the constant region is removed and the linker is introduced, the protein still retains the specificity of the original antibody. For a review of scFv fragments, see, for example, Plückthun in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, ed., (Springer-Verlag, New York), described in pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,587,458.

[0538] On the other hand, the antibody fragment is a single domain antibody. A "single domain antibody" is an antibody fragment that contains all or part of the heavy chain variable domain of an antibody or all or part of the light chain variable domain of an antibody. In certain aspects, a single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516B1).

[0539] Antibody fragments can be prepared by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and recombinant production by recombinant host cells (eg, E. coli), as described herein.

[0540] 2. Chimeric and humanized antibodies

[0541] In certain aspects, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described in, for example, U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81: 6851-6855 (1984). In one example, a chimeric antibody includes a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In another example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0542] In some aspects, chimeric antibodies are humanized antibodies. Generally, non-human antibodies are humanized to reduce immunogenicity to people, while retaining the specificity and affinity of the parent non-human antibody. Generally, humanized antibodies comprise one or more variable domains, wherein CDR (or its part) is derived from non-human antibodies, and FR (or its part) is derived from human antibody sequences. Humanized antibodies will optionally also comprise at least a portion of human constant regions. In some embodiments, some FR residues in humanized antibodies are replaced by corresponding residues from non-human antibodies (for example, antibodies from which CDR residues are derived), for example, to restore or improve antibody specificity or affinity.

[0543] Humanized antibodies and methods for their preparation are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, for example, in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method for FR shuffling).

[0544] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best fit" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)). )); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272: 10678-10684 (1997); and Rosok et al., J. Biol. Chem. 271: 22611-22618 (1996)).

[0545] 3. Human Antibodies

[0546] In certain aspects, the antibodies provided herein are human antibodies. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[0547] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce complete human antibodies or complete antibodies with human variable regions in response to antigenic stimulation. Such animals typically contain all or part of a human immunoglobulin locus that replaces an endogenous immunoglobulin locus, or exists outside the chromosome of the animal or is randomly integrated into the chromosome of the animal. In such transgenic mice, the endogenous immunoglobulin locus is typically inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23: 1117-1125 (2005). See also, for example, description of XENOMOUSE TM US Patent Nos. 6,075,181 and 6,150,584 for the technology; description US Patent No. 5,770,429 for technology; describes KM US Patent No. 7,041,870, and describes The human variable regions from intact antibodies produced by such animals can be further modified, for example by combining with different human constant regions.

[0548] Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human hybrid myeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Human antibodies produced via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Additional methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26 (4): 265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20 (3): 927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27 (3): 185-91 (2005).

[0549] Human antibodies can also be produced by isolating variable domain sequences selected from human phage display libraries. Such variable domain sequences can then be combined with expected human constant domains. The technology for selecting human antibodies from antibody libraries is described below.

[0550] 4. Multispecific Antibodies

[0551] In some aspects, the antibody provided herein is a multispecific antibody, for example, a bispecific antibody." Multispecific antibody " is a monoclonal antibody with binding specificity to at least two different sites (i.e., different epitopes on different antigens or different epitopes on the same antigen). In some aspects, a multispecific antibody has three or more binding specificities. In some aspects, one of the binding specificities is for CCR8, and another specificity is for any other antigen. In some aspects, a bispecific antibody can be combined with two (or more) different epitopes of CCR8. Multispecific (e.g., bispecific) antibodies can also be used to localize cytotoxic agents or cells to cells expressing CCR8. Multispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0552] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, Nature, 305:537 (1983)) and “knob-in-hole” engineering (see, e.g., U.S. Pat. No. 5,731,168, and Atwell et al., J. Mol. Biol. 270:26 (1997)). Multispecific antibodies can also be prepared by engineering electrostatic manipulation effects for preparing antibody Fc-heterodimer molecules (see, e.g., WO 2009 / 089004); cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980, and Brennan et al., Science, 229:81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5):1547-1553 (1992) and WO 2011 / 034605); using common light chain technology to avoid light chain mispairing problems (see, e.g., WO 98 / 50431); using "diabody" technology for preparing bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and preparing trispecific antibodies as described, e.g., in Tutt et al., J. Immunol. 147:60 (1991).

[0553] 5. Antibody variants

[0554] In some aspects, it is contemplated that the amino acid sequence variants of the antibodies provided herein. For example, it may be desirable to change the binding affinity and / or other biological properties of the antibody. The amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, the deletion of residues in the antibody amino acid sequence, and / or insertion and / or substitution. Any combination of deletion, insertion and substitution can be performed to achieve the final construct, provided that the final construct has a desired feature, for example, antigen binding.

[0555] a) Substitution, insertion and deletion variants

[0556] In certain aspects, antibody variants with one or more amino acid substitutions are provided. Sites of interest for substitution mutagenesis include CDRs and FRs.

[0557] In one aspect, the VL sequence of an antibody disclosed herein comprises a V4M mutation, a P43A mutation, a F46L mutation, a C90Q mutation, or a combination thereof. In one aspect, the VH of an antibody disclosed herein comprises a G49S mutation, a K71R mutation, a S73N mutation, or a combination thereof. In one aspect, the VL sequence of an antibody disclosed herein comprises a Y2I mutation. In one aspect, the VH sequence of an antibody disclosed herein comprises a S73N mutation, a V78L mutation, a T76N mutation, a F91Y mutation, and a P105Q mutation, or a combination thereof. In some cases, any of the above mutations are according to the Kabat method.

[0558]

[0559] Row number.

[0560] Conservative substitutions are shown under the heading "Conservative Substitutions" in Table 2. More substantial changes are provided under the heading "Exemplary Substitutions" in Table 2 and are further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into an antibody of interest and the products screened for a desired activity (e.g., maintained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC).

[0561] Amino acids can be grouped according to common side chain properties:

[0562] (1) Hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;

[0563] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0564] (3) Acidic: Asp, Glu;

[0565] (4) Basic: His, Lys, Arg;

[0566] (5) Residues that affect chain orientation: Gly, Pro;

[0567] (6) Aromatic: Trp, Tyr, Phe.

[0568] Non-conservative substitutions will entail exchanging a member of one of these classes for a member of another class.

[0569] One type of substitution variant involves replacing one or more hypervariable region residues of a parent antibody (e.g., a humanized antibody or a human antibody). Typically, one or more resulting variants selected for further study will have modifications (e.g., improvements) and / or will substantially retain certain biological properties of the parent antibody in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parent antibody. Exemplary substitution variants are affinity matured antibodies, which can be conveniently generated, for example, using affinity maturation techniques based on phage display such as those described herein. Briefly, one or more CDR residues are mutated and the variant antibodies are displayed on phage and screened for specific biological activity (e.g., binding affinity).

[0570] For example, changes (e.g., substitutions) can be made in the CDR to improve antibody affinity. Such changes can occur in CDR "hot spots", i.e., residues encoded by codons that undergo high-frequency mutations during somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207: 179-196 (2008)) and / or residues in contact with the antigen (detecting the binding affinity of the resulting variant VH or VL). Affinity maturation achieved by construction and reselection from a secondary library has been described, for example, by Hoogenboom et al. in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)). In certain aspects of affinity maturation, diversity is introduced into variable genes selected for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is subsequently screened to identify any antibody variants with the desired affinity. Another method of introducing diversity involves a CDR-directed approach in which several CDR residues (e.g., 4 to 6 residues at a time) are randomized. The CDR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. In particular, CDR-H3 and CDR-L3 are often targeted.

[0571] In some aspects, substitutions, insertions or deletions may occur within one or more CDRs, as long as such changes do not substantially reduce the ability of the antibody to bind to antigen. For example, conservative changes (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in the CDRs. Such changes may be, for example, outside of the antigen contact residues in the CDRs. In some variant VH and VL sequences provided above, each CDR either remains unchanged or contains no more than one, two or three amino acid substitutions.

[0572] The method that can be used to identify antibody residues or regions that can be targeted for mutagenesis is called "alanine scanning mutagenesis", as described in Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, a residue or a group of target residues (e.g., charged residues such as Arg, Asp, His, Lys and Glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with the antigen is affected. Other substitutions can be introduced at amino acid positions that exhibit functional sensitivity to the initial substitution. Alternatively or additionally, the contact points between the antibody and the antigen can be identified using the crystal structure of the antigen-antibody complex. Such contact residues and adjacent residues that are candidates for substitution can be targeted or eliminated. Variants can be screened to determine whether they have the desired properties.

[0573] Amino acid sequence insertions include amino and / or carboxyl terminal fusions ranging in length from one residue to a polypeptide containing one hundred or more residues, and intrasequence insertions of one or more amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertion variants of antibody molecules include fusing the N-terminus or C-terminus of the antibody to an enzyme (e.g., for ADEPT (antibody directed enzyme prodrug therapy)) or a polypeptide that increases the serum half-life of the antibody.

[0574] b) Glycosylation variants

[0575] In certain aspects, the antibodies provided herein are altered to increase or decrease the extent of antibody glycosylation. Glycosylation sites can be conveniently added or deleted to an antibody by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0576] When antibody comprises Fc district, the oligosaccharide connected thereto can be changed.Native antibodies produced by mammalian cells usually comprise the biantennary oligosaccharide of side chain, and this biantennary oligosaccharide is usually connected to the Asn297 of CH2 domain in Fc district by N-bonding.See, for example, Wright et al. TIBTECH 15:26-32 (1997).Oligosaccharide can include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid, and the fucose of GlcNAc in the " backbone " of biantennary oligosaccharide structure.In some aspects, the oligosaccharide in antibody as described herein can be modified, so as to produce antibody variants with some improved characteristics.

[0577] In one aspect, antibody variants having non-fucosylated oligosaccharides, i.e., oligosaccharide structures lacking fucose attached (directly or indirectly) to the Fc region are provided. Such non-fucosylated oligosaccharides (also referred to as "defucosylated" oligosaccharides) are particularly N-linked oligosaccharides that lack a fucose residue attached to the first GlcNAc in the stem of the biantennary oligosaccharide structure, and such antibodies are further referred to herein as "defucosylated antibodies". In one aspect, antibody variants having an increased proportion of non-fucosylated oligosaccharides in the Fc region compared to a natural or parent antibody are provided. For example, the proportion of non-fucosylated oligosaccharides can be at least about 20%, at least about 40%, at least about 60%, at least about 80%, or even about 100% (i.e., there is no fucosylated oligosaccharide). In certain embodiments, the defucosylation ratio is about 65% to about 100%, about 80% to about 100%, or about 80% to about 95%. The percentage of non-fucosylated oligosaccharides, as described, for example, in WO 2006 / 082515, as measured by MALDI-TOF mass spectrometry, is the (average) amount of oligosaccharides lacking a fucose residue relative to the sum of all oligosaccharides (e.g., complex, hybrid and high mannose structures) linked to Asn 297. Asn297 refers to the asparagine residue located at approximately position 297 in the Fc region (EU numbering of residues in the Fc region); however, due to minor sequence variations in antibodies, Asn297 may also be located about ±3 amino acids upstream or downstream of position 297, i.e., between position 294 and position 300, e.g., Asn 299. Such antibodies having an increased proportion of non-fucosylated oligosaccharides in the Fc region may have improved FcγRIIIa receptor binding and / or improved effector function, in particular improved ADCC function. See, for example, US2003 / 0157108 and US2004 / 0093621.

[0578] In one aspect, the present disclosure provides a defucosylated antibody variant with enhanced FcγRIIIa receptor binding. In one aspect, the present disclosure provides a defucosylated antibody variant with enhanced antibody-dependent cellular cytotoxicity (ADCC). In one aspect, the present disclosure provides a defucosylated antibody variant with antibody-dependent cellular phagocytosis (ADCP) activity.

[0579] Examples of cell lines capable of producing antibodies with reduced fucosylation include Lec13 CHO cells lacking protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US 2003 / 0157108; and WO 2004 / 056312, especially in Example 11), and knockout cell lines, such as α-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614-622 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4): 680-688 (2006); and WO2003 / 085107), or cells with reduced or abolished GDP-fucose synthesis or transporter activity (see, e.g., US2004259150, US2005031613, US2004132140, US2004110282). See also Pereira et al., MABS (2018) 693-711.

[0580] In a further aspect, the antibody variant provides a bisected oligosaccharide, for example, wherein the biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. As described above, such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described in, for example, Umana et al., Nat Biotechnol 17, 176-180 (1999); Ferrara et al., Biotechn Bioeng 93, 851-861 (2006); WO 99 / 54342; WO 2004 / 065540, WO 2003 / 011878.

[0581] Also provided are antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region. Such antibody variants may have improved CDC function. Such antibody variants are described in, for example, WO 1997 / 30087, WO 1998 / 58964, and WO 1999 / 22764.

[0582] c) Fc region variants

[0583] In certain aspects, one or more amino acid modifications can be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant can comprise a human Fc region sequence (e.g., human IgG 1 IgG 2 IgG 3 or IgG 4 Fc region) comprising an amino acid modification (e.g., substitution) at one or more amino acid positions.

[0584] In certain aspects, the present invention contemplates antibody variants with some but not all effector functions, making them ideal candidates for applications where the in vivo half-life of the antibody is important and certain effector functions (e.g., complement dependent cytotoxicity (CDC) and antibody dependent cell-mediated cytotoxicity (ADCC)) are unnecessary or harmful. In vitro and / or in vivo cytotoxicity assays can be performed to confirm the reduction / depletion of CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcγR binding (and therefore may lack ADCC activity), but retains FcRn binding ability. NK cells, the main cells that mediate ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9: 457-492 (1991). Non-limiting examples of in vitro assays for assessing ADCC activity of a molecule of interest are described in U.S. Pat. Nos. 5,500,362 (see, e.g., Hellstrom, I. et al., Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assays can be used (see, e.g., ACTI for flow cytometry). TM Nonradioactive cytotoxicity assay (Cell Technology, Inc. Mountain View, CA); and CytoTox Non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of the target molecule can be assessed in vivo, for example, in an animal model such as that disclosed in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). C1q binding assays can also be performed to confirm that the antibody cannot bind to C1q and therefore lacks CDC activity. See, for example, C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay can be performed (see, e.g., Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996); Cragg, MS et al., Blood 101: 1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103: 2738-2743 (2004)). FcRn binding and in vivo clearance / half-life assays can also be performed using methods known in the art (see, e.g., Petkova, SB et al., Int'l. Immunol. 18(12): 1759-1769 (2006); WO 2013 / 120929 Al).

[0585] Antibodies with reduced effector function include those with substitutions of one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Pat. No. 6,737,056). Such Fc mutants include those with substitutions at two or more of amino acids 265, 269, 270, 297, and 327, including the so-called "DANA" Fc mutant, in which residues 265 and 297 are substituted with alanine (U.S. Pat. No. 7,332,581).

[0586] Certain antibody variants with improved or decreased binding to FcRs are described. (See, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312; and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001).)

[0587] In certain aspects, the antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC, eg, substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbering of residues).

[0588] In some aspects, the antibody variant comprises an Fc region with one or more amino acid substitutions that reduce FcγR binding, e.g., substitutions at positions 234 and 235 of the Fc region (EU numbering of residues). In one aspect, the substitutions are L234A and L235A (LALA). In some aspects, the antibody variant further comprises a 5-mer amino acid substitution derived from human IgG. 1 D265A and / or P329G in the Fc region of the Fc region. In one aspect, in a 1 In the Fc region, the substitutions are L234A, L235A and P329G (LALA-PG). (See, e.g., WO 2012 / 130831). In another aspect, in a human IgG derived 1 In the Fc region, the substitutions are L234A, L235A and D265A (LALA-DA).

[0589] In some aspects, antibody variants include Fc regions with one or more amino acid substitutions (e.g., substitutions at positions) that improve FcγR binding (and thus improve effector function). In some aspects, antibody variants include Fc regions with at least one amino acid substitution of G236A, I332E, S298A, E333A, K334A, S239D, A330L, F243L, R292P, Y300L, V305I, P396L, L235V, L234Y, L235Q, G236W, S239M, H268D, D270E, K326D, A330M, K334E (see, e.g., Liu et al., Antibodies (Basel) (2020); 9 (4): 64).

[0590] In some aspects, changes are made in the Fc region that result in altered (ie, improved or reduced) CIq binding and / or complement dependent cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164:4178-4184 (2000).

[0591] Antibodies with extended half-life and improved binding to the neonatal Fc receptor (FcRn) are described in US2005 / 0014934 (Hinton et al.), which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)). Those antibodies comprise an Fc region having one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include those having a substitution at one or more of the following Fc region residues: 238, 252, 254, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434, e.g., a substitution to Fc region residue 434 (see, e.g., U.S. Pat. No. 7,371,826; Dall'Acqua, WF et al. J. Biol. Chem. 281 (2006) 23514-23524).

[0592] Fc region residues critical for mouse Fc-mouse FcRn interactions have been identified by site-directed mutagenesis (see, e.g., Dall'Acqua, WF et al. J. Immunol 169 (2002) 5171-5180). The interaction involves residues 1253, H310, H433, N434, and H435 (EU numbering of residues) (Medesan, C. et al., Eur. J. Immunol. 26 (1996) 2533; Firan, M. et al., Int. Immunol. 13 (2001) 993; Kim, JK et al., Eur. J. Immunol. 24 (1994) 542). Residues 1253, H310, and H435 were found to be critical for the interaction of human Fc with murine FcRn (Kim, JK et al., Eur. J. Immunol. 29 (1999) 2819). Studies on the human Fc-human FcRn complex have shown that residues I253, S254, H435 and Y436 are essential for the interaction (Firan, M. et al., Int. Immunol. 13 (2001) 993; Shields, RL et al., J. Biol. Chem. 276 (2001) 6591-6604). Various mutants of residues 248 to 259, 301 to 317, 376 to 382 and 424 to 437 have been reported and examined in Yeung, YA et al. (J. Immunol. 182 (2009) 7667-7671).

[0593] In some aspects, the antibody variant comprises an Fc region with one or more amino acid substitutions that reduce FcRn binding, for example, substitutions at positions 253, and / or 310 and / or 435 of the Fc region (EU numbering of residues). In some aspects, the antibody variant comprises an Fc region with amino acid substitutions at positions 253, 310 and 435. On the one hand, in an Fc region derived from a human IgG1 Fc region, the substitutions are I253A, H310A and H435A. See, e.g., Grevys, A. et al., J. Immunol. 194 (2015) 5497-5508.

[0594] In some aspects, the antibody variant comprises an Fc region with one or more amino acid substitutions that reduce FcRn binding, for example, substitutions at positions 310, and / or 433 and / or 436 of the Fc region (EU numbering of residues). In some aspects, the antibody variant comprises an Fc region with amino acid substitutions at positions 310, 433 and 436. On the one hand, in an Fc region derived from a human IgG1 Fc region, the substitutions are H310A, H433A and Y436A. (See, e.g., WO 2014 / 177460 A1).

[0595] In certain aspects, the antibody variants comprise an Fc region with one or more amino acid substitutions that increase FcRn binding, e.g., substitutions at positions 252, and / or 254 and / or 256 of the Fc region (EU numbering of residues). In certain aspects, the antibody variants comprise an Fc region with amino acid substitutions at positions 252, 254, and 256. In one aspect, in a region derived from human IgG 1 In the Fc region of the Fc region, the substitutions are M252Y, S254T and T256E. For other examples of Fc region variants, see also: Duncan and Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94 / 29351.

[0596] The C-terminus of the heavy chain of the antibody as reported herein can be a complete C-terminus ending with amino acid residue PGK. The C-terminus of the heavy chain can be a shortened C-terminus, in which one or two C-terminal amino acid residues have been removed. In one aspect, the C-terminus of the heavy chain is a shortened C-terminus ending with PG. In one aspect of all aspects reported herein, the antibody comprising a heavy chain including a C-terminal CH3 domain as specified herein, comprises a C-terminal glycine-lysine dipeptide (G446 and K447, EU index numbering of amino acid position). In one aspect of all aspects reported herein, the antibody comprising a heavy chain including a C-terminal CH3 domain as specified herein, comprises a C-terminal glycine residue (G446, EU index numbering of amino acid position). In one aspect of all aspects reported herein, the antibody comprising a heavy chain including a C-terminal CH3 domain as specified herein, comprises a C-terminal proline residue (P445, EU index numbering of amino acid position).

[0597] d) Cysteine ​​engineered antibody variants

[0598] In certain aspects, it may be desirable to generate antibodies with engineered cysteine ​​residues, such as THIOMAB. TM Antibodies, wherein one or more residues of the antibody are replaced by cysteine ​​residues. In a specific embodiment, the substituted residue is present in an accessible site of the antibody. As further described herein, by replacing those residues with cysteine, reactive thiol groups are thus positioned at accessible sites of the antibody, and can be used to conjugate the antibody to other parts (such as drug moieties or linker-drug moieties), to produce immunoconjugates. Cysteine ​​engineered antibodies can be produced as described in, for example, U.S. Patent Nos. 7,521,541, 8,30,930, 7,855,275, 9,000,130 or WO 2016040856.

[0599] e) Antibody derivatives

[0600] In some aspects, the antibodies provided herein may be further modified to include additional non-proteinaceous parts known in the art and readily available. Suitable parts for antibody derivatization include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol / propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers) and dextran or poly-(N-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Due to its stability in water, polyethylene glycol propionaldehyde may have advantages in manufacturing. Polymer may have any molecular weight, and may have or may not have side chains. The number of polymers connected to the antibody may vary, and if more than one polymer is connected, they may be identical or different molecules. Generally, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular property or function of the antibody to be improved, whether the antibody derivative will be used in therapy under defined conditions, etc.

[0601] C. Recombinant Methods and Compositions

[0602] Antibodies as disclosed herein can be produced using recombinant methods and compositions (eg, as described in US 4,816,567). For these methods, one or more isolated nucleic acids encoding the antibodies are provided.

[0603] In the case of a natural antibody or a natural antibody fragment, two nucleic acids are required, one for a light chain or a fragment thereof and one for a heavy chain or a fragment thereof. Such nucleic acid encoding comprises an amino acid sequence of the VL of the antibody and / or comprises an amino acid sequence of the VH of the antibody (e.g., a light chain and / or a heavy chain of the antibody). These nucleic acids can be on the same expression vector or on different expression vectors.

[0604] In the case of certain bispecific antibodies with heterodimeric heavy chains, four nucleic acids are required, one for the first light chain, one for the first heavy chain comprising a first heteromonomeric Fc region polypeptide, one for the second light chain, and one for the second heavy chain comprising a second heteromonomeric Fc region polypeptide. The four nucleic acids may be composed of one or more nucleic acid molecules or expression vectors. Such nucleic acids encode the amino acid sequence of the first VL constituting the antibody and / or the amino acid sequence of the first VH constituting the antibody comprising the first heteromonomeric Fc region and / or the amino acid sequence of the second VL constituting the antibody and / or the amino acid sequence of the second VH constituting the antibody comprising the second heteromonomeric Fc region (e.g., the first light chain and / or the second light chain and / or the first heavy chain and / or the second heavy chain of the antibody). These nucleic acids may be on the same expression vector or on different expression vectors, and typically these nucleic acids are located on two or three expression vectors, i.e., one vector may contain more than one of these nucleic acids. Examples of these bispecific antibodies are (See, e.g., Schaefer, W. et al., PNAS, 108 (2011) 11187-1191). For example, one of the heteromonomer heavy chains comprises a so-called "knob mutation" (T366W, and optionally one of S354C or Y349C), and the other of the heteromonomer heavy chains comprises a so-called "hole mutation" (T366S, L368A and Y407V, and optionally Y349C or S354C) (see, e.g., Carter, P. et al., Immunotechnol. 2 (1996) 73), numbered according to the EU index.

[0605] In one aspect, an isolated nucleic acid encoding an antibody for use in the methods as reported herein is provided.

[0606] In one aspect, a method for preparing an anti-CCR8 antibody is provided, wherein the method comprises culturing a host cell comprising a nucleic acid encoding the antibody as provided above under conditions suitable for expressing the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0607] For recombinant production of anti-CCR8 antibodies, nucleic acids encoding the antibodies (e.g., as described above) are isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acids can be easily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that specifically bind to genes encoding the heavy and light chains of the antibodies), or produced by recombinant methods or obtained by chemical synthesis.

[0608] Suitable host cells for cloning or expressing vectors encoding antibodies include prokaryotic or eukaryotic cells as described herein. For example, antibodies can be produced in bacteria, particularly when glycosylation and Fc effector functions are not required. Regarding the expression of antibody fragments and polypeptides in bacteria, see, for example, US 5,648,237, US 5,789,199 and US 5,840,523. (See also Charlton, KA, in: Methods in Molecular Biology, Vol. 248, Lo, BKC ed., Humana Press, Totowa, NJ (2003), pp. 245-254, describing the expression of antibody fragments in Escherichia coli.) Antibodies can be separated from bacterial cell paste in a soluble fraction after expression, and can be further purified.

[0609] In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are also suitable cloning or expression hosts for vectors encoding antibodies, including fungi and yeast strains whose glycosylation pathways have been "humanized", resulting in the production of antibodies with partially or fully human glycosylation patterns. See Gerngross, TU, Nat. Biotech. 22 (2004) 1409-1414; and Li, H. et al., Nat. Biotech. 24 (2006) 210-215.

[0610] Suitable host cells for expressing glycosylated antibodies also come from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant cells and insect cells. Many baculovirus strains have been identified that can be used with insect cells, particularly for transfecting Spodoptera frugiperda cells.

[0611] Plant cell cultures can also be used as hosts. See, for example, US 5,959,177, US 6,040,498, US 6,420,548, US 7,125,978, and US 6,417,429 (describing PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[0612] Vertebrate cells can also be used as hosts. For example, mammalian cell lines adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line (COS-7) transformed by SV40; human embryonic kidney cell lines (such as 293 or 293T cells described in, for example, Graham, FL et al., J. Gen Virol. 36 (1977) 59-74); small hamster kidney cells (BHK); mouse Sertoli cells (such as TM4 cells described in, for example, Mather, JP, Biol. Reprod. 23 (1980) 243-252); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK); Buffalo rat liver cells (BRL3A); human lung cells (W138); human liver cells (HepG2); mouse mammary tumor (MMT 060562); TRI cells (as described, for example, in Mather, JP et al., Annals N.Y. Acad. Sci. 383 (1982) 44-68); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hams...

Claims

1. A method of treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof, the method comprising administering to the subject a monoclonal antibody that binds to CC motif chemokine receptor 8 (CCR8).

2. The method according to claim 1, wherein: (i) the subject has progressed after at least one available standard therapy; and / or (ii) The subject is a subject for whom all available standard therapies have been proven to be ineffective or for whom the subject is intolerant or has contraindications.

3. The method of claim 1 or 2, wherein the locally advanced, recurrent or metastatic solid tumor is incurable.

4. The method according to any one of claims 1 to 3, wherein the subject is 18 years of age or older.

5. The method according to any one of claims 1 to 4, wherein the locally advanced, recurrent or metastatic solid tumor malignancy is non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), melanoma, triple-negative breast cancer (TNBC), urothelial carcinoma (UC), esophageal cancer, gastric cancer, cervical cancer, renal cell carcinoma (RCC) or hepatocellular carcinoma (HCC). The method of claim 5 , wherein the RCC is clear cell RCC.

7. The method of claim 5, wherein the HNSCC is HNSCC of the oral cavity, oropharynx, hypopharynx, or larynx.

8. The method of claim 5, wherein the locally advanced, recurrent or metastatic solid tumor malignancy is NSCLC.

9. The method of claim 8, wherein the subject's tumor comprises a targetable somatic alteration and the subject has experienced disease progression during or after treatment with a targeted agent or is intolerant to the treatment.

10. The method of claim 9, wherein the targetable somatic alterations include somatic alterations involving epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), ROS proto-oncogene 1 (ROS1), proto-oncogene B-Raf (BRAF) V600E, neurotrophic tyrosine receptor kinase (NTRK), MET proto-oncogene (MET), RET proto-oncogene (RET), or Kirsten rat sarcoma virus (KRAS).

11. The method of claim 5, wherein the melanoma is cutaneous melanoma.

12. The method of claim 11, wherein the subject's tumor comprises a BRAFV600 mutation and the subject has experienced disease progression during or after treatment with one or more serine / threonine protein kinase B-Raf (BRAF) inhibitors and / or one or more mitogen-activated protein kinase (MEK) inhibitors or is intolerant to the treatment.

13. The method of claim 5, wherein the locally advanced, recurrent or metastatic solid tumor malignancy is UC.

14. The method of claim 13, wherein the subject has: (i) histologically confirmed incurable advanced transitional cell carcinoma of the urothelial tract (including the renal pelvis, ureters, bladder and urethra); and / or (ii) Mixed histology, wherein the subject's tumor has a dominant transitional cell pattern.

15. The method of claim 5, wherein the locally advanced, recurrent or metastatic solid tumor malignancy is TNBC.

16. The method of claim 15, wherein TNBC is defined by the American Society of Clinical Oncology-College of American Pathologists guidelines: (i) <1% of tumor cell nuclei are immunoreactive for estrogen receptors and <1% of tumor cell nuclei are immunoreactive for progesterone receptors; and / or (ii) HER2-negative based on immunohistochemistry (IHC) and / or in situ hybridization.

17. The method of any one of claims 1 to 16, wherein the subject is checkpoint inhibitor (CPI) naive.

18. The method of claim 17, wherein the subject's locally advanced, recurrent or metastatic solid tumor malignancy is NSCLC or UC.

19. The method of claim 18, wherein the subject's locally advanced, recurrent or metastatic solid tumor malignancy is UC, wherein the subject is eligible for treatment with cisplatin, and the subject has experienced disease progression during or after treatment with cisplatin or is intolerant to the treatment.

20. The method of any one of claims 17 to 19, wherein the subject has not received prior treatment with a CPI, or wherein the subject has received adjuvant therapy with a CPI that was discontinued at least six months prior to the first administration of the monoclonal antibody that binds to CCR8 to the subject.

21. The method of any one of claims 1 to 16, wherein the subject has experienced a CPI.

22. The method of claim 21, wherein the subject's locally advanced, recurrent or metastatic solid tumor malignancy is NSCLC, HNSCC, melanoma, UC, TNBC, esophageal cancer, gastric cancer, cervical cancer, clear cell RCC, or HCC.

23. The method of claim 21 or 22, wherein the subject has received clinical benefit from treatment comprising a PD-1 axis binding antagonist prior to disease progression.

24. The method of claim 23, wherein the PD-1 axis binding antagonist is an anti-PD-L1 antibody or an anti-PD-1 antibody.

25. The method of claim 23 or 24, wherein the subject has a treatment duration of greater than or equal to 6 months with a therapy comprising a PD-1 axis binding antagonist and / or has a partial response or a complete response as the best objective response.

26. A method according to any one of claims 21 to 25, wherein: (i) the subject has not been treated with a CPI, an immunomodulatory monoclonal antibody, or an immunomodulatory monoclonal antibody-derived therapy within 6 weeks prior to the first administration of the monoclonal antibody that binds to CCR8 to the subject; or (ii) the subject has been previously treated with a PD-1 axis binding antagonist, and the last administration of the PD-1 axis binding antagonist to the subject was at least 3 weeks before the first administration of the monoclonal antibody that binds to CCR8 to the subject.

27. The method of any one of claims 17 to 26, wherein the CPI is a PD-1 axis binding antagonist or a CTLA4 antagonist.

28. The method of claim 27, wherein the PD-1 axis binding antagonist is an anti-PD-L1 antibody or an anti-PD-1 antibody.

29. The method of any one of claims 1 to 28, wherein the monoclonal antibody that binds to CCR8 is administered to the subject in a dosing regimen comprising one or more dosing cycles.

30. The method of claim 29, wherein the one or more dosing cycles comprises a 21 day dosing cycle.

31. The method of claim 30, wherein the monoclonal antibody that binds to CCR8 is administered to the subject on day 1 of each 21-day dosing cycle.

32. The method of any one of claims 29 to 31, wherein the monoclonal antibody that binds to CCR8 is administered to the subject until disease progression or unacceptable toxicity occurs.

33. The method of any one of claims 1 to 32, wherein the monoclonal antibody that binds to CCR8 is administered to the subject at a dose of 2 mg.

34. The method of any one of claims 1 to 33, wherein the monoclonal antibody that binds to CCR8 is administered intravenously to the subject.

35. The method of claim 34, wherein the monoclonal antibody that binds to CCR8 is administered intravenously to the subject by infusion.

36. The method of any one of claims 1 to 35, wherein the monoclonal antibody that binds to CCR8 is administered to the subject as a monotherapy.

37. The method of any one of claims 1 to 35, wherein the monoclonal antibody that binds to CCR8 is administered to the subject in combination with one or more additional therapeutic agents.

38. The method of claim 37, wherein the one or more additional therapeutic agents comprises atezolizumab.

39. The method of claim 38, wherein the atezolizumab is administered to the subject in a dosing regimen comprising one or more dosing cycles.

40. The method of claim 39, wherein the one or more dosing cycles comprises a 21 day dosing cycle.

41. The method of claim 40, wherein the atezolizumab is administered to the subject on day 1 of each 21-day dosing cycle.

42. The method of any one of claims 38 to 41, wherein the atezolizumab is administered to the subject at a dose of 1200 mg.

43. The method of any one of claims 38 to 42, wherein the atezolizumab is administered intravenously to the subject.

44. The method of claim 43, wherein the atezolizumab is administered intravenously to the subject by infusion.

45. The method of any one of claims 1 to 44, wherein a tumor sample from the subject has been determined to have a detectable expression level of PD-L1.

46. ​​The method of claim 45, wherein the tumor sample from the subject has greater than or equal to 1% tumor cells (TC), immune cells (IC), comprehensive positive score (CPS), or tumor proportion score (TPS).

47. The method of any one of claims 38 to 46, wherein the subject has received at least two cycles of the monoclonal antibody that binds to CCR8 prior to administering atezolizumab to the subject.

48. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 29 or SEQ ID NO: 30, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 31, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 32; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 26, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 27, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

28.

49. The method of claim 48, wherein the monoclonal antibody that binds to CCR8 binds to CCR8 independently of sulfation of CCR8.

50. The method of claim 48 or 49, wherein the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:

106.

51. The method of any one of claims 48 to 50, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35 to 47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 to 52; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

52. The method of any one of claims 48 to 51, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35 to 47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52.

53. The method of any one of claims 48 to 52, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 47; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 48; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

54. The method of any one of claims 48 to 53, wherein the monoclonal antibody that binds to CCR8 comprises: a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:47; and a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:

48.

55. The method of any one of claims 48 to 54, wherein the VL comprises a V4M mutation, a P43A mutation, a F46L mutation, a C90Q mutation, or a combination thereof (according to Kabat numbering).

56. The method of any one of claims 48 to 55, wherein the VH comprises a G49S mutation, a K71R mutation, a S73N mutation, or a combination thereof (according to Kabat numbering).

57. The method of any one of claims 48 to 56, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 55 and a light chain amino acid sequence of SEQ ID NO:

56.

58. The method of any one of claims 48 to 56, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 60 and a light chain amino acid sequence of SEQ ID NO:

56.

59. The method of any one of claims 48 to 56, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 111 and a light chain amino acid sequence of SEQ ID NO:

56.

60. The method of any one of claims 48 to 56, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 113 and a light chain amino acid sequence of SEQ ID NO:

56.

61. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 35 to 47 and a VL sequence selected from the group consisting of SEQ ID NOs: 48 to 52.

62. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 47 and a VL sequence of SEQ ID NO:

48.

63. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:7; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:1, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:2, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

3.

64. The method of claim 63, wherein the monoclonal antibody that binds to CCR8 binds to CCR8 independently of sulfation of CCR8.

65. The method of claim 63 or 64, wherein the monoclonal antibody that binds to CCR8 binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:

106.

66. The method of any one of claims 63 to 65, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 to 25; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

67. The method of any one of claims 63 to 66, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence selected from the group consisting of SEQ ID NOs: 10 to 21 and a VL sequence selected from the group consisting of SEQ ID NOs: 22 to 25.

68. The method of any one of claims 63 to 67, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 21; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 24; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

69. The method of any one of claims 63 to 68, wherein the monoclonal antibody that binds to CCR8 comprises: a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:21; and a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:

24.

70. The method of any one of claims 63 to 69, wherein the VL comprises a Y2I mutation (according to Kabat numbering).

71. The method of any one of claims 63 to 70, wherein the VH comprises an S73N mutation, a V78L mutation, a T76N mutation, a F91Y mutation, and a P105Q mutation, or a combination thereof (according to Kabat numbering).

72. The method of any one of claims 63 to 71, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 57 and a light chain amino acid sequence of SEQ ID NO:

58.

73. The method of any one of claims 63 to 71, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 61 and a light chain amino acid sequence of SEQ ID NO:

58.

74. The method of any one of claims 63 to 71, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 112 and a light chain amino acid sequence of SEQ ID NO:

58.

75. The method of any one of claims 63 to 71, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 114 and a light chain amino acid sequence of SEQ ID NO:

58.

76. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 21 and a VL sequence of SEQ ID NO:

24.

77. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 83, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 84, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 85; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 73, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 74, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

75.

78. The method of claim 77, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 95; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 94; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

79. The method of claim 77 or 78, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 95 and a VL sequence of SEQ ID NO:

94.

80. The method of any one of claims 77 to 79, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 101 and a light chain amino acid sequence of SEQ ID NO:

100.

81. The method of any one of claims 77 to 79, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 115 and a light chain amino acid sequence of SEQ ID NO:

100.

82. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 86 or SEQ ID NO: 87, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 89; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 76, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 77, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

78.

83. The method of claim 82, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 97; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 96; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

84. The method of claim 82 or 83, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 97 and a VL sequence of SEQ ID NO:

96.

85. The method of any one of claims 82 to 84, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 103 and a light chain amino acid sequence of SEQ ID NO:

102.

86. The method of any one of claims 82 to 84, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 116 and a light chain amino acid sequence of SEQ ID NO:

102.

87. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 90 or SEQ ID NO: 91, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 92, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 93; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 79, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 80, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

81.

88. The method of claim 87, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 99; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 98; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

89. The method of claim 87 or 88, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 99 and a VL sequence of SEQ ID NO:

98.

90. The method of any one of claims 87 to 89, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 105 and a light chain amino acid sequence of SEQ ID NO:

104.

91. The method of any one of claims 87 to 90, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 117 and a light chain amino acid sequence of SEQ ID NO:

104.

92. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 binds to CCR8 independently of sulfation of CCR8.

93. The method of claim 92, wherein the antibody binds to an epitope comprising one or more of amino acid residues 2 to 6 of SEQ ID NO:

106.

94. The method of claim 92, wherein the antibody binds to an epitope comprising one or more of amino acid residues 91 to 104 and 172 to 193 of SEQ ID NO:

106.

95. The method of any one of claims 1 to 47, wherein the monoclonal antibody that binds to CCR8 comprises: a heavy chain variable domain (VH) comprising (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 65 or SEQ ID NO: 66, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and a light chain variable domain (VL) comprising (d) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 62, (e) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 63, and (f) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:

64.

96. The method of claim 95, wherein the monoclonal antibody that binds to CCR8 comprises a sequence selected from the group consisting of: (a) a VH sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO:70; (b) a VL sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of SEQ ID NO: 69; and (c) a VH sequence as defined in (a) and a VL sequence as defined in (b).

97. The method of claim 95 or 96, wherein the monoclonal antibody that binds to CCR8 comprises a VH sequence of SEQ ID NO: 70 and a VL sequence of SEQ ID NO:

69.

98. The method of any one of claims 95 to 97, wherein the monoclonal antibody that binds to CCR8 comprises a heavy chain amino acid sequence of SEQ ID NO: 72 and a light chain amino acid sequence of SEQ ID NO:

71.

99. The method of any one of claims 1 to 98, wherein the monoclonal antibody that binds to CCR8 is a human antibody.

100. The method of any one of claims 1 to 98, wherein the monoclonal antibody that binds to CCR8 is a humanized antibody.

101. The method of any one of claims 1 to 100, wherein the monoclonal antibody that binds to CCR8 is a chimeric antibody.

102. The method of any one of claims 1 to 101, wherein the monoclonal antibody that binds to CCR8 is an antibody fragment that binds to CCR8.

103. The method of any one of claims 1 to 101, wherein the monoclonal antibody that binds to CCR8 is a full-length antibody.

104. The method of claim 103, wherein the monoclonal antibody that binds to CCR8 is a full-length IgG1 antibody.

105. The method of any one of claims 1 to 104, wherein the monoclonal antibody that binds to CCR8 comprises an IgG1 constant domain comprising the amino acid sequence of SEQ ID NO:53 or SEQ ID NO:

59.

106. The method of any one of claims 1 to 105, wherein the monoclonal antibody that binds to CCR8 comprises a kappa constant domain comprising the amino acid sequence of SEQ ID NO:

54.

107. The method of any one of claims 1 to 106, wherein the monoclonal antibody that binds to CCR8 is present at about 1×10 -12 M is about 1×10 -11 The binding affinity (K d ) binds to CCR8.

108. The method of any one of claims 1 to 107, wherein the CCR8 is human CCR8.

109. The method of any one of claims 1 to 108, wherein the monoclonal antibody that binds to CCR8 is defucosylated.

110. The method of claim 109, wherein the proportion of defucosylation is between about 80% and about 95%.

111. The method of any one of claims 1 to 110, wherein regulatory T cells present in the tumor microenvironment of the locally advanced, recurrent or metastatic solid tumor malignancy are depleted.

112. The method of any one of claims 1 to 111, wherein regulatory T cells outside the tumor microenvironment of the locally advanced, recurrent or metastatic solid tumor malignancy are depleted.

113. The method of any one of claims 1 to 112, wherein the subject is a human.

114. A monoclonal antibody that binds to CCR8 for use in treating a locally advanced, recurrent or metastatic solid tumor malignancy in a subject in need thereof.

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