Anti-cancer therapy using a combination of Anti-CCR8

Abstract

The present disclosure relates to methods of treating cancer in a subject by administering to the subject an effective amount of an anti-CCR8 antibody, a chemotherapeutic agent (e.g., cis-platinum, gemcitabine, docetaxel), and a PD1 inhibitor or a PD-L1 inhibitor. In some embodiments, the chemotherapeutic agent is administered at a lower dose than in a standard care chemotherapeutic regimen that does not comprise an anti-CCR8 antibody.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 382,386, filed on November 4, 2022, under 35 U.S.C. § 119(e), which is hereby incorporated by reference in its entirety for all purposes. Technical Field

[0003] The present disclosure relates to methods of treating cancer in a subject by co - administering to the subject an effective amount of an anti - CCR8 antibody, a chemotherapeutic agent (e.g., cisplatin, gemcitabine, docetaxel), and a PD1 inhibitor or a PD - L1 inhibitor. In some embodiments, the chemotherapeutic agent is co - administered at a lower dose than in a standard - of - care chemotherapy regimen that does not include an anti - CCR8 antibody.

[0004] Sequence Listing

[0005] This application contains a sequence listing, which is submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML copy was created on October 12, 2023, named 1456 - US - NP_SL.xml, and is 126,949 bytes in size. Background Art

[0006] The lack of effector T - cell activity in solid tumors can be attributed to inhibitory mechanisms exploited by regulatory T cells (Tregs) in the tumor microenvironment. A prominent problem in the art is how to selectively deplete intratumoral Tregs to avoid severe autoimmunity triggered by systemic depletion.

[0007] Chemokine (C - C motif) receptor 8 (CCR8) belongs to the G - protein - coupled receptor (GPCR) family. CCR8 is highly expressed on the surface of tumor - infiltrating Tregs, but not on peripheral Tregs and effector T cells. CCR8 - targeting antibodies that result in rapid depletion of intratumoral Tregs in mouse models and human ex vivo systems are currently used clinically as monotherapies and in combination with checkpoint inhibitors.

[0008] There remains a need for additional combination therapies to improve the efficacy of anti - CCR8 - antibody - based anti - cancer therapies. Summary of the Invention

[0009] In one aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising co-administering to the subject an effective amount of: (i) an anti-CCR8 antibody; (ii) a chemotherapeutic agent; and (iii) a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody); wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody.

[0010] In some embodiments, the chemotherapeutic agent is co-administered at a lower dose than in a standard of care chemotherapeutic regimen that does not include the anti-CCR8 antibody.

[0011] In another aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising co-administering to the subject an effective amount of: (i) an anti-CCR8 antibody; (ii) a chemotherapeutic agent; and (iii) optionally a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody); wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity; wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody; and wherein the chemotherapeutic agent is co-administered at a lower dose than in a standard of care chemotherapeutic regimen that does not include the anti-CCR8 antibody.

[0012] In some embodiments, the chemotherapeutic agent is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is a plurality of chemotherapeutic agents. In some embodiments, the chemotherapeutic agent is selected from the group consisting of: platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin.

[0013] In some embodiments, the chemotherapeutic agent comprises a platinum complex. In some embodiments, the platinum complex is selected from the group consisting of: carboplatin, cisplatin, and oxaliplatin. In some embodiments, the chemotherapeutic agent comprises gemcitabine. In some embodiments, the chemotherapeutic agent comprises a taxane. In some embodiments, the taxane is docetaxel. In some embodiments, the dose of the chemotherapeutic agent is 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 50% or less, 40% or less, 30% or less, or 20% or less of the dose of the chemotherapeutic agent administered in a standard of care regimen that does not include the anti-CCR8 antibody.

[0014] In some embodiments, the cancer comprises a solid tumor.

[0015] In some embodiments, the cancer comprises tumor-infiltrating Treg cells that express CCR8. In some embodiments, CCR8 is expressed on the surface of Treg cells at less than 10,000 copies / cell (as determined by fluorescence-activated cell sorting (FACS) and / or flow cytometry).

[0016] In some embodiments, the cancer is selected from the group consisting of: breast cancer, colorectal cancer, head and neck cancer, lung cancer, ovarian cancer, gastric cancer, gastric adenocarcinoma, and thymoma. In some embodiments, the cancer is selected from the group consisting of: endometrial adenocarcinoma, colorectal cancer, ovarian cancer, vaginal squamous cell carcinoma, endometrial adenocarcinoma, colorectal cancer, melanoma (e.g., cutaneous melanoma), pancreatic cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), uterine leiomyosarcoma, cholangiocarcinoma, adenoid cystic carcinoma, cervical cancer, renal cell carcinoma (RCC), anal cancer, esophagogastric junction (EGJ) adenocarcinoma, and gastric adenocarcinoma. In some embodiments, the cancer is selected from the group consisting of: head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer (NSCLC), gastric adenocarcinoma, EGJ adenocarcinoma, and colorectal cancer (CRC) (e.g., microsatellite stable (MSS) mCRC). In some embodiments, the cancer is selected from the group consisting of: breast cancer, pancreatic cancer, and lung cancer. In some embodiments, the breast cancer is selected from triple-negative breast cancer (TNBC), HR + / HER 2- breast cancer or HR + / HER2 低 breast cancer. In some embodiments, the pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). In some embodiments, the cancer is lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC). In some embodiments, the lung cancer is NSCLC. In some embodiments, the cancer is metastatic.

[0017] In some embodiments, the cancer is ovarian cancer and the co-administered chemotherapeutic agent is selected from the group consisting of: 5-fluorouracil, albumin-bound paclitaxel, hexamethylmelamine, anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, exemestane, gemcitabine, ifosfamide, irinotecan, letrozole, leuprolide acetate, liposomal doxorubicin, megestrol acetate, melphalan, olaparib, oxaliplatin, paclitaxel, pazopanib, pemetrexed, tamoxifen, topotecan, vinorelbine, and any combination thereof.

[0018] In some embodiments, the cancer is HNSCC and the co-administered chemotherapeutic agent is selected from the group consisting of afatinib, bleomycin, capecitabine, carboplatin, cetuximab, cisplatin, docetaxel, fluorouracil, gemcitabine, hydroxyurea, methotrexate, nivolumab, paclitaxel, vinorelbine, and any combination thereof.

[0019] In some embodiments, the cancer is gastric adenocarcinoma and the co-administered chemotherapeutic agent is selected from the group consisting of capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, mitomycin, oxaliplatin, paclitaxel, and any combination thereof.

[0020] In some embodiments, the cancer is esophageal-gastric junction (EGJ) adenocarcinoma and the co-administered chemotherapeutic agent is selected from the group consisting of capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, oxaliplatin, paclitaxel, and any combination thereof.

[0021] In some embodiments, the cancer is colorectal cancer and the co-administered chemotherapeutic agent is selected from the group consisting of capecitabine, cetuximab, fluorouracil, irinotecan, leucovorin, oxaliplatin, panitumumab, ziv-aflibercept, and any combination thereof.

[0022] In some embodiments, the cancer is breast cancer and the co-administered chemotherapeutic agent is selected from the group consisting of albumin-bound paclitaxel, anastrozole, atezolizumab, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, epirubicin, everolimus, exemestane, fluorouracil, fulvestrant, gemcitabine, ixabepilone, lapatinib, letrozole, methotrexate, mitoxantrone, paclitaxel, pegylated liposomal doxorubicin, pertuzumab, tamoxifen, toremifene, trastuzumab, vinorelbine, and any combination thereof.

[0023] In some embodiments, the breast cancer is TNBC and the co-administered chemotherapeutic agent is selected from the group consisting of cyclophosphamide, docetaxel, doxorubicin, epirubicin, fluorouracil, paclitaxel, and any combination thereof.

[0024] In some embodiments, the cancer is lung cancer and the co-administered chemotherapeutic agent is selected from the group consisting of afatinib, albumin-bound paclitaxel, alectinib, cabozantinib, carboplatin, cisplatin, crizotinib, dabrafenib, docetaxel, erlotinib, etoposide, gemcitabine, paclitaxel, pemetrexed, vandetanib, vemurafenib, vinblastine, vinorelbine, and any combination thereof.

[0025] In some embodiments, the lung cancer is SCLC and the co-administered chemotherapeutic agent is selected from the group consisting of: 5-fluorouracil, albumin-bound paclitaxel, altretamine, anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, exemestane, gemcitabine, ifosfamide, irinotecan, letrozole, leuprolide acetate, liposomal doxorubicin, megestrol acetate, melphalan, olaparib, oxaliplatin, paclitaxel, pazopanib, pemetrexed, tamoxifen, topotecan, vinorelbine, and any combination thereof.

[0026] In some embodiments, the cancer is a blood cancer that expresses CCR8. In some embodiments, the blood cancer is selected from the group consisting of: T-cell adult acute lymphoblastic leukemia, T-cell pediatric acute lymphoblastic leukemia, lymphoblastic lymphoma, acute lymphoblastic leukemia, cutaneous T-cell lymphoma (CTCL), T-cell acute lymphoblastic leukemia, adult T-cell leukemia / lymphoma, T-cell lymphoblastic leukemia / lymphoma, and anaplastic large cell lymphoma. In some embodiments, the blood cancer is CTCL.

[0027] In some embodiments, the subject is a human. In some embodiments, the subject has not received treatment. In some embodiments, the subject has received one or more courses of anti-cancer treatment, and optionally the cancer has progressed during one or more courses of anti-cancer treatment. In some embodiments, the anti-cancer treatment is selected from the group consisting of: surgery, radiotherapy, hormone therapy, targeted anti-cancer agents, chemotherapeutic agents, immunotherapy, and antibody-drug conjugates (ADCs). In some embodiments, the chemotherapeutic agent is selected from the group consisting of: platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin. In some embodiments, the platinum complex is selected from the group consisting of: carboplatin, cisplatin, and oxaliplatin. In some embodiments, the taxane is docetaxel. In some embodiments, the immunotherapy comprises an anti-PD-1 antibody or an anti-PD-L1 antibody. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is selected from the group consisting of: pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, tislelizumab, rivulizumab, btilimab, toripalimab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and sepacilimab. In some embodiments, the immunotherapy further comprises an anti-TIGIT antibody. In some embodiments, the anti-TIGIT antibody is selected from the group consisting of: tirzepatide, vibostolimab, donanemab, AB308, AK127, BMS-986207, or eptinezumab.

[0028] In some embodiments, the anti-CCR8 antibody comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 13, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 14, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 16, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 17; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 25, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 26, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 27, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 29; (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 37, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 39, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 40, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 41; (d) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 48, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 49, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 50, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 51, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 52, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 53; or (e) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 60, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 61, 72 or 78, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 62, 73 or 79, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 63, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 64, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 65;or (f) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 84 or 100, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 85, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 86, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 87, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 88, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 89. In some embodiments, the anti-CCR8 antibody comprises: (a) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 68 or 74, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 69 or 75; or (b) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 92 or 96, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 93 or 97. In some embodiments, the anti-CCR8 antibody comprises: (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 68 or 74, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 69 or 75; or (b) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 92 or 96, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 93 or 97. In some embodiments, the anti-CCR8 antibody is a monoclonal antibody. In some embodiments, the anti-CCR8 antibody is a humanized antibody. In some embodiments, the anti-CCR8 antibody is a full-length antibody. In some embodiments, the anti-CCR8 antibody is an IgG1 or IgG3 antibody. In some embodiments, the anti-CCR8 antibody comprises: (a) a heavy chain (HC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 70 or 76, and a light chain (LC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 71 or 77;or (b) a heavy chain (HC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 94 or 98, and a light chain (LC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 95 or 99. In some embodiments, the anti-CCR8 antibody comprises: (a) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 70 or 76, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 71 or 77;or (b) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 94 or 98, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 95 or 99. In some embodiments, the anti-CCR8 antibody is a fucosylation-free antibody. In some embodiments, the anti-CCR8 antibody comprises a heavy chain constant region mutation at one or more positions selected from L234, L235, G236, S239, F243, H268, D270, R292, S298, Y300, V305, K326, A330, I332, E333, K334 and P396. In some embodiments, the anti-CCR8 antibody comprises a heavy chain constant region mutation selected from S239D, S239M, F243L, H268D, D270E, R292P, S298A, Y300L, V305I, K326D, A330L, A330M, I332E, E333A, K334A, K334E and P396L. In some embodiments, the anti-CCR8 antibody comprises a heavy chain constant region mutation selected from F243L / R292P / Y300L / V305I / P396L, S239D / I332E, S239D / I332E / A330L, S298A / E333A / K334A, L234Y / L235Q / G236W / S239M / H268D / D270E / S298A and D270E / K326D / A330M / K334E. In some embodiments, the anti-CCR8 antibody inhibits the binding of CCL1 to CCR8. In some embodiments, the anti-CCR8 antibody is selected from the group consisting of: BMS-986340 (Bristol Myers Squibb), LM-108 (LaNova Medicines), S-531011 (Shionogi), FPA157 (Five Prime, Amgen), IPG-7236 (Immunophage Biomedical), ICP-B05 (InnoCare Pharma Tech), SRF-114 (Surface Oncology), HBM1022 (HarbourBioMed), HFB1011 (HiFiBio), BAY-3375968 (Bayer), IO-1 (Oncurious), ZL-1218 (Zai Lab), GB2101 (Genor) and PSB-114 (Sound Biologics).;

[0029] In some embodiments, the co-administered PD-1 inhibitor or PD-L1 inhibitor is an anti-PD1 antibody or an anti-PD-L1 antibody. In some embodiments, the co-administered anti-PD-1 antibody or anti-PD-L1 antibody is selected from the group consisting of: pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, tislelizumab, rivulizumab, btilimumab, toripalimab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and sepantronium. In some embodiments, the co-administered PD-1 inhibitor or PD-L1 inhibitor is a small molecule. In some embodiments, the small molecule PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of: CA-170, GS-4224, GS-4416, INCB99280, INCB99318, and lazertinib.

[0030] In some embodiments, the methods provided herein further comprise co-administering to a subject one or more additional therapeutic agents.

[0031] In another aspect, the present disclosure provides an anti-CCR8 antibody for use in combination with a chemotherapeutic agent and an anti-PD-1 antibody or an anti-PD-L1 antibody in a method of treating cancer, wherein the method comprises co-administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the anti-PD1 antibody or the anti-PD-L1 antibody, wherein the anti-CCR8 antibody has antibody-dependent cell-mediated cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody.

[0032] In another aspect, the present disclosure provides an anti-CCR8 antibody for use in combination with a chemotherapeutic agent and optionally a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) in a method of treating cancer, wherein the method comprises co-administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the PD-1 inhibitor or the PD-L1 inhibitor (e.g., the anti-PD1 antibody or the anti-PD-L1 antibody), wherein the anti-CCR8 antibody has antibody-dependent cell-mediated cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody, and wherein the chemotherapeutic agent is administered at a lower dose than in a standard-of-care chemotherapy regimen that does not include an anti-CCR8 antibody agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1Shows the tumor growth curves in a syngeneic mouse breast cancer model (4T1). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, low-dose chemotherapy (cisplatin), or a combination thereof.

[0034] Figure 2 Shows the tumor growth curves in a syngeneic mouse pancreatic cancer model (Panc02). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, low-dose chemotherapy (gemcitabine), or a combination thereof.

[0035] Figure 3 Shows the tumor growth curves in a syngeneic mouse melanoma model (B16F10). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, low-dose chemotherapy (gemcitabine), or a combination thereof.

[0036] Figure 4A and Figure 4B Shows a bar graph illustrating tumor-infiltrating lymphocytes from a syngeneic mouse melanoma model (B16F10). Mice were each administered a single dose of a control antibody, gemcitabine (“low-dose SOC chemotherapeutic agent”), and / or an anti-CCR8 antibody and the frequency of regulatory T cells (Treg; Figure 3 A) or effector T cells ( Figure 3 B) was quantified. The results of a t-test are shown, where **** is p < 0.0001, *** is p < 0.001, ** is p < 0.01 and * is p < 0.1.

[0037] Figure 5 Shows the tumor growth curves in a syngeneic mouse melanoma model (B16F10). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, an anti-PD-1 antibody, or a combination thereof.

[0038] Figure 6 Shows the tumor growth curves in a syngeneic mouse lung adenocarcinoma model (LLC). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, low-dose chemotherapy (docetaxel), or a combination thereof.

[0039] Figure 7 Shows the tumor growth curves in a syngeneic mouse lung adenocarcinoma model (LLC). Mice were each administered a single dose of a control antibody, an anti-CCR8 antibody, low-dose chemotherapy (docetaxel), an anti-PD-1 antibody, an anti-CCR8 antibody / chemotherapy combination, an anti-PD-1 antibody / chemotherapy combination, or an anti-CCR8 antibody / anti-PD-1 antibody / chemotherapy combination.

[0040] Figure 8 Shows the illustration of Figure 7Bar graph of tumor volume of the LLC model treatment cohort shown in [Figure] at day 15. Detailed implementation manners

[0041] Definition

[0042] Unless otherwise defined, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by one of ordinary skill in the art. Additionally, unless the context otherwise requires or clearly indicates, singular terms shall include the plural and plural terms shall include the singular. In case of any conflict in definitions between various sources or references, the definitions provided herein shall prevail.

[0043] It should be understood that the embodiments of the invention described herein include "consisting of" and / or "consisting essentially of" embodiments. Unless otherwise indicated, as used herein, the singular forms "a", "an", and "the" include plural referents. The use of the term "or" herein does not imply that the alternatives are mutually exclusive.

[0044] Unless explicitly stated or understood by those skilled in the art, in this application, the use of "or" means "and / or". In the context of multiple dependent claims, the use of "or" refers to more than one preceding independent or dependent claim.

[0045] As understood by those skilled in the art, the reference herein to a "about" value or parameter includes (and describes) embodiments directed to that value or parameter itself. For example, the description of "about X" includes the description of "X".

[0046] As used herein, "CCR8" and "C-C chemokine receptor type 8" and "chemokine receptor 8" refer to any native CCR8 produced by the expression and processing of CCR8 in a cell. Unless otherwise indicated, the term includes CCR8 from any vertebrate source, which vertebrates include mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats). The term also includes naturally occurring variants of CCR8, such as splice variants or allelic variants. The amino acid sequence of an exemplary human CCR8 protein is shown as SEQ ID NO: 101 (UniProt identifier P51685). The amino acid sequence of an exemplary mouse CCR8 protein is shown as SEQ ID NO: 102 (UniProt identifier P56484). The amino acid sequence of an exemplary cynomolgus monkey CCR8 protein is shown as SEQ ID NO: 103 (UniProt identifier G7NYJ2).

[0047] As used herein, "CCL1" and "C-C motif chemokine 1" refer to any native CCR1 produced by the expression and processing of CCR1 in a cell. Unless otherwise indicated, the term includes CCR1 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats). The term also includes naturally occurring variants of CCR1, such as splice variants or allelic variants. The amino acid sequence of an exemplary human CCR1 protein is shown as SEQ ID NO: 2 (UniProt identifier P22362.1). The exemplary mature CCR1 protein comprises amino acids 24-96 of SEQ ID NO: 2.

[0048] As used herein, a "7-B16 antibody" is understood to be any antibody that binds to CCR8 and comprises: (i) a heavy chain containing SEQ ID NO: 82 and a light chain containing SEQ ID NO: 83, (ii) a heavy chain variable region containing SEQ ID NO: 80 and a light chain variable region containing SEQ ID NO: 81, or (iii) HCDR1, HCDR2, and HCDR3 containing SEQ ID NO: 84, 85, and 86, respectively, and LCDR1, LCDR2, and LCDR3 containing SEQ ID NO: 87, 88, and 89, respectively; and any chimeric, human, or humanized version of any of the foregoing (i), (ii), or (iii). In some embodiments, a "7-B16 antibody" may be used to specifically refer to an antibody comprising a heavy chain of SEQ ID NO: 82 and a light chain of SEQ ID NO: 83.

[0049] As used herein, the term "anti-PD-1 antibody" or "anti-PD-L1 antibody" refers to: a) an antibody that binds to programmed cell death protein 1 (PD-1, CD279; NCBI Gene ID: 5133) or programmed death ligand 1 (PD-L1, CD274; NCBI Gene ID: 29126); and b) an antibody that inhibits PD-1 / PD-L1 interaction and the PD-1 / PD-L1 pathway. The PD-1 / PD-L1 pathway and its role in cancer immunotherapy are described, for example, in Salmaninejad et al., J. Cell Physiol (2019) 234 (10): 16824-16837. Anti-PD-1 antibodies or anti-PD-L1 antibodies that can be used in the methods provided herein include, for example, pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, tislelizumab, rivulizumab, btilimab, toripalimab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and sepaclimab. In some embodiments, the anti-PD-1 antibody is sepaclimab.

[0050] The term "specifically binds" to an antigen or epitope is a term well known in the art, and methods for determining such specific binding are also well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular cell or substance more frequently, more rapidly, for a longer duration, and / or with greater affinity than it does with an alternative cell or substance. An antibody "specifically binds" or "preferentially binds" to a target if it binds to the target with greater affinity, avidity, more readily, and / or for a longer duration than it does to other substances. For example, an antibody that specifically or preferentially binds to a CCR8 epitope is an antibody that binds to that epitope with greater affinity, avidity, more readily, and / or for a longer duration than it binds to other CCR8 epitopes or non-CCR8 epitopes. It should also be understood from reading this definition that, for example, an antibody (or portion or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Generally, but not necessarily, reference to binding implies preferential binding. "Specificity" refers to the ability of a binding protein to selectively bind an antigen.

[0051] As used herein, "substantially pure" refers to a substance that is at least 50% pure (i.e., free of contaminants), more preferably at least 90% pure, more preferably at least 95% pure, still more preferably at least 98% pure, and most preferably at least 99% pure.

[0052] As used herein, the term "epitope" refers to a site on a target molecule (e.g., an antigen such as a protein, nucleic acid, carbohydrate, or lipid) to which an antigen-binding molecule (e.g., an antibody, an antibody fragment, or a scaffold protein containing an antibody-binding region) binds. Epitopes typically include the chemically reactive surface groups of a molecule such as amino acids, polypeptides, or sugar side chains, and have specific three-dimensional structural features as well as specific charge features. Epitopes can be formed by both contiguous residues and / or juxtaposed non-contiguous residues (e.g., amino acids, nucleotides, sugars, lipid moieties) of the target molecule. Epitopes formed by contiguous residues (e.g., amino acids, nucleotides, sugars, lipid moieties) typically remain upon exposure to denaturing solvents, whereas epitopes formed by tertiary folding typically are lost upon treatment with denaturing solvents. Epitopes can include, but are not limited to, at least 3, at least 5, or 8-10 residues (e.g., amino acids or nucleotides). In some examples, the length of an epitope is less than 20 residues (e.g., amino acids or nucleotides), less than 15 residues, or less than 12 residues. If two antibodies exhibit competitive binding to an antigen, they can bind to the same epitope within the antigen. In some embodiments, an epitope can be identified by a certain minimum distance from the CDR residues on an antigen-binding molecule. In some embodiments, an epitope can be identified by the above distance and is further limited to those residues involved in a bond (e.g., a hydrogen bond) between an antibody residue and an antigen residue. Epitopes can also be identified by various scans, e.g., alanine or arginine scans can indicate one or more residues with which an antigen-binding molecule can interact. Unless explicitly stated, a set of residues identified as an epitope does not exclude other residues from being part of the epitope for a particular antibody. Instead, the presence of such a set designates a minimum series (or group of species) of the epitope. Thus, in some embodiments, a set of residues identified as an epitope designates the minimum epitope associated with an antigen, rather than a unique list of the residues of the epitope on the antigen.

[0053] The term "antibody" as used herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific (such as bispecific T cell engagers) and trispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0054] The term antibody includes but is not limited to fragments capable of binding an antigen, such as Fv, single-chain Fv (scFv), Fab, Fab’, bis-scFv, sdAb (single domain antibody), and (Fab’) 2 (including chemically linked F(ab’)2 ). Papain digestion of an antibody produces two identical antigen-binding fragments (referred to as "Fab" fragments, each with a single antigen-binding site), and a residual "Fc" fragment (the name reflects the ability to crystallize readily). Pepsin treatment yields F(ab’) 2 fragments that have two antigen-binding sites and are still able to cross-link antigens. The term antibody also includes, but is not limited to, chimeric antibodies, humanized antibodies, and antibodies from various species such as mouse, human, cynomolgus monkey, etc. In addition, for all antibody constructs provided herein, variants having sequences from other organisms are also contemplated. Thus, if the human version of an antibody is disclosed, one of ordinary skill in the art will understand how to convert the human sequence-based antibody into sequences of mouse, rat, cat, dog, horse, etc. Antibody fragments also include any orientation of single-chain scFv, tandem bis-scFv, diabodies, tandem tris-sdcFv, minibodies, etc. Antibody fragments also include nanobodies (sdAb, an antibody having a single monomeric domain such as a pair of heavy-chain variable domains without a light chain). In some embodiments, an antibody fragment may be referred to as being of a particular species (e.g., human scFv or mouse scFv). This refers to the sequence of at least part of the non-CDR region and not the source of the construct.

[0055] The term "monoclonal antibody" refers to an antibody of a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. Thus, a monoclonal antibody sample can bind to the same epitope on an antigen. The modifier "monoclonal" indicates the character of the antibody obtained from a substantially homogeneous population of antibodies and should not be construed as requiring that the antibody be made by any particular method. For example, monoclonal antibodies can be prepared by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or can be prepared by recombinant DNA methods such as those described in U.S. Patent No. 4,816,567. Monoclonal antibodies can also be isolated from phage libraries generated using, for example, the techniques described in McCafferty et al., 1990, Nature 348:552-554.

[0056] The term "CDR" refers to the complementarity determining regions as defined by one or more methods of identification by those skilled in the art. In some embodiments, the CDRs may be defined according to any Chothia numbering scheme, Kabat numbering scheme, a combination of Kabat and Chothia, AbM definition, Contact definition, and / or a combination of Kabat, Chothia, AbM, and / or Contact definitions. Exemplary CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) occur at amino acid residues 24-34 of L1, 50-56 of L2, 89-97 of L3, 31-35B of H1, 50-65 of H2, and 95-102 of H3. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)). The AbM definition may include, for example, CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) at amino acid residues 24-34 of L1, 50-56 of L2, 89-97 of L3, H26-H35B of H1, 50-58 of H2, and 95-102 of H3. The Contact definition may include, for example, CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) at amino acid residues 30-36 of L1, 46-55 of L2, 89-96 of L3, 30-35 of H1, 47-58 of H2, and 93-101 of H3. The Chothia definition may include, for example, CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) at amino acid residues 24-34 of L1, 50-56 of L2, 89-97 of L3, 26-32...34 of H1, 52-56 of H2, and 95-102 of H3. CDRs as shown in any one or more of the figures may also be provided. In addition to V HIn addition to CDR1 in, CDRs generally contain amino acid residues that form hypervariable loops. Each CDR in an antibody can be designated by its appropriate numbering and chain type, including but not limited to being designated as: a) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3; b) CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3; c) LCDR-1, LCDR-2, LCDR-3, HCDR-1, HCDR-2, and HCDR-3; or d) LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3; and so on. The term "CDR" is also used herein to encompass HVR or "hypervariable region", including hypervariable loops. Exemplary hypervariable loops occur at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3). (Chothia and Lesk, J. Mol. Biol. 196:901-917(1987).)

[0057] As used herein, the term "heavy chain variable region" refers to a region that contains at least three heavy chain CDRs. In some embodiments, the heavy chain variable region includes three CDRs and at least FR2 and FR3. In some embodiments, the heavy chain variable region contains at least heavy chain HCDR1, framework (FR) 2, HCDR2, FR3, and HCDR3. In some embodiments, the heavy chain variable region further contains at least a portion of FR1 and / or at least a portion of FR4.

[0058] As used herein, the term "heavy chain constant region" refers to a region that contains at least three heavy chain constant regions C H 1, C H 2, and C H 3. Of course, unless otherwise specified, non-functional alterations, deletions, and alterations within the domain are encompassed within the term "heavy chain constant region". Non-limiting exemplary heavy chain constant regions include γ, δ, and α. Non-limiting exemplary heavy chain constant regions also include ε and µ. Each heavy chain constant region corresponds to an antibody isotype. For example, an antibody containing a γ constant region is an IgG antibody, an antibody containing a δ constant region is an IgD antibody, and an antibody containing an α constant region is an IgA antibody. In addition, an antibody containing a µ constant region is an IgM antibody, and an antibody containing an ε constant region is an IgE antibody. Certain isotypes can also be further divided into subclasses. For example, IgG antibodies include but are not limited to IgG1 (containing γ 1 constant region), IgG2 (containing γ 2 constant region), IgG3 (containing γ 3 constant region), and IgG4 (containing γ 4Constant region) antibodies; IgA antibodies include, but are not limited to, IgA1 (comprising α 1 constant region) and IgA2 (comprising α 2 constant region) antibodies; and IgM antibodies include, but are not limited to, IgM1 and IgM2.

[0059] As used herein, the term "heavy chain" refers to a polypeptide comprising at least one heavy chain variable region, with or without a leader sequence. In some embodiments, the heavy chain comprises at least a portion of the heavy chain constant region. As used herein, the term "full-length heavy chain" refers to a polypeptide comprising a heavy chain variable region and a heavy chain constant region, with or without a leader sequence.

[0060] As used herein, the term "light chain variable region" refers to a region comprising at least three light chain CDRs. In some embodiments, the light chain variable region comprises three CDRs and at least FR2 and FR3. In some embodiments, the light chain variable region comprises at least light chain LCDR1, framework (FR) 2, LCDR2, FR3, and LCDR3. For example, the light chain variable region may comprise light chain CDR1, framework (FR) 2, CDR2, FR3, and CDR3. In some embodiments, the light chain variable region further comprises at least a portion of FR1 and / or at least a portion of FR4.

[0061] As used herein, the term "light chain constant region" refers to a region comprising the light chain constant domain C L of. Non-limiting exemplary light chain constant regions include λ and κ. Of course, unless otherwise specified, non-functional alterations, deletions, and alterations within the domain are encompassed within the scope of the term "light chain constant region".

[0062] As used herein, the term "light chain" refers to a polypeptide comprising at least one light chain variable region, with or without a leader sequence. In some embodiments, the light chain comprises at least a portion of the light chain constant region. As used herein, the term "full-length light chain" refers to a polypeptide comprising a light chain variable region and a light chain constant region, with or without a leader sequence.

[0063] For purposes herein, "receptor human framework" is a framework comprising the amino acid sequence of a light chain variable domain (V L ) framework or a heavy chain variable domain (V H ) framework of a derived human immunoglobulin framework or a human consensus framework as defined below. A receptor human framework derived from a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence thereof, or the receptor human framework may contain amino acid sequence variations. In some embodiments, the number of amino acid changes is 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer. In some embodiments, V LThe acceptor human framework is identical in sequence to the V L human immunoglobulin framework sequence or the human consensus framework sequence.

[0064] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (such as an antibody) and its binding partner (such as an antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (K D ). It can be measured by conventional methods known in the art (such as ELISA K D , KinExA, biolayer interferometry (BLI) and / or surface plasmon resonance devices (such as BIAcore ® devices), including those described herein).

[0065] As used herein, the term "K D " refers to the equilibrium dissociation constant of the antibody-antigen interaction.

[0066] In some embodiments, the "K ® ", "K ® ", "Kd" or "Kd value" of an antibody is measured at 25 °C using a BIACORE D -2000 or BIACORE d -3000 (BIAcore, Inc., Piscataway, N.J.) with an immobilized antigen CM5 chip of approximately 10 response units (RU). Briefly, the carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) is activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. The antigen is diluted to 5 μg / ml (about 0.2 μM) with 10 mM sodium acetate (pH 4.8), and then injected at a flow rate of 5 μL / min to achieve coupling of approximately 10 response units (RU) of protein. After injecting the antigen, 1 M ethanolamine is injected to block unreacted groups. For kinetic measurements, serial dilutions of the polypeptide (such as a full-length antibody) are injected in PBS with 0.05% TWEEN-20 ™ surfactant (PBST) at a flow rate of approximately 25 μL / min at 25 °C. The association rate (k ® ) and dissociation rate (k on ) are calculated by simultaneously fitting the association and dissociation sensorgrams using a simple one-to-one Langmuir binding model (BIACORE off evaluation software version 3.2). The equilibrium dissociation constant (K d ) is calculated as k off / kon Ratio. See, e.g., Chen et al., J. Mol. Biol. 293:865 - 881 (1999). If, as determined by the surface plasmon resonance described above, the association rate exceeds 10 6 M -1 s -1 , then the association rate can be determined by using fluorescence quenching techniques that measure the increase or decrease in fluorescence emission intensity of 20 nM antigen - specific antibody in PBS (pH 7.2) at 25°C in the presence of increasing concentrations of antigen (excitation = 295 nm; emission = 340 nm, band - pass 16 nm), as measured in a spectrofluorometer, such as a stopped - flow equipped spectrophotometer (Aviv Instruments) or an 8000 series SLM - AMINCO ™ spectrophotometer (ThermoSpectronic).

[0067] The term "biological activity" refers to any one or more biological properties of a molecule (whether naturally occurring as found in vivo, or provided or enabled by recombinant means). Biological properties include, but are not limited to, binding cytokines, inducing cell proliferation, inhibiting cell growth, inducing other cytokines, inducing apoptosis, and enzymatic activity. In some embodiments, the biological activity of CCR8 includes anti - apoptotic activity, cell chemotaxis, immunosuppressive function, and the ability to polarize cells towards various cell differentiation pathways.

[0068] As used herein, a "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 at least a portion of the remaining heavy and / or light chain is derived from a different source or species. In some embodiments, a chimeric antibody refers to an antibody that contains at least one variable region from a first species (such as mouse, rat, cynomolgus monkey, etc.) and at least one constant region from a second species (such as human, cynomolgus monkey, etc.). In some embodiments, a chimeric antibody contains at least one murine variable region and at least one human constant region. In some embodiments, a chimeric antibody contains at least one cynomolgus monkey variable region and at least one human constant region. In some embodiments, all variable regions of the chimeric antibody are from the first species and all constant regions of the chimeric antibody are from the second species. As described above, chimeric constructs can also be functional fragments.

[0069] As used herein, a "humanized antibody" refers to an antibody in which at least one amino acid in the framework region of a non - human variable region has been replaced with the corresponding amino acid from a human variable region. In some embodiments, a humanized antibody contains at least one human constant region or a fragment thereof. In some embodiments, a humanized antibody is an antibody fragment, such as Fab, scFv, (Fab') 2etc. The term humanized also refers to chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab') 2 or other antigen-binding sequences of an antibody) in the form of a non-human (e.g., murine) antibody containing a minimal sequence of a non-human immunoglobulin. A humanized antibody includes a human immunoglobulin (recipient antibody) in which the residues of the complementarity-determining regions (CDRs) from the recipient are replaced with residues of CDRs having the desired specificity, affinity, and capacity from a non-human species (such as a mouse, rat, or rabbit) (donor antibody). In some cases, the framework region (FR) residues of the human immunoglobulin are replaced with corresponding non-human residues. In addition, a humanized antibody can contain residues that are not present in the recipient antibody or in the input CDR or framework sequences, but the inclusion of such residues is for further improving and optimizing antibody performance. Generally, a humanized antibody can contain substantially all of at least one, usually two variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of a non-human immunoglobulin, and all or substantially all of the framework (FR) regions are framework regions of a human immunoglobulin consensus sequence. In some embodiments, a humanized antibody can also contain at least a portion of an immunoglobulin constant region or domain (Fc) (usually a constant region or domain of a human immunoglobulin). Other forms of humanized antibodies have one or more CDRs (CDR L1, CDR L2, CDR L3, CDR H1, CDR H2, and / or CDR H3) that are altered relative to the original antibody, which are also referred to as one or more CDRs "derived" from one or more CDRs of the original antibody. It will be understood that a humanized sequence can be identified by its primary sequence and does not necessarily represent the process by which the antibody is produced.

[0070] As used herein, a "CDR-grafted antibody" refers to a humanized antibody in which one or more complementarity-determining regions (CDRs) of a first (non-human) species have been grafted onto the framework region (FR) of a second (human) species.

[0071] As used herein, "human antibody" encompasses antibodies produced in humans, antibodies produced in non-human animals containing human immunoglobulin genes such as XenoMouse ®Antibodies generated in mice and antibodies selected using in vitro methods such as phage display (Vaughan et al., 1996, Nature Biotechnology, 14:309-314; Sheets et al., 1998, Proc. Natl. Acad. Sci. (USA) 95:6157-6162; Hoogenboom and Winter, 1991, J. Mol. Biol., 227:381; Marks et al., 1991, J. Mol. Biol., 222:581), wherein the antibody library is based on human immunoglobulin sequences. The term "human antibody" refers to a genus of sequences that are human sequences. Thus, the term does not refer to the method of generating the antibody, but rather to the genus of related sequences.

[0072] A "functional Fc region" has the "effector functions" of a native sequence Fc region. Exemplary "effector functions" include Fc receptor binding; C1q binding; CDC; ADCC; phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require the combination of the Fc region with a binding domain (e.g., an antibody variable domain) and can be evaluated using a variety of assays.

[0073] A "native sequence Fc region" comprises an amino acid sequence that is the same as the amino acid sequence of a naturally occurring Fc region. Native sequence human Fc regions include native sequence human IgG1 Fc region (non-A and A allotypes); native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region and their naturally occurring variants.

[0074] A "variant Fc region" comprises an amino acid sequence that is different from the amino acid sequence of a native sequence Fc region due to at least one amino acid modification. In some embodiments, a "variant Fc region" comprises an amino acid sequence that is different from the amino acid sequence of a native sequence Fc region due to at least one amino acid modification but retains at least one effector function of the native sequence Fc region. In some embodiments, the variant Fc region has at least one amino acid substitution compared to the native sequence Fc region or the Fc region of the parental polypeptide, e.g., about 1 to about 10 amino acid substitutions, preferably about 1 to about 5 amino acid substitutions in the native sequence Fc region or the Fc region of the parental polypeptide. In some embodiments, a variant Fc region herein will have at least about 80% sequence identity with the native sequence Fc region and / or the Fc region of the parental polypeptide, at least about 90% sequence identity therewith, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity therewith.

[0075] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. In some embodiments, FcγR is a native human FcR. In some embodiments, the FcR is an FcR (γ receptor) that binds IgG antibodies and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of those receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain. (See, e.g., Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcR is reviewed, for example, in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med., 126:330-41 (1995). The term "FcR" as used herein encompasses other FcRs, including those yet to be identified.

[0076] The term "Fc receptor" or "FcR" also includes the neonatal receptor FcRn, 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)) and the regulation of immunoglobulin homeostasis. Methods for measuring binding to FcRn are known (see, e.g., Ghetie and Ward., Immunol. Today 18(12):592-598 (1997); Ghetie et al., Nature Biotechnology, 15(7):637-640 (1997); Hinton et al., J. Biol. Chem. 279(8):6213-6216 (2004); WO 2004 / 92219 (Hinton et al.).

[0077] "Effector function" refers to the biological activities attributable to the Fc region of an antibody that vary with antibody isotype. Examples of antibody effector functions include: Clq 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 receptors); and B cell activation.

[0078] "Human effector cells" are white blood cells that express one or more FcRs and perform effector functions. In some embodiments, the cells express at least FcγRIII and perform ADCC effector functions. Examples of human white blood cells that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils. Effector cells can be isolated from natural sources, such as from blood.

[0079] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages), enabling these cytotoxic effector cells to specifically bind to target cells carrying the antigen and subsequently kill the target cells with cytotoxins. Primary cell NK cells that mediate ADCC express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. The expression of FcRs on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay can be performed, such as those described in U.S. Patent No. 5,500,362 or 5,821,337 or U.S. Patent No. 6,737,056 (Presta). Effector cells that can be used in such assays include PBMCs and NK cells. Alternatively or in addition, the ADCC activity of a molecule of interest can be evaluated in vivo, for example, in an animal model (such as the animal model disclosed in Clynes et al., Proc. Natl. Acad. Sci. (USA) 95:652-656 (1998)). Additional polypeptide variants having altered Fc region amino acid sequences (polypeptides having variant Fc regions) and increased or decreased ADCC activity are described, for example, in U.S. Patent No. 7,923,538 and U.S. Patent No. 7,994,290.

[0080] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to an antibody (of the appropriate subclass) bound to its cognate antigen. To assess complement activation, a CDC assay can be performed, for example, as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Polypeptide variants having an altered amino acid sequence in the Fc region (polypeptides having a variant Fc region) and increased or decreased C1q binding ability are described, for example, in U.S. Patent No. 6,194,551 B1, U.S. Patent No. 7,923,538, U.S. Patent No. 7,994,290, and WO 1999 / 51642. See also, for example, Idusogie et al., J. Immunol. 164: 4178-4184 (2000).

[0081] A polypeptide variant having an "altered" FcR binding affinity or ADCC activity is a polypeptide variant having enhanced or diminished FcR binding activity and / or ADCC activity as compared to a parental polypeptide or as compared to a polypeptide comprising a native sequence Fc region. A polypeptide variant that "exhibits" "increased binding" to an FcR binds at least one FcR with a higher affinity than the parental polypeptide. A polypeptide variant that "exhibits" "decreased binding" to an FcR binds at least one FcR with a lower affinity than the parental polypeptide. Such variants that exhibit decreased binding to an FcR as compared to the native sequence IgG Fc region may have little or no appreciable binding to the FcR, e.g., 0-20% binding to the FcR as compared to the native sequence IgG Fc region.

[0082] A polypeptide variant that "more effectively mediates antibody-dependent cell-mediated cytotoxicity (ADCC) in the presence of human effector cells" as compared to a parental antibody is a polypeptide variant that more effectively mediates ADCC in vitro or in vivo when the amounts of the polypeptide variant and the parental antibody used in the assay are substantially the same. Typically, such variants will be identified using an in vitro ADCC assay as disclosed herein, but other assays or methods for determining ADCC activity, e.g., in animal models, etc., are also contemplated.

[0083] As used herein, the terms "substantially similar" or "substantially identical" denote a high enough degree of similarity between two or more values such that one of ordinary skill in the art would consider the differences between the two or more values to have little or no biological and / or statistical significance within the context of the biological characteristic being measured by the values. In some embodiments, two or more substantially similar values differ by no more than about 5%, 10%, 15%, 20%, 25%, or 50%.

[0084] As used herein, the phrase "substantially different" indicates a high enough degree of difference between two numerical values such that one of ordinary skill in the art would consider the difference between the two values to be statistically significant within the context of the biological characteristic being measured by the values. In some embodiments, two substantially different numerical values differ by more than about 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0085] As used herein, the phrase "significantly reduced" indicates a high enough degree of reduction between a numerical value and a reference numerical value such that one of ordinary skill in the art would consider the difference between the two values to be statistically significant within the context of the biological characteristic being measured by the values. In some embodiments, a significantly reduced numerical value is reduced by more than about 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100% compared to the reference value.

[0086] The term "leader sequence" refers to an amino acid residue sequence located at the N-terminus of a polypeptide that facilitates the secretion of the polypeptide from mammalian cells. The leader sequence can be cleaved when the polypeptide is exported from mammalian cells, thereby forming a mature protein. Leader sequences can be natural or synthetic, and they can be heterologous or homologous to the protein to which they are attached.

[0087] A "native sequence" polypeptide comprises a polypeptide having the same amino acid sequence as a naturally occurring polypeptide. Thus, a native sequence polypeptide can have the amino acid sequence of a naturally occurring polypeptide from any mammal. Such native sequence polypeptides can be isolated from nature or can be produced by recombinant or synthetic methods. The term "native sequence" polypeptide specifically encompasses naturally occurring truncated forms or secreted forms of a polypeptide (e.g., extracellular domain sequences), naturally occurring variant forms of a polypeptide (e.g., alternative splicing forms), and naturally occurring allelic variants of a polypeptide.

[0088] A polypeptide "variant" means a bioactive polypeptide that has at least about 80% amino acid sequence identity to a native sequence polypeptide after aligning the sequences and, if necessary, introducing gaps to obtain the maximum percentage of sequence identity and without considering any conservative substitutions as part of the sequence identity. Such variants include, for example, polypeptides in which one or more amino acid residues are added or deleted at the N-terminus or C-terminus of the polypeptide. In some embodiments, the variant will have at least about 80% amino acid sequence identity. In some embodiments, the variant will have at least about 90% amino acid sequence identity. In some embodiments, the variant has at least about 95% amino acid sequence identity to the native sequence polypeptide.

[0089] As used herein, "percent amino acid sequence identity (%)" and "homology" with respect to a peptide, polypeptide or antibody sequence are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a specific peptide or polypeptide sequence after aligning the sequences and introducing gaps if necessary to obtain the maximum percentage of sequence identity and without considering any conservative substitutions as part of the sequence identity. The alignment for determining the percent amino acid sequence identity can be achieved in various ways within the skill in the art, such as using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGN TM (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for measuring the alignment, including any algorithms necessary to achieve the maximum alignment over the full length of the sequences being compared.

[0090] Amino acid substitutions can include, but are not limited to, the replacement of one amino acid in a polypeptide with another. Exemplary conservative substitutions are shown in Table 1. Amino acid substitutions can be introduced into the antibody of interest, and products having the desired activity (e.g., retained / improved antigen binding, reduced immunogenicity or improved ADCC or CDC) can be screened.

[0091] Table 1

[0092]

[0093] Amino acids can be grouped according to common side chain characteristics:

[0094] (1) Hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

[0095] (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

[0096] (3) Acidic: Asp, Glu;

[0097] (4) Basic: His, Lys, Arg;

[0098] (5) Residues affecting strand orientation: Gly, Pro;

[0099] (6) Aromatic: Trp, Tyr, Phe.

[0100] Non-conservative substitutions would require the conversion of a member of one of these classes to another.

[0101] The term "vector" is used to describe a polynucleotide that can be engineered to contain a cloned polynucleotide or a polynucleotide that can be propagated in a host cell. A vector can include one or more of the following elements: an origin of replication, one or more regulatory sequences that regulate the expression of a polypeptide of interest (e.g., a promoter and / or enhancer), and / or one or more selectable marker genes (e.g., an antibiotic resistance gene and a gene that can be used for a colorimetric assay, such as β-galactosidase). The term "expression vector" refers to a vector that is used for expressing a polypeptide of interest in a host cell.

[0102] "Host cell" refers to a cell that can be or has been a recipient of a vector or an isolated polynucleotide. A host cell can be a prokaryotic cell or a eukaryotic cell. Exemplary eukaryotic cells include mammalian cells, such as primate or non-primate animal cells; fungal cells, such as yeast; plant cells; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NSO cells, PER.C6 ® cells (Crucell) and 293 and CHO cells, and their derivatives, such as 293-6E and DG44 cells, respectively. A host cell includes the progeny of a single host cell, and the progeny may not be identical to the original parental cell (in terms of morphology or genomic DNA complementation) due to natural, accidental, or deliberate mutations. A host cell includes a cell transfected in vivo with a polynucleotide provided herein.

[0103] As used herein, the term "isolated" refers to a molecule that has been separated from at least some of the components with which it is normally associated in nature or produced. For example, a polypeptide is referred to as "isolated" when it is separated from at least some of the components of the cell that produces it. When a polypeptide is secreted by a cell after expression, the supernatant containing the polypeptide that is physically separated from the cell that produces it is considered to "isolate" the polypeptide. Similarly, a polynucleotide is referred to as "isolated" when it is not part of the larger polynucleotide (e.g., genomic DNA or mitochondrial DNA in the case of a DNA polynucleotide) with which it is normally associated in nature, or when it is separated from at least some of the components of the cell that produces it (e.g., in the case of an RNA polynucleotide). Thus, a DNA polynucleotide contained in a vector within a host cell can be referred to as "isolated".

[0104] The terms "individual" or "subject" are used interchangeably herein and refer to an animal, such as a mammal. In some embodiments, methods of treating a mammal are provided, which mammals include, but are not limited to, humans, rodents, apes, felines, canines, equines, bovines, pigs, sheep, goats, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets. In some examples, the "individual" or "subject" refers to an individual or subject in need of treatment for a disease or disorder. In some embodiments, the subject being treated can be a patient, designating the fact that the subject has been identified as having a disorder related to the treatment or being at sufficient risk of developing the disorder.

[0105] As used herein, "disease" or "disorder" refers to a condition that requires and / or would benefit from treatment.

[0106] As used herein, "cancer" and "tumor" are interchangeable terms that refer to any abnormal growth or proliferation of cells or tissues in an animal. As used herein, the terms "cancer" and "tumor" encompass solid cancers and blood / lymph cancers, as well as malignant, pre-malignant, and benign growths, such as dysplasia. Examples of cancers include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific non-limiting examples of such cancers include squamous cell carcinoma, small cell lung cancer, pituitary carcinoma, esophageal cancer, astrocytoma, soft tissue sarcoma, non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, peritoneal cancer, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer (kidney cancer / renal cancer), liver cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, brain cancer, endometrial cancer, testicular cancer, cholangiocarcinoma, gallbladder cancer, gastric cancer, melanoma, mesothelioma, and various types of head and neck cancers. In some embodiments, blood / lymph cancers are referred to as "blood cancers". Non-limiting exemplary blood cancers include B-cell and T-cell mixed leukemia, B-cell lymphoma, chronic myeloid leukemia (CML), chronic myelomonocytic leukemia, diffuse large B-cell lymphoma (DLBC), lymphoma, mantle cell lymphoma (MCL), multiple myeloma, myelodysplastic syndrome (MDS), myeloproliferative disorders, peripheral T-cell lymphoma, T-cell leukemia, acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), CLL / SLL, mature T-cell and NK-cell lymphoma, follicular lymphoma, acute lymphocytic leukemia (ALL), T-cell acute lymphocytic leukemia (TALL), T-cell adult acute lymphocytic leukemia, T-cell childhood acute lymphocytic leukemia, lymphoblastic lymphoma, cutaneous T-cell lymphoma (CTCL), adult T-cell leukemia / lymphoma (ATLL), T-cell lymphoblastic leukemia / lymphoma (TLLL), angioimmunoblastic T-cell lymphoma (ATCL), hepatosplenic T-cell lymphoma (HTCL), peripheral T-cell lymphoma not otherwise specified (PTCL NOS), Burkitt lymphoma (BL), chronic myelomonocytic leukemia (CMML), extranodal NK / T-cell lymphoma (NKTCL), primary effusion lymphoma (PEL), acute lymphocytic leukemia / acute myeloid leukemia (ALL, AML), histiocytic lymphoma (HL), marginal zone lymphoma (MZL), B-cell acute lymphocytic leukemia, and anaplastic large cell lymphoma (ALCL).

[0107] As used herein, "treatment" is a method for obtaining a beneficial or desired clinical outcome. "Treatment" as used herein encompasses any administration or application of a therapeutic agent for the treatment of a disease in a mammal, including a human. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, any one or more of the following: alleviation of one or more symptoms, reduction in the severity of a disease, prevention or delay of the spread of a disease (e.g., metastasis, e.g., to the lung or lymph nodes), prevention or delay of the recurrence of a disease, delay or slowing of disease progression, improvement of the disease state, inhibition of a disease or disease progression, inhibition or slowing of a disease or its progression, arrest of its development, and remission (whether partial or complete). "Treatment" also encompasses reducing the pathological consequences of a proliferative disease. The methods provided herein encompass any one or more of these treatment aspects. Consistent with the foregoing, the term treatment does not require 100% removal of all aspects of the disorder.

[0108] "Improvement" means the alleviation or amelioration of one or more symptoms as compared to not administering an anti-CCR8 antibody. "Improvement" also includes shortening or reducing the duration of the symptoms.

[0109] In the context of cancer, the term "treatment" includes any one or all of the following: inhibition of cancer cell growth, inhibition of cancer cell replication, reduction of the overall tumor burden, and improvement of one or more symptoms associated with the disease.

[0110] As used herein, the term "regulatory T cell" (also referred to as "Treg" or "Treg cell" or "suppressor T cell") is a subset of T cells that are immunosuppressive and generally inhibit or downregulate the induction and proliferation of effector T cells. Tregs express CD4, FOXP3, and CD25 (the α chain of the IL-2 receptor). Based on the expression levels of Foxp3 and the cell surface molecules CD25 and CD45RA, human Foxp3+CD4+ T cells are divided into three subfractions. The Foxp3hiCD45RA−CD25hi and Foxp3loCD45RA+CD25lo phenotypes correspond to suppressive Treg cells, whereas the Foxp3loCD45RA−CD25lo fraction labels activated T effector (Teff) cells that have no suppressive activity. In addition, compared to Treg cells in healthy subjects, Treg cells from cancer patients are typically characterized by a unique expression profile of chemokine receptors such as CCR4, CXCR4, and CCR5, which promotes their migration into tumors in response to the corresponding chemokine ligands from the tumor microenvironment. See, e.g., Liu et al., FEBS J. (2016) 283(14):2731-48, and Miyara et al., Immunity (2009) 30, 899–911.

[0111] "Conventional T cells" or "Tconv" are a population of T cells that are typically CD4 positive (i.e., CD4+), but differ from Tregs in that Tconv are typically FoxP3 negative (i.e., FoxP3-).

[0112] The term "biological sample" means an amount of material from a living or formerly living thing. Such materials include, but are not limited to, blood (e.g., whole blood), plasma, serum, urine, amniotic fluid, synovial fluid, endothelial cells, white blood cells, monocytes, other cells, organs, tissues, bone marrow, lymph nodes, and spleen.

[0113] The term "control" refers to a composition that is known to not contain an analyte ("negative control") or contains an analyte ("positive control"). A positive control may contain a known concentration of the analyte. "Control", "positive control", and "calibrator" are used interchangeably herein and refer to a composition that contains a known concentration of an analyte. A "positive control" can be used to establish assay performance characteristics and is a useful indicator of the integrity of a reagent (e.g., an analyte).

[0114] "Predetermined cut-off value" and "predetermined level" generally refer to an assay cut-off value used to evaluate a diagnostic / prognostic / therapeutic efficacy outcome by comparing an assay result to the predetermined cut-off value / level, where the predetermined cut-off value / level has been associated or correlated with various clinical parameters (e.g., disease severity, progression / non-progression / improvement, etc.). While the present disclosure may provide exemplary predetermined levels, it is well known that cut-off values can vary depending on the nature of the immunoassay (e.g., the antibodies used, etc.). Further, applying the disclosure herein to other immunoassays to obtain immunoassay-specific cut-off values for those other immunoassays is within the skill of the ordinary artisan in the art. Although the exact value of the predetermined cut-off value / level may vary between assays, the correlations (if any) as described herein are generally applicable.

[0115] The terms "inhibition" or "inhibit" refer to a decrease or cessation of any phenotypic trait or a decrease or cessation of the incidence, degree, or likelihood of that trait. "Decrease" or "inhibit" means a decrease, reduction, or prevention of activity, function, and / or amount compared to a reference. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 20% or greater. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 50% or greater. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 75%, 85%, 90%, 95% or greater. In some embodiments, the above amounts are inhibited or reduced over a period of time relative to a control dose (such as a placebo) over the same period of time. As used herein, "reference" refers to any sample, standard, or level for comparison purposes. A reference can be obtained from healthy and / or non-diseased samples. In some examples, the reference can be obtained from untreated samples. In some examples, the reference is obtained from non-diseased and untreated samples of an individual subject. In some embodiments, the reference is obtained from one or more healthy individuals who are not the subject or patient.

[0116] As used herein, "delaying the development of a disease" means postponing, hindering, slowing, retarding, stabilizing, suppressing, and / or deferring the development of a disease (such as cancer). The delay can have different time lengths, depending on the history of the disease and / or the individual to be treated. It will be apparent to those skilled in the art that a sufficient or significant delay can actually encompass prevention, since the individual does not develop the disease. For example, the development of advanced cancer, such as metastasis, can be delayed.

[0117] As used herein, "prevention" includes providing prophylaxis against the occurrence or recurrence of a disease in a subject who may be susceptible to the disease but has not been diagnosed with the disease. Unless otherwise specified, the terms "reduce", "inhibit", or "prevent" do not imply or require complete prevention at all times.

[0118] As used herein, "inhibiting" a function or activity means reducing the function or activity when compared to conditions that are otherwise the same except for the condition or parameter of interest or alternatively when compared to another condition. For example, an antibody that inhibits tumor growth reduces the rate of tumor growth compared to the rate of tumor growth in the absence of the antibody.

[0119] A "therapeutically effective amount" of a substance / molecule, agonist or antagonist can vary depending on factors such as the individual's disease state, age, sex and weight, as well as the ability of the substance / molecule, agonist or antagonist to elicit a desired response in the individual. A therapeutically effective amount is also the amount at which the therapeutic beneficial effects outweigh any toxic or detrimental effects of the substance / molecule, agonist or antagonist. A therapeutically effective amount can be delivered in one or more administrations. A therapeutically effective amount refers to the amount that is effective in achieving the desired therapeutic and / or prophylactic outcome at the necessary dosage and for the necessary period of time.

[0120] A prophylactically effective amount refers to the amount that is effective in achieving the desired prophylactic outcome at a certain dosage and for the necessary period of time. Typically, but not necessarily, since prophylactic doses are used in a subject before or at an early stage of a disease, a prophylactically effective amount will be less than a therapeutically effective amount.

[0121] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to formulations that are in a form that permits the biological activity of the active ingredient to be effective and that do not contain additional components that are unacceptably toxic to the subject to whom the formulation will be administered. Such formulations can be sterile.

[0122] A "pharmaceutically acceptable carrier" is a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material, formulation aid or carrier conventional in the art that is used with a therapeutic agent and together constitutes a "pharmaceutical composition" for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosages and concentrations employed and is compatible with the other components of the formulation. A pharmaceutically acceptable carrier is suitable for the formulation used.

[0123] A "sterile" formulation is sterile or substantially free of live microorganisms and their spores.

[0124] "Chimeric antigen receptor T cell therapy" or "CAR-T therapy" refers to a therapeutic agent comprising T cells genetically modified to express a receptor that recognizes an antigen expressed by tumor cells. The antigen can be an antigen specifically expressed by the tumor or an antigen expressed by both cancer cells and healthy tissue. In some embodiments, the CAR-T therapy is adoptive CAR-T therapy, in which patient T cells are removed and modified to express a chimeric antigen receptor and then returned to the patient. See, e.g., Dai et al., 2016, J Natl Cancer Inst, 108(7): djv439, doi: 10.1093 / jnci / djv439; Gill et al., 2015, Blood Rev, pii: S0268-960X(15)00080-6, doi: 10.1016 / j.blre.2015.10.003; Gill et al., 2015, Immunol Rev, 263(1):68-89. doi: 10.1111 / imr.12243.

[0125] "Administering" in "combination" with one or more additional therapeutic agents includes simultaneous (parallel) and sequential or successive administration in any order.

[0126] The term "simultaneously" as used herein refers to the administration of two or more therapeutic agents, where at least a portion of the administrations overlap in time or where the administration of one therapeutic agent falls within a short time period relative to the administration of another therapeutic agent. For example, two or more therapeutic agents are administered at time intervals not exceeding about a specified number of minutes.

[0127] The term "sequentially" as used herein refers to the administration of two or more therapeutic agents, where the administration of one or more agents continues after interruption of the administration of one or more other agents, or where the administration of one or more agents begins before the administration of one or more other agents. For example, the administrations of two or more therapeutic agents are administered at time intervals exceeding about a specified number of minutes.

[0128] As used herein, "co-administering" refers to administering one form of treatment in addition to another form of treatment. Thus, "co-administering" refers to administering one form of treatment before, during, or after the administration of another form of treatment to a subject.

[0129] The term "package insert" is used to refer to the instructions typically included in the commercial packaging of a therapeutic product that contain information about indications, usage, dosage, administration, combination therapy, contraindications and / or warnings regarding the use of such therapeutic product.

[0130] "Article" means any article (e.g., a package or container) or kit that contains at least one reagent, such as a drug for treating a disease or disorder (e.g., cancer) or a probe for specifically detecting a biomarker described herein. In some embodiments, the article or kit is promoted, distributed, or sold as a unit for performing the methods described herein.

[0131] The terms "label" and "detectable label" mean a moiety that is linked to an antibody or its analyte such that the reaction (e.g., binding) between the members of the specific binding pair is detectable. The labeled member of the specific binding pair is referred to as "detectable-labeled". Thus, the term "labeled binding protein" refers to a protein that incorporates a label that provides for the identification of the binding protein. In some embodiments, the label is a detectable marker that can generate a signal detectable by visual or instrumental means, such as an amino acid incorporated with a radioactive label or a polypeptide linked to a biotinyl moiety that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or enzyme activity detectable by optical or colorimetric methods). Examples of labeling of polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g., 3 H, 14 C, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho or 153 Sm); chromogens, fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent markers; biotin groups; a predetermined polypeptide epitope recognized by a second reporter (e.g., leucine zipper pair sequences, binding sites of second antibodies, metal-binding domains, epitope tags); and magnetic agents such as gadolinium chelates. Representative examples of labels commonly used in immunoassays include moieties that produce light (e.g., acridinium compounds) and moieties that produce fluorescence (e.g., fluorescein). In this regard, the moiety itself may not be detectably labeled but may become detectable upon reaction with another moiety.

[0132] The term "conjugate" refers to an antibody that is chemically linked to a second chemical moiety, such as a therapeutic agent or a cytotoxic agent. The term "agent" includes chemical compounds, mixtures of chemical compounds, biological macromolecules, or extracts made from biological materials. In some embodiments, the therapeutic agent or cytotoxic agent includes, but is not limited to, pertussis toxin, paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dactinomycin, mitoxantrone, plicamycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and their analogs or homologs. When used in the context of an immunoassay, the conjugated antibody can be an antibody that serves as a detectable label for a detection antibody.

[0133] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0134] All references cited herein, including patent applications, patent publications, and Genbank accession numbers, are incorporated herein by reference as if each individual reference were specifically and individually indicated to be incorporated by reference in its entirety.

[0135] The techniques and procedures described or referred to herein are generally well understood and commonly used by those skilled in the art using conventional methodologies, such as the widely utilized methodologies described in the following references: Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd Edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. CURRENT PROTOCOLS IN MOLECULAR BIOLOGY (edited by F. M. Ausubel et al. (2003)); METHODS IN ENZYMOLOGY series (Academic Press, Inc.): PCR 2: A PRACTICAL APPROACH (edited by M. J. MacPherson, B. D. Hames and G. R. Taylor (1995)) Harlow and Lane editors (1988) ANTIBODIES, A LABORATORY MANUAL, and ANIMAL CELL CULTURE (edited by R. I. Freshney (1987)); Oligonucleotide Synthesis (edited by M. J. Gait, 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (edited by J. E. Cellis, 1998) Academic Press; Animal Cell Culture (edited by R. I. Freshney, 1987); Introduction to Cell and Tissue Culture (J. P. Mather and P. E. Roberts, 1998) Plenum Press; Cell and Tissue Culture Laboratory Procedures (edited by A. Doyle, J. B. Griffiths and D. G. Newell, 1993 - 8) J. Wiley and Sons; Handbook of Experimental Immunology (edited by D. M. Weir and C. C. Blackwell); Gene Transfer Vectors for Mammalian Cells (J. M. Miller and M. P.Edited by Calos, 1987); PCR: The Polymerase Chain Reaction (Edited by Mullis et al., 1994); Current Protocols in Immunology (Edited by J. E. Coligan et al., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C. A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (Edited by D. Catty., IRL Press, 1988 - 1989); Monoclonal Antibodies: A Practical Approach (Edited by P. Shepherd and C. Dean, Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1999); The Antibodies (Edited by M. Zanetti and J. D. Capra, Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (Edited by V. T. DeVita et al., J. B. Lippincott Company, 1993); and their updated versions.

[0136] Therapeutic method

[0137] The present disclosure is at least partially based on the recognition that intratumoral Tregs expressing CCR8 can be bound by anti-CCR8 antibodies and selectively depleted by natural killer (NK) cells via antibody-dependent cellular cytotoxicity (ADCC). Without wishing to be bound by any theory, it is further believed that simultaneously targeting tumor cells with chemotherapy can trigger immunogenic cell death and result in the uptake of tumor antigens by antigen-presenting cells (APCs). In the absence of the immunosuppressive effects of Tregs, effector T cells (Teffs) can be effectively activated and result in improved tumor killing. Additionally, after prolonged stimulation, Teff cells can upregulate PD-1 and become dysfunctional. The combination of anti-CCR8-mediated Treg depletion with chemotherapy and PD-1 blockade can further enhance antitumor T cell immunity and cause inhibition of tumor growth.

[0138] Chemotherapy has the potential to trigger immunogenic cell death and result in enhanced T cell stimulation and activation. The present disclosure is at least partially based on the recognition that low-dose chemotherapy treatment can be combined with a Treg depleting agent, such as an anti-CCR8 antibody, and result in improved outcomes due to the ability of chemotherapy to induce immunogenic cell death. It is demonstrated that the combination of reduced Treg suppression and enhanced tumor antigen release during T cell stimulation acts together to result in stronger effector T cell activation and killing within the tumor microenvironment (see, e.g., Example 1).

[0139] In one aspect, provided herein is a method of treating cancer in a subject, the method comprising co-administering to the subject an effective amount of: (i) an anti-CCR8 antibody; (ii) a chemotherapeutic agent, and (iii) a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody); wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8 neutralizing antibody. In some embodiments, the chemotherapeutic agent is co-administered at a lower dose than in a standard of care chemotherapy regimen that does not include the anti-CCR8 antibody.

[0140] In another aspect, provided herein are methods of treating cancer in a subject, the methods comprising co-administering to the subject an effective amount of: (i) an anti-CCR8 antibody; (ii) a chemotherapeutic agent, and (iii) optionally a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody); wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity; wherein the anti-CCR8 antibody is optionally a CCR8 neutralizing antibody, and wherein the chemotherapeutic agent is administered at a lower dose than in a standard of care chemotherapy regimen that does not include the anti-CCR8 antibody.

[0141] Physicians (e.g., clinical oncologists) fully understand the administration of chemotherapeutic agents and regimens to subjects (such as human cancer patients). Standard of care (SOC) chemotherapy regimens are provided, for example, by medical societies such as the American Society of Clinical Oncology (ASCO) and the Oncology Nursing Society (ONS) (see, e.g., Neuss et al. Chemotherapy Administration, Guidelines, Safety, Standards, Pediatric Oncology. ONF 2017, 44(1), 31-41). In some embodiments of the methods provided herein, the chemotherapeutic agent is administered to the subject as part of an anti-CCR8 antibody combination therapy at a lower dose than the dose used in a standard of care chemotherapy regimen that does not include an anti-CCR8 antibody. In some embodiments, the dose of the chemotherapeutic agent administered is 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 50% or less, 40% or less, 30% or less, or 20% or less of the dose of the chemotherapeutic agent administered in a standard of care regimen that does not include an anti-CCR8 antibody.

[0142] In some embodiments of the methods provided herein, the co-administered chemotherapeutic agent is a single chemotherapeutic agent. In some embodiments, the co-administered chemotherapeutic agent is a plurality of chemotherapeutic agents. In some embodiments, the plurality of co-administered chemotherapeutic agents are two, three, four, or five chemotherapeutic agents.

[0143] In some embodiments of the methods provided herein, the dose of the chemotherapeutic agent administered is 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 50% or less, 40% or less, 30% or less, or 20% or less of the dose of the chemotherapeutic agent administered in a standard of care regimen that does not include an anti-CCR8 antibody.

[0144] In some embodiments, the cancer comprises a solid tumor.

[0145] In some embodiments, the cancer comprises tumor-infiltrating Treg cells that express CCR8. In some embodiments, CCR8 is expressed on the surface of Treg cells at less than 10,000 copies / cell (as determined by fluorescence-activated cell sorting (FACS) and / or flow cytometry). In some embodiments, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% of the tumor-infiltrating Tregs express less than 10,000 copies of CCR8 / cell on the cell surface.

[0146] In some embodiments, the cancer is selected from the group consisting of: breast cancer, colorectal cancer, head and neck cancer, lung cancer, ovarian cancer, gastric cancer, gastric adenocarcinoma, and thymoma. In some embodiments, the cancer is selected from the group consisting of: endometrial adenocarcinoma, colorectal cancer, ovarian cancer, vaginal squamous cell carcinoma, endometrial adenocarcinoma, colorectal cancer, cutaneous melanoma, pancreatic cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), uterine leiomyosarcoma, cholangiocarcinoma, adenoid cystic carcinoma, cervical cancer, anal cancer, esophagogastric junction (EGJ) adenocarcinoma, and gastric adenocarcinoma. In some embodiments, the cancer is selected from the group consisting of: head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer (NSCLC), gastric adenocarcinoma, EGJ adenocarcinoma, and colorectal cancer (CRC) (e.g., microsatellite stable (MSS) mCRC). In some embodiments, the cancer is selected from the group consisting of: breast cancer, pancreatic cancer, and lung cancer. In some embodiments, the breast cancer is selected from triple-negative breast cancer (TNBC), HR + / HER 2- breast cancer or HR + / HER2 低 breast cancer. In some embodiments, the pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC). In some embodiments, the cancer is metastatic.

[0147] In some embodiments, the cancer is a blood cancer that expresses CCR8. In some embodiments, the blood cancer is selected from the group consisting of: T-cell adult acute lymphoblastic leukemia, T-cell pediatric acute lymphoblastic leukemia, lymphoblastic lymphoma, acute lymphoblastic leukemia, cutaneous T-cell lymphoma (CTCL), T-cell acute lymphoblastic leukemia, adult T-cell leukemia / lymphoma, T-cell lymphoblastic leukemia / lymphoma, and anaplastic large cell lymphoma. In some embodiments, the blood cancer is CTCL.

[0148] In some embodiments, the subject is a human. In some embodiments, the subject has not received treatment. In some embodiments, the subject has received one or more courses of anti-cancer treatment, and optionally, the cancer has progressed during one or more courses of anti-cancer treatment. In some embodiments, the anti-cancer treatment during which the cancer has progressed is selected from the group consisting of surgery, radiotherapy, hormone therapy, targeted anti-cancer agents, chemotherapeutic agents, immunotherapy, and antibody-drug conjugates (ADCs). In some embodiments, the chemotherapeutic agents during which the cancer has progressed are selected from the group consisting of platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin. In some embodiments, the platinum complexes are selected from the group consisting of carboplatin, cisplatin, and oxaliplatin. In some embodiments, the taxane is paclitaxel, nab-paclitaxel, or docetaxel. In some embodiments, the taxane is docetaxel. In some embodiments, the immunotherapy during which the cancer has progressed comprises an anti-PD-1 antibody or an anti-PD-L1 antibody. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, tislelizumab, rivulizumab, btilimab, toripalimab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and serplimab. In some embodiments, the immunotherapy during which the cancer has progressed further comprises an anti-TIGIT antibody. In some embodiments, the anti-TIGIT antibody is selected from the group consisting of tirzepatide, vibostolimab, donanemab, AB308, AK127, BMS-986207, or eptinezumab.

[0149] In another aspect, the present disclosure provides an anti-CCR8 antibody for use in combination with a chemotherapeutic agent and a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) in a method of treating cancer, the method comprising co-administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the PD-1 inhibitor or the PD-L1 inhibitor (e.g., the anti-PD1 antibody or the anti-PD-L1 antibody), wherein the anti-CCR8 antibody has antibody-dependent cell cytotoxicity (ADCC) activity and / or complement-dependent cell cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8 neutralizing antibody.

[0150] In another aspect, the present disclosure provides an anti-CCR8 antibody for use in combination with a chemotherapeutic agent and optionally a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) in a method of treating cancer, the method comprising co-administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the PD-1 inhibitor or the PD-L1 inhibitor (e.g., the anti-PD1 antibody or the anti-PD-L1 antibody), wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody, and wherein the chemotherapeutic agent is administered at a lower dose than in a standard-of-care chemotherapy regimen that does not include the anti-CCR8 antibody.

[0151] Anti-CCR8 antibody

[0152] Anti-CCR8 antibodies useful in the methods provided herein generally have the ability to deplete target cells expressing CCR8, such as Tregs or cancer cells expressing CCR8. Such anti-CCR8 antibodies can include, but are not limited to, humanized antibodies, chimeric antibodies, murine antibodies, human antibodies, and antibodies comprising the heavy-chain CDRs and / or light-chain CDRs discussed herein. In some embodiments, an isolated antibody that binds CCR8 is used. In some embodiments, a monoclonal antibody that binds CCR8 is used. In some embodiments, the anti-CCR8 antibody is an antagonist anti-CCR8 antibody. In some embodiments, the anti-CCR8 antibody used in the methods provided herein inhibits the binding of CCR8 to CCL1. In some embodiments, co-administration of the anti-CCR8 antibody described herein reduces infiltrating Treg cells in the cancer of the subject. In some embodiments, co-administration of the anti-CCR8 antibody herein treats blood cancers expressing CCR8.

[0153] In some embodiments, the anti-CCR8 antibody useful in the methods provided herein is as described in International Patent Publication No. WO2021 / 163064.

[0154] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 12; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 13; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 14; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 15; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 16; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0155] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 24; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 25; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 26; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 27; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 28; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0156] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 36; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 37; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 38; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 39; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 40; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 41.

[0157] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 48; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 49; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 50; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 51; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 52; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 53.

[0158] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 60; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 61, 72, or 78; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 62, 73, or 79; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 63; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 64; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 65.

[0159] In some embodiments, the anti-CCR8 antibody comprises at least one, two, three, four, five, or six CDRs selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 84 or 100; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 85; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 86; (d) LCDR1 having an amino acid sequence selected from SEQ ID NO: 87; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 88; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 89.

[0160] In some embodiments, the anti-CCR8 antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the anti-CCR8 antibody comprises at least one heavy chain comprising a heavy chain variable region and at least a portion of a heavy chain constant region and at least one light chain comprising a light chain variable region and at least a portion of a light chain constant region. In some embodiments, the anti-CCR8 antibody comprises two heavy chains and two light chains, wherein each heavy chain comprises a heavy chain variable region and at least a portion of a heavy chain constant region, and wherein each light chain comprises a light chain variable region and at least a portion of a light chain constant region. As used herein, a single-chain Fv (scFv) or any other antibody comprising, for example, a single polypeptide chain containing all six CDRs (three heavy chain CDRs and three light chain CDRs) is considered to have a heavy chain and a light chain. In some embodiments, the heavy chain is the region of the anti-CCR8 antibody that contains the three heavy chain CDRs. In some embodiments, the light chain is the region of the anti-CCR8 antibody that contains the three light chain CDRs.

[0161] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 12; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 13; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 14; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 15; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 16; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 17.

[0162] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 24; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 25; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 26; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 27; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 28; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0163] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 36; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 37; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 38; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 39; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 40; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 41.

[0164] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 48; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 49; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 50; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 51; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 52; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 53.

[0165] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 60; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 61, 72, or 78; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 62, 73, or 79; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 63; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 64; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 65.

[0166] In some embodiments, the anti-CCR8 antibody comprises six CDRs, including: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 84 or 100; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 85; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 86; (d) LCDR1 having the amino acid sequence of SEQ ID NO: 87; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 88; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 89.

[0167] In some embodiments, the anti-CCR8 antibody comprises six CDRs as described above and binds to CCR8. In some embodiments, the anti-CCR8 antibody comprises six CDRs as described above, binds to CCR8 and inhibits the binding of CCR8 to CCL1. In some embodiments, the anti-CCR8 antibody comprises six CDRs as described above, binds to CCR8 and enhances the immune response of a subject, and / or increases the activation of T cells in the subject after administration of the antibody to the subject.

[0168] In some embodiments, an anti-CCR8 antibody that competes with the anti-CCR8 antibody described herein for binding to CCR8 is used. In some embodiments, an antibody that competes for binding with any of the antibodies described herein can be prepared and / or used.

[0169] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 12; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 13; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 14.

[0170] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 24; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 25; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 26.

[0171] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 36; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 37; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 38.

[0172] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 48; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 49; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 50.

[0173] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 60; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 61, 72, or 78; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 62, 73, or 79.

[0174] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 84 or 100; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 85; and (c) HCDR3 having the amino acid sequence of SEQ ID NO: 86.

[0175] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) LCDR1 having the amino acid sequence of SEQ ID NO: 15; (b) LCDR2 having the amino acid sequence of SEQ ID NO: 16; and (c) LCDR3 having the amino acid sequence of SEQ ID NO: 17.

[0176] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) an LCDR1 having the amino acid sequence of SEQ ID NO: 27; (b) an LCDR2 having the amino acid sequence of SEQ ID NO: 28; and (c) an LCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0177] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) an LCDR1 having the amino acid sequence of SEQ ID NO: 39; (b) an LCDR2 having the amino acid sequence of SEQ ID NO: 40; and (c) an LCDR3 having the amino acid sequence of SEQ ID NO: 41.

[0178] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) an LCDR1 having the amino acid sequence of SEQ ID NO: 51; (b) an LCDR2 having the amino acid sequence of SEQ ID NO: 52; and (c) an LCDR3 having the amino acid sequence of SEQ ID NO: 53.

[0179] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) an LCDR1 having the amino acid sequence of SEQ ID NO: 63; (b) an LCDR2 having the amino acid sequence of SEQ ID NO: 64; and (c) an LCDR3 having the amino acid sequence of SEQ ID NO: 65.

[0180] In some embodiments, the anti-CCR8 antibody comprises at least one, at least two, or all three VL CDR sequences selected from: (a) an LCDR1 having the amino acid sequence of SEQ ID NO: 87; (b) an LCDR2 having the amino acid sequence of SEQ ID NO: 88; and (c) an LCDR3 having the amino acid sequence of SEQ ID NO: 89.

[0181] In some embodiments, any one of the six CDRs described herein can be combined as a sub - part with any one of the other CDRs described herein, such that there are a total of six CDRs in the construct. Thus, in some embodiments, two CDRs from a first antibody (e.g., HCDR1 and HCDR2) can be combined with four CDRs from a second antibody (HCDR3, LCDR1, LCDR2, and LCDR3). In some embodiments, two or fewer residues in one or more CDRs can be substituted to obtain variants thereof. In some embodiments, two or fewer residues in 1, 2, 3, 4, 5, or 6 CDRs can be substituted.

[0182] In some embodiments, the anti - CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 12; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 13; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 14; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence of SEQ ID NO: 15; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 16; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 17.

[0183] In some embodiments, the anti - CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 24; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 25; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 26; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence of SEQ ID NO: 27; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 28; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0184] In some embodiments, the anti-CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence set forth in SEQ ID NO: 36; (b) HCDR2 having the amino acid sequence set forth in SEQ ID NO: 37; (c) HCDR3 having the amino acid sequence set forth in SEQ ID NO: 38; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence set forth in SEQ ID NO: 39; (e) LCDR2 having the amino acid sequence set forth in SEQ ID NO: 40; and (f) LCDR3 having the amino acid sequence set forth in SEQ ID NO: 41.

[0185] In some embodiments, the anti-CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence set forth in SEQ ID NO: 48; (b) HCDR2 having the amino acid sequence set forth in SEQ ID NO: 49; (c) HCDR3 having the amino acid sequence set forth in SEQ ID NO: 50; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence set forth in SEQ ID NO: 51; (e) LCDR2 having the amino acid sequence set forth in SEQ ID NO: 52; and (f) LCDR3 having the amino acid sequence set forth in SEQ ID NO: 53.

[0186] In some embodiments, the anti-CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence set forth in SEQ ID NO: 60; (b) HCDR2 having the amino acid sequence set forth in SEQ ID NO: 61, 72, or 78; (c) HCDR3 having the amino acid sequence set forth in SEQ ID NO: 62, 73, or 79; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence set forth in SEQ ID NO: 63; (e) LCDR2 having the amino acid sequence set forth in SEQ ID NO: 64; and (f) LCDR3 having the amino acid sequence set forth in SEQ ID NO: 65.

[0187] In some embodiments, the anti-CCR8 antibody comprises (I) a VH domain comprising at least one, at least two, or all three VH CDR sequences selected from: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 84 or 100; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 85; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 86; and (II) a VL domain comprising at least one, at least two, or all three VL CDR sequences selected from: (d) LCDR1 having the amino acid sequence of SEQ ID NO: 87; (e) LCDR2 having the amino acid sequence of SEQ ID NO: 88; and (f) LCDR3 having the amino acid sequence of SEQ ID NO: 89.

[0188] In some embodiments, 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 the amino acid sequence of SEQ ID NO: 68 or 74. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) in SEQ ID NO: 68 or 74 have been substituted, inserted, and / or deleted. In some embodiments, the substitutions, insertions, or deletions occur in regions outside of the CDRs (i.e., in the FRs). Optionally, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 68 or 74, including post-translational modifications of the sequence.

[0189] In some embodiments, the VH comprises: (a) HCDR1 having the amino acid sequence of SEQ ID NO: 60; (b) HCDR2 having the amino acid sequence of SEQ ID NO: 61, 72, or 78; (c) HCDR3 having the amino acid sequence of SEQ ID NO: 62, 73, or 79.

[0190] In some embodiments, 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 the amino acid sequence of SEQ ID NO: 92 or 96. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (such as conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind CCR8. In some embodiments, a total of 1 to 10 amino acids (such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 92 or 96 have been substituted, inserted and / or deleted. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). Optionally, the anti-CCR8 antibody comprises the VH sequence in SEQ ID NO: 92 or 96, including post-translational modifications of the sequence.

[0191] In some embodiments, the VH comprises: (a) an HCDR1 containing the amino acid sequence of SEQ ID NO: 84 or 100; (b) an HCDR2 containing the amino acid sequence of SEQ ID NO: 85; (c) an HCDR3 containing the amino acid sequence of SEQ ID NO: 86.

[0192] In some embodiments, there is provided an anti-CCR8 antibody, wherein the antibody comprises a light chain variable domain (VL) 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: 69 or 75. In some embodiments, the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (such as conservative substitutions), insertions or deletions relative to the reference sequence, but the anti-CCR8 antibody comprising the sequence retains the ability to bind CCR8. In some embodiments, a total of 1 to 10 amino acids (such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 69 or 75 have been substituted, inserted and / or deleted. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). Optionally, the anti-CCR8 antibody comprises the VL sequence in SEQ ID NO: 69 or 75, including post-translational modifications of the sequence.

[0193] In some embodiments, the VL comprises: (a) an LCDR1 comprising the amino acid sequence of SEQ ID NO: 63; (b) an LCDR2 comprising the amino acid sequence of SEQ ID NO: 64; and (c) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 65.

[0194] In some embodiments, an anti-CCR8 antibody is provided, wherein the antibody comprises a light chain variable domain (VL) 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: 93 or 97. In some embodiments, the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 93 or 97 have been substituted, inserted and / or deleted. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). Optionally, the anti-CCR8 antibody comprises the VL sequence of SEQ ID NO: 93 or 97, including post-translational modifications of the sequence.

[0195] In some embodiments, the VL comprises: (a) an LCDR1 comprising the amino acid sequence of SEQ ID NO: 87; (b) an LCDR2 comprising the amino acid sequence of SEQ ID NO: 88; and (c) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 89.

[0196] In some embodiments, 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 the amino acid sequence of SEQ ID NO: 68 or 74 and a light chain variable domain (VL) 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: 69 or 75. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, and the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) have been substituted, inserted and / or deleted in SEQ ID NO: 68 or 74. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) have been substituted, inserted and / or deleted in SEQ ID NO: 69 or 75. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). In some embodiments, the anti-CCR8 antibody comprises: (a) an HCDR1 containing the amino acid sequence of SEQ ID NO: 60; (b) an HCDR2 containing the amino acid sequence of SEQ ID NO: 61, 72 or 78; (c) an HCDR3 containing the amino acid sequence of SEQ ID NO: 62, 73 or 79; (d) an LCDR1 containing the amino acid sequence of SEQ IDNO: 63; (e) an LCDR2 containing the amino acid sequence of SEQ ID NO: 64; and (f) an LCDR3 containing the amino acid sequence of SEQ ID NO: 65.

[0197] In some embodiments, the anti-CCR8 antibody comprises the VH sequence of SEQ ID NO: 68 or 74, including post-translational modifications of one or both sequences, and comprises the VL sequence of SEQ ID NO: 69 or 75, including post-translational modifications of one or both sequences.

[0198] In some embodiments, 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 the amino acid sequence of SEQ ID NO: 92 or 96 and a light chain variable domain (VL) 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: 93 or 97. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, and the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 92 or 96 have been substituted, inserted and / or deleted. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 93 or 97 have been substituted, inserted and / or deleted. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). In some embodiments, the anti-CCR8 antibody comprises: (a) an HCDR1 containing the amino acid sequence of SEQ ID NO: 84 or 100; (b) an HCDR2 containing the amino acid sequence of SEQ ID NO: 85; (c) an HCDR3 containing the amino acid sequence of SEQ ID NO: 86; (d) an LCDR1 containing the amino acid sequence of SEQ ID NO: 87; (e) an LCDR2 containing the amino acid sequence of SEQ ID NO: 88; and (f) an LCDR3 containing the amino acid sequence of SEQ ID NO: 89.

[0199] In some embodiments, the anti-CCR8 antibody comprises the VH sequence in SEQ ID NO: 92 or 96, including post-translational modifications of one or both sequences, and comprises the VL sequence in SEQ ID NO: 93 or 97, including post-translational modifications of one or both sequences.

[0200] In some embodiments, the anti-CCR8 antibody comprises a VH as in any of the embodiments provided herein and a VL as in any of the embodiments provided herein. In some embodiments, the antibody comprises VH and VL sequences in SEQ ID NO: 68 or 74 and SEQ ID NO: 69 or 75, respectively, including post-translational modifications of those sequences. In some embodiments, the antibody comprises VH and VL sequences in SEQ ID NO: 92 or 96 and SEQ ID NO: 93 or 97, respectively, including post-translational modifications of those sequences.

[0201] In some embodiments, an anti-CCR8 antibody is used in the methods provided herein, wherein the antibody comprises a heavy chain (HC) 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 or 76. Optionally, the anti-CCR8 antibody comprises the HC sequence in SEQ ID NO: 70 or 76, including post-translational modifications.

[0202] In some embodiments, an anti-CCR8 antibody is used in the methods provided herein, wherein the antibody comprises an HC 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: 94 or 98. Optionally, the anti-CCR8 antibody comprises the HC sequence in SEQ ID NO: 94 or 98, including post-translational modifications.

[0203] In some embodiments, an anti-CCR8 antibody is used in the methods provided herein, wherein the antibody comprises a light chain (LC) 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: 71 or 77. Optionally, the anti-CCR8 antibody comprises the LC sequence in SEQ ID NO: 71 or 77, including post-translational modifications.

[0204] In some embodiments, an anti-CCR8 antibody is used in the methods provided herein, wherein the antibody comprises an LC 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 or 99. Optionally, the anti-CCR8 antibody comprises the LC sequence in SEQ ID NO: 95 or 99, including post-translational modifications.

[0205] In some embodiments, the anti-CCR8 antibody comprises an HC as in any of the embodiments provided herein and an LC as in any of the embodiments provided herein. In some embodiments, the antibody comprises HC and LC sequences in SEQ ID NO: 70 or 76 and SEQ ID NO: 71 or 77, respectively, including post-translational modifications of those sequences. In some embodiments, the antibody comprises HC and LC sequences in SEQ ID NO: 94 or 98 and SEQ ID NO: 95 or 99, respectively, including post-translational modifications of those sequences.

[0206] In some embodiments, an antibody that competes with the anti-CCR8 antibody described herein for binding to CCR8 is used in the methods provided herein. In some embodiments, the antibody competes with the anti-CCR8 antibody provided herein for binding to an epitope on CCR8.

[0207] In some embodiments, competition assays can be used to identify monoclonal antibodies that compete with the anti-CCR8 antibodies described herein (such as 1-K16, 1-K17, 6-B09, 7-B16, 13-E16, and / or 19-O07) for binding to CCR8. Competition assays can be used to determine whether two antibodies bind the same epitope by recognizing the same or spatially overlapping epitopes, or whether one antibody competitively inhibits the binding of another antibody to an antigen. In some embodiments, such competing antibodies bind the same epitope as the epitope bound by the antibodies described herein. Exemplary competition assays include, but are not limited to, conventional assays such as those provided in Harlow and Lane (1988) Antibodies: A Laboratory Manual, Chapter 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.). Detailed exemplary methods for mapping the epitopes bound by antibodies are provided in Morris (1996) “Epitope Mapping Protocols,” in Methods in Molecular Biology, Volume 66 (Humana Press, Totowa, N.J.). In some embodiments, two antibodies are said to bind the same epitope if each antibody blocks 50% or more of the binding of the other antibody. In some embodiments, the antibody that competes with the anti-CCR8 antibody described herein is a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, an antibody that competes with a chimeric, humanized, or human anti-CCR8 antibody as described herein is provided.

[0208] In addition, the present disclosure also includes variants of the antibodies disclosed above, such as variants of the 7-B16 antibody. For example, in some embodiments, the present disclosure includes an isolated antibody that binds to human CCR8, wherein the antibody comprises an HCDR3 that comprises SEQ ID NO: 86 or a variant of SEQ ID NO: 86 having 1, 2, or 3 mutations, and wherein the antibody binds to human CCR8 and has ADCC activity. In some embodiments, the mutations are substitutions (e.g., conservative or non-conservative substitutions), deletions, or insertions. In some embodiments, the 1, 2, or 3 mutations are located at at least one amino acid position among amino acid positions 1-4, 6, 7, or 12 of SEQ ID NO: 86. In some embodiments, the substitution is a conservative substitution. In some embodiments, the conservative substitution is located at amino acid positions 1, 4, or 12 of SEQ ID NO: 86. In some embodiments, the substitution is a non-conservative substitution. In some embodiments, the non-conservative substitution is located at amino acid position 7 of SEQ ID NO: 86. In some embodiments, the antibody comprises at least 2 substitutions in the HCDR3. In some embodiments, the at least 2 substitutions are located at at least one amino acid position among amino acid positions 1-4, 6, 7, or 12 of SEQ ID NO: 86. In some embodiments, the at least 2 substitutions are conservative substitutions. In some embodiments, at least one conservative substitution is located at amino acid positions 1, 4, or 12 of SEQ ID NO: 86. In some embodiments, the at least 2 substitutions are non-conservative substitutions. In some embodiments, at least one non-conservative substitution is located at amino acid position 7 of SEQ ID NO: 86. In some embodiments, when there are more than one substitution mutations, the mutations comprise conservative substitutions and non-conservative substitutions. In some embodiments, the present disclosure provides an isolated antibody that binds to human CCR8, wherein the antibody comprises an HCDR3 that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 86, and wherein the antibody binds to human CCR8 and has ADCC activity. In some embodiments, the HCDR3 comprises an amino acid sequence selected from any one of SEQ ID NO: 86 and SEQ ID NO: 104-119. In some embodiments, the antibody comprises an HCDR1 comprising SEQ ID NO: 84 or SEQ ID NO: 123. In some embodiments, the antibody comprises an HCDR2 comprising SEQ ID NO: 85 or SEQ ID NO: 124. In some embodiments, the antibody comprises an LCDR1 comprising SEQ ID NO: 87 or SEQ ID NO: 120.In some embodiments, the antibody comprises an LCDR2 comprising SEQ ID NO: 88 or SEQ ID NO: 121. In some embodiments, the antibody comprises an LCDR3 comprising SEQ ID NO: 89 or SEQ ID NO: 122. In some embodiments, the ADCC activity comprises an EC50 value of less than 200 ng / ml, 175 ng / ml, 150 ng / ml, 125 ng / ml, 100 ng / ml, 75 ng / ml, 50 ng / ml, 25 ng / ml, 20 ng / ml, 15 ng / ml, 10 ng / ml, 9 ng / ml, 8 ng / ml, 7 ng / ml, 6 ng / ml, 5 ng / ml, 4 ng / ml, 3 ng / ml, 2 ng / ml, or 1 ng / ml, as measured by a bioassay based on the ADCC reporter mechanism of action (MOA). In some embodiments, the ADCC activity is more potent than the 7-B16 antibody. In some embodiments, the ADCC activity is at least as potent as the 7-B16 antibody. In some embodiments, the antibody has a K for human CCR8. D equal to or lower than the 7-B16 antibody (e.g., as determined by kinetic exclusion assay (i.e., KinExA)). In some embodiments, the antibody has a cell surface K for human CCR8 DEqual to or less than the 7-B16 antibody (e.g., as determined by kinetic exclusion assay (i.e., KinExA)). In some embodiments, the antibody comprises at least one modification that enhances cell killing. In some embodiments, the enhanced cell killing is enhanced antibody-dependent cell cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC). In some embodiments, the at least one modification is afucosylation. In some embodiments, the at least one modification is one or more heavy chain constant region mutations at one or more positions selected from L234, L235, G236, S239, F243, H268, D270, R292, S298, Y300, V305, K326, A330, I332, E333, K334, and P396. In some embodiments, the one or more heavy chain constant region mutations are one or more mutations selected from S239D, S239M, F243L, H268D, D270E, R292P, S298A, Y300L, V305I, K326D, A330L, A330M, I332E, E333A, K334A, K334E, and P396L. In some embodiments, the one or more heavy chain constant region mutations are selected from: F243L / R292P / Y300L / V305I / P396L, S239D / I332E, S239D / I332E / A330L, S298A / E333A / K334A, L234Y / L235Q / G236W / S239M / H268D / D270E / S298A, and D270E / K326D / A330M / K334E. In some embodiments, the at least one modification is galactosylation. In some embodiments, the antibody binds to human CCR8 with an affinity (K D ), as determined by kinetic exclusion assay (i.e., KinExA), of less than 10 nM, or less than 5 nM, or less than 1 nM, or less than 500 pM, or less than 250 pM, or less than 100 pM, or less than 75 pM, or less than 50 pM, or less than 25 pM. In some embodiments, the antibody binds to human CCR8 with a cell surface affinity (K D ), as determined by, for example, kinetic exclusion assay (i.e., KinExA), of less than 10 nM, or less than 5 nM, or less than 1 nM, or less than 500 pM, or less than 250 pM, or less than 100 pM, or less than 75 pM, or less than 50 pM, or less than 25 pM. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a human or humanized antibody. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is an IgG1 or IgG3 antibody. Such variants can be used in methods for treating cancer (including both blood cancers and solid tumors).

[0209] In some embodiments, an antibody that binds to any one or more of the epitopes bound by the antibodies described herein can be used in the methods provided herein. In some embodiments, antibodies that bind to and overlap the epitopes bound by the antibodies of the invention are described. In some embodiments, an antibody that competes with at least one of the antibodies described herein is used. In some embodiments, an antibody that competes with at least two of the antibodies described herein is used. In some embodiments, an antibody that competes with at least three of the antibodies described herein is used. In some embodiments, the entire epitope is bound and / or blocked by the competing antibody. In some embodiments, a portion of the epitope is bound and / or blocked by the competing antibody. In some embodiments, the paratope of the competing antibody binds to at least a portion of the epitope of the antibody provided herein. In some embodiments, the paratope of the competing antibody binds to the target, and a different portion of the structure of the competing antibody blocks at least a portion of the epitope of the antibody provided herein.

[0210] Exemplary chimeric anti-CCR8 antibody

[0211] In some embodiments, the anti-CCR8 antibodies useful in the methods described herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In one example, a chimeric antibody comprises 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-switch" antibody in which the class or subclass has been changed from that of the parental antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0212] Non-limiting exemplary chimeric antibodies include chimeric antibodies comprising a heavy chain variable region and / or a light chain variable region of an antibody selected from, for example, antibodies 1-K16, 1-K17, 6-B09, 7-B16, 13-E16, and 19-O07 disclosed herein. Additional non-limiting exemplary chimeric antibodies include chimeric antibodies comprising heavy chain CDR1, CDR2, and CDR3 and / or light chain CDR1, CDR2, and CDR3 of an antibody selected from antibodies 1-K16, 1-K17, 6-B09, 7-B16, 13-E16, and 19-O07 as disclosed herein. In some embodiments, the chimeric anti-CCR8 antibody comprises the variable regions described above and binds CCR8. In some embodiments, the chimeric anti-CCR8 antibody comprises the variable regions described above, binds CCR8 and inhibits the binding of CCR8 to CCL1. In some embodiments, the anti-CCR8 antibody comprises the variable regions described above, binds CCR8 and enhances the immune response of a subject, and / or increases the activation of T cells in a subject after administration of the antibody to the subject. In some embodiments, administration of the anti-CCR8 antibody described herein stimulates the activity of immune cells in a subject, reduces the downregulation of immune cells, or increases the T cell response.

[0213] In some embodiments, the chimeric antibodies described herein comprise one or more human constant regions. In some embodiments, the human heavy chain constant region has an isotype selected from IgA, IgG, IgD, and IgE. In some embodiments, the human light chain constant region has an isotype selected from κ and λ. In some embodiments, the chimeric antibodies described herein comprise a human IgG constant region. In some embodiments, the chimeric antibodies described herein comprise a human IgG4 heavy chain constant region. In some embodiments, the chimeric antibodies described herein comprise a human IgG4 constant region and a human κ light chain.

[0214] As described above, whether effector function is desirable may depend on the particular therapeutic method intended for the antibody. Thus, in some embodiments, when effector function is desired, a chimeric anti-CCR8 antibody comprising a human IgG1 heavy chain constant region or a human IgG3 heavy chain constant region is selected. In some embodiments, when effector function is not desired, a chimeric anti-CCR8 antibody comprising a human IgG4 or IgG2 heavy chain constant region is selected. In some embodiments, enhanced effector function is desired.

[0215] Exemplary humanized anti-CCR8 antibodies

[0216] In some embodiments, a humanized antibody that binds CCR8 can be used in the methods provided herein. Humanized antibodies can be used as therapeutic molecules because humanized antibodies reduce or eliminate the human immune response compared to non-human antibodies, which can lead to an immune response to the antibody therapeutic (such as a human anti-mouse antibody (HAMA) response) and reduced therapeutic agent efficacy.

[0217] In some embodiments, the chimeric antibody is a humanized antibody. Generally, non-human antibodies are humanized to reduce their immunogenicity in humans while retaining the specificity and affinity of the parental non-human antibody. Typically, a humanized antibody comprises one or more variable domains in which the CDRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from a human antibody sequence. A humanized antibody optionally also will comprise at least a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody are replaced with the corresponding residues of a non-human antibody (e.g., the antibody from which the CDR residues are derived), for example to restore or improve antibody specificity or affinity.

[0218] Reviews of humanized antibodies and methods of making them are provided, for example, in Almagro and Fransson, (2008) Front. Biosci. 13: 1619-1633, and are further described, for example, in Riechmann et al., (1988) Nature 332:323-329; Queen et al., (1989) Proc. Natl Acad. Sci. USA 86: 10029-10033; U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., (2005) Methods 36:25-34; Padlan, (1991) Mol. Immunol. 28:489-498 (describing "reshaping"); Dall'Acqua et al., (2005) Methods 36:43-60 (describing "FR shuffling"); and Osbourn et al., (2005) Methods 36:61-68 and Klimka et al., (2000) Br. J. Cancer, 83:252-260 (describing a "directed selection" method for FR shuffling).

[0219] 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. (1993) J. Immunol. 151:2296); 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. (1992) Proc. Natl. Acad. Sci. USA, 89:4285; and Presta et al. (1993) J. Immunol, 151:2623); human mature (somatic mutation) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, (2008) Front. Biosci. 13:1619-1633); and framework regions derived from screening FR libraries (see, e.g., Baca et al., (1997) J. Biol. Chem. 272: 10678-10684 and Rosok et al., (1996) J. Biol. Chem. 271:22611-22618).

[0220] In some embodiments, a humanized 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 the amino acid sequence of SEQ ID NO: 68 or 74 and a light chain variable domain (VL) 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: 69 or 75. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, and the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) have been substituted, inserted and / or deleted in SEQ ID NO: 68 or 74. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) have been substituted, inserted and / or deleted in SEQ ID NO: 69 or 75. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). In some embodiments, the anti-CCR8 antibody comprises: (a) an HCDR1 containing the amino acid sequence of SEQ ID NO: 60; (b) an HCDR2 containing the amino acid sequence of SEQ ID NO: 61, 72 or 78; (c) an HCDR3 containing the amino acid sequence of SEQ ID NO: 62, 73 or 79; (d) an LCDR1 containing the amino acid sequence of SEQ ID NO: 63; (e) an LCDR2 containing the amino acid sequence of SEQ ID NO: 64; and (f) an LCDR3 containing the amino acid sequence of SEQ ID NO: 65. In some embodiments, the antibody has ADCC activity.In some embodiments, the ADCC activity comprises an EC50 value of less than 200 ng / ml, 175 ng / ml, 150 ng / ml, 125 ng / ml, 100 ng / ml, 75 ng / ml, 50 ng / ml, 25 ng / ml, 20 ng / ml, 15 ng / ml, 10 ng / ml, 9 ng / ml, 8 ng / ml, 7 ng / ml, 6 ng / ml, 5 ng / ml, 4 ng / ml, 3 ng / ml, 2 ng / ml, or 1 ng / ml, as measured by a bioassay based on the ADCC reporter mechanism of action (MOA). In some embodiments, the ADCC activity is more potent than the 7-B16 antibody. In some embodiments, the ADCC activity is at least as potent as the 7-B16 antibody. In some embodiments, the K of the antibody for human CCR8. D is equal to or lower than that of the 7-B16 antibody (e.g., as determined by kinetic exclusion assay (i.e., KinExA)). In some embodiments, the on-cell K of the antibody for human CCR8 D is equal to or lower than that of the 7-B16 antibody (e.g., as determined by kinetic exclusion assay (i.e., KinExA)).

[0221] In some embodiments, the humanized anti-CCR8 antibody comprises a VH sequence of SEQ ID NO: 68 or 74, including post-translational modifications of one or both sequences, and comprises a VL sequence of SEQ ID NO: 69 or 75, including post-translational modifications of one or both sequences.

[0222] In some embodiments, a humanized 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 the amino acid sequence of SEQ ID NO: 92 or 96 and a light chain variable domain (VL) 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: 93 or 97. In some embodiments, the VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, and the VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 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 CCR8. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 92 or 96 have been substituted, inserted and / or deleted. In some embodiments, a total of 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids) in SEQ ID NO: 93 or 97 have been substituted, inserted and / or deleted. In some embodiments, the substitutions, insertions or deletions occur in regions outside the CDRs (i.e., in the FRs). In some embodiments, the anti-CCR8 antibody comprises: (a) an HCDR1 containing the amino acid sequence of SEQ ID NO: 84 or 100; (b) an HCDR2 containing the amino acid sequence of SEQ ID NO: 85; (c) an HCDR3 containing the amino acid sequence of SEQ ID NO: 86; (d) an LCDR1 containing the amino acid sequence of SEQ ID NO: 87; (e) an LCDR2 containing the amino acid sequence of SEQ ID NO: 88; and (f) an LCDR3 containing the amino acid sequence of SEQ ID NO: 89.

[0223] In some embodiments, a humanized anti-CCR8 antibody comprises the VH sequence in SEQ ID NO: 92 or 96, including post-translational modifications of one or both sequences, and comprises the VL sequence in SEQ ID NO: 93 or 97, including post-translational modifications of one or both sequences.

[0224] Exemplary humanized anti-CCR8 antibodies include antibodies that compete with the antibodies or fragments thereof described herein for binding to CCR8. Thus, in some embodiments, provided are humanized anti-CCR8 antibodies that compete with an antibody or fragment thereof selected from antibody 1-K16, 1-K17, 6-B09, 7-B16, 13-E16, and 19-O07 for binding to CCR8. In some embodiments, the humanized anti-CCR8 antibody competes with the antibodies described herein for binding to CCR8 and inhibits the binding of CCR8 to CCL1. In some embodiments, the humanized anti-CCR8 antibody competes with the antibodies described herein for binding to CCR8.

[0225] Exemplary human anti-CCR8 antibodies

[0226] In some embodiments, the anti-CCR8 antibodies used in the methods provided herein are human antibodies. A variety of techniques known in the art can be used to generate human antibodies. Human antibodies are generally described in van Dijk and van de Winkel, (2001) Curr. Opin. Pharmacol. 5:368-374 and Lonberg, (2008) Curr. Opin. Immunol. 20:450-459. In some embodiments, the human antibodies are not naturally occurring antibodies. In some embodiments, the human antibodies are monoclonal antibodies; thus, in some embodiments, each human antibody in a group can bind to the same epitope on the antigen.

[0227] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies having human variable regions in response to antigen challenge. Such animals typically contain all or part of the human immunoglobulin locus, which replaces the endogenous immunoglobulin locus or is present extrachromosomally or randomly integrated into the animal's chromosomes. 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, (2005) Nat. Biotech. 23:1117-1125. See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe the XENOMOUSE ™ technology; U.S. Patent No. 5,770,429, which describes the HUMAB ® technology; U.S. Patent No. 7,041,870, which describes the K-M MOUSE ® technology; and U.S. Patent Application Publication No. US 2007 / 0061900, which describes the VELOCIMOUSE ® technology. The human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining them with different human constant regions.

[0228] Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines useful for the production of human monoclonal antibodies have been described. (See, e.g., Kozbor (1984) J. Immunol, 133:3001; Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., (1991) J. Immunol., 147:86). Human antibodies produced by human B-cell hybridoma technology are also described in Li et al., (2006) Proc. Natl. Acad. Sci. USA, 103:3557-3562. Additional methods include, for example, those described in U.S. Patent No. 7,189,826 (describing the production of human IgM monoclonal antibodies from hybridoma cell lines) and Ni, (2006) Xiandai Mianyixue, 26(4):265-268 (describing human-human hybridomas). Human hybridoma technology (trioma technology) is also described in Vollmers and Brandlein, (2005) Histology and Histopathology, 20(3):927-937 (2005) and Vollmers and Brandlein, (2005) Methods and Findings in Experimental and Clinical Pharmacology, 27(3): 185-191.

[0229] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from a phage display library of human origin. Such variable domain sequences can then be combined with the desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0230] Antibodies can be isolated by screening combinatorial libraries for antibodies having one or more desired activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies having desired binding characteristics. Such methods are reviewed, for example, in Hoogenboom et al., in Methods in Molecular Biology 178: 1-37 (edited by O'Brien et al., Human Press, Totowa, NJ, 2001), and are further described, for example, in McCafferty et al., (1990) Nature 348:552-554; Clackson et al., (1991) Nature 352: 624-628; Marks et al., (1992) J. Mol. Biol 222: 581-597; Marks and Bradbury, in Methods in Molecular Biology 248: 161-175 (edited by Lo, Human Press, Totowa, NJ, 2003); Sidhu et al., (2004) J. Mol. Biol. 338(2): 299-310; Lee et al., (2004) J. Mol. Biol. 340(5):1073-1093; Fellouse, (2004) Proc. Natl. Acad. Sci. USA 101(34): 12467-12472; and Lee et al., (2004) J. Immunol. Methods 284(1-2): 119-132 and PCT Publication WO 99 / 10494.

[0231] In certain phage display methods, V H and V LGenomic libraries are cloned individually by polymerase chain reaction (PCR) and randomly recombined in a phage library, and antigen-binding phages in the library can then be screened, as described by Winter et al., (1994) Ann.Rev. Immunol., 12:433-455. Phages typically display antibody fragments, either as single-chain Fv (scFv) fragments or as Fab fragments. Libraries from immune sources provide high-affinity antibodies against the immunogen without the need to construct hybridomas. Alternatively, a naive library can be cloned (e.g., from humans) to provide a single source of antibodies against a wide range of non-self and self antigens without any immunization, as described by Griffiths et al., (1993) EMBO J 12:725-734. Finally, naive libraries can also be prepared synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences to encode the highly variable CDR3 region and achieve in vitro rearrangement, as described in Hoogenboom and Winter (1992), J. Mol.Biol, 227:381-388. Patent publications describing human antibody phage libraries include, for example: U.S. Patent No. 5,750,373 and U.S. Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0232] In some embodiments, the human anti-CCR8 antibody binds to CCR8 and inhibits the binding of CCR8 to CCL1.

[0233] Exemplary human anti-CCR8 antibodies also include antibodies that compete with the human antibodies or fragments thereof described herein for binding to CCR8. Thus, in some embodiments, there are provided human anti-CCR8 antibodies that compete with an antibody or fragment thereof selected from antibody 1-K16, 1-K17, 6-B09, 7-B16, 13-E16, and 19-O07 for binding to CCR8. In some embodiments, the human anti-CCR8 antibody competes with the antibodies described herein for binding to CCR8 and inhibits the binding of CCR8 to CCL1.

[0234] In some embodiments, there are provided chimeric human anti-CCR8 antibodies, wherein the antibody comprises a variable region from a human antibody that binds CCR8 and a constant region from a different human antibody. In some embodiments, there are provided chimeric human anti-CCR8 antibodies, wherein the antibody comprises the CDRs from a human antibody that binds CCR8 and a framework region from a different human antibody. In some embodiments, the antibody is not a naturally occurring human antibody.

[0235] In some embodiments, the human anti-CCR8 antibody comprises one or more human constant regions. In some embodiments, the human heavy chain constant region has an isotype selected from IgA, IgG, IgD, and IgE. In some embodiments, the human light chain constant region has an isotype selected from κ and λ. In some embodiments, the human antibodies described herein comprise a human IgG constant region. In some embodiments, the human antibodies described herein comprise a human IgG4 heavy chain constant region. In some embodiments, the human antibodies described herein comprise a human IgG4 constant region and a human κ light chain.

[0236] In some embodiments, when effector function is desired, a human anti-CCR8 antibody comprising a human IgG1 heavy chain constant region or a human IgG3 heavy chain constant region is selected. In some embodiments, when effector function is not desired, a human anti-CCR8 antibody comprising a human IgG4 or IgG2 heavy chain constant region is selected.

[0237] As used herein, the term "human antibody" refers to a genus of possible sequences of an antibody construct, rather than the source of the antibody.

[0238] Exemplary anti-CCR8 antibody constant region and Fc region

[0239] In some embodiments, the antibodies described herein comprise one or more human constant regions. In some embodiments, the human heavy chain constant region has an isotype selected from IgA, IgG, IgD, and IgE. In some embodiments, the antibodies described herein comprise a human IgG constant region. In some embodiments, when effector function is desired, an anti-CCR8 antibody comprising a human IgG1 heavy chain constant region or a human IgG3 heavy chain constant region is selected. In some embodiments, the human light chain constant region has an isotype selected from κ and λ. In some embodiments, the antibodies described herein comprise a human IgG1 heavy chain constant region. In some embodiments, the antibodies described herein comprise a human IgG1 constant region and a human κ light chain.

[0240] In some embodiments, the fusion proteins described herein comprise one or more human Fc regions. In some embodiments, the Fc region has an isotype selected from IgA, IgG, IgD, and IgE. In some embodiments, the fusion proteins described herein comprise a human Fc region. In some embodiments, when effector function is desired, a fusion protein comprising a human IgG1 Fc region or a human IgG3 Fc region is selected.

[0241] Throughout this specification and the claims, unless otherwise specified or known to those of skill in the art, the numbering of residues in immunoglobulin heavy chains is according to the EU index numbering as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), which is hereby incorporated by reference in its entirety. "EU index in Kabat" refers to the residue numbering of human IgG1 EU antibody.

[0242] As described above, whether effector function is desirable may depend on the particular therapeutic method intended for the antibody. Thus, in some embodiments, when effector function is desired, an anti-CCR8 antibody comprising a human IgG1 heavy chain constant region or a human IgG3 heavy chain constant region is selected.

[0243] In some embodiments, the antibody comprises a variant Fc region having at least one amino acid substitution compared to the Fc region of the wild-type IgG Fc region. In some embodiments, the variant Fc region has two or more amino acid substitutions compared to the wild-type Fc region. In some embodiments, the variant Fc region has three or more amino acid substitutions compared to the wild-type Fc region. In some embodiments, the variant Fc region has at least one, two, or three or more of the Fc region amino acid substitutions described herein. In some embodiments, the variant Fc region herein will have at least about 80% homology with the native sequence Fc region and / or with the Fc region of the parental polypeptide. In some embodiments, the variant Fc region herein will have at least about 90% homology with the native sequence Fc region and / or with the Fc region of the parental polypeptide. In some embodiments, the variant Fc region herein will have at least about 95% homology with the native sequence Fc region and / or with the Fc region of the parental polypeptide. In some embodiments, the heavy chain constant region or Fc region lacks a C-terminal lysine (K) residue. In some such embodiments, the heavy chain constant region or Fc region may be referred to as "desK". In some embodiments, the heavy chain constant region or Fc region lacking a C-terminal lysine is IgG, such as IgG1, IgG2, IgG3, or IgG4.

[0244] In some embodiments, the antibodies or fusion proteins provided herein are modified to increase or decrease the degree to which the antibody is glycosylated. Addition or deletion of antibody glycosylation sites can be conveniently achieved by altering the amino acid sequence so as to create or remove one or more glycosylation sites.

[0245] The carbohydrates linked to the Fc region can be altered. Native antibodies produced by mammalian cells typically contain branched biantennary oligosaccharides, which are generally linked by N-linkage to Asn297 of the CH2 domain of the Fc region. See, e.g., Wright et al., TIBTECH 15:26-32 (1997). The oligosaccharides can include various carbohydrates such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose linked to GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, the oligosaccharides in an antibody or fusion protein can be modified to generate antibody variants with certain improved properties.

[0246] In some embodiments, antibody or fusion protein variants are provided that have carbohydrate structures lacking fucose (i.e., are afucosylated) that are (directly or indirectly) linked to the Fc region. For example, the amount of fucose in such variants can be from 1% to 80%, from 1% to 65%, from 5% to 65%, or from 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose in the sugar chains at Asn297 relative to the sum of all sugar structures (e.g., complex, hybrid, and high mannose structures) linked to Asn297, as measured by MALDI-TOF mass spectrometry, as described, for example, in WO 2008 / 077546. Asn297 refers to the asparagine residue located at approximately position 297 in the Fc region (EU numbering of Fc region residues); however, due to minor sequence variations in the antibody, Asn297 can also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. Such fucosylation variants can have improved ADCC function. See, for example, U.S. Patent Publication No. US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "defucosylated" or "fucose-deficient" antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US 2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al. J. Mol.Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech., Bioeng. 87:614 (2004).Examples of cell lines capable of producing afucosylated antibodies include the Lec13 CHO cell line lacking protein fucosylation (Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986); US Patent Application No. US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., particularly in Example 11), and knockout cell lines such as the α-1,6-fucosyltransferase gene FUT8 knockout CHO cell line (see, e.g., Yamane-Ohnuki et al., Biotech., Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107).

[0247] Antibody variants also have bisected oligosaccharides, e.g., where the biantennary oligosaccharide linked to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants can have reduced fucosylation and / or improved ADCC function. Examples of such variants are described, e.g., in WO 2003 / 011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); and US2005 / 0123546 (Umana et al.). Variants having at least one galactose residue in the oligosaccharide linked to the Fc region are also provided. Such variants can have improved CDC function. Such variants are described, e.g., in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0248] Antibodies or Fc region variants have Fc mutations that increase ADCC activity. In some embodiments, the antibody or Fc region variant comprises one or more mutations that enhance FcγRIIIa binding and / or reduce FcγRIIIb binding. Such mutations can be made at one or more amino acid positions selected from L234, L235, G236, S239, F243, H268, D270, R292, S298, Y300, V305, K326, A330, I332, E333, K334, and P396. Non-limiting exemplary such mutations include L234Y, L235Q, G236W, S239D, S239M, F243L, H268D, D270E, R292P, S298A, Y300L, V305I, K326D, A330L, A330M, I332E, E333A, K334A, K334E, and P396L. In some embodiments, the antibody or Fc region variant comprises the mutations F243L / R292P / Y300L / V305I / P396L. See, e.g., Stavenhagen et al., 2007, Cancer Res. 67:8882-8890. In some embodiments, the antibody or Fc region variant comprises the mutations S239D / I332E or S239D / I332E / A330L. See, e.g., Lazar et al., 2006, PNAS USA, 103: 4005-4010. In some embodiments, the antibody or Fc region variant comprises the mutations S298A / E333A / K334A. See, e.g., Shields et al., 2001, J. Biol. Chem., 276: 6591-6604. In some embodiments, the antibody or Fc region variant comprises the mutations L234Y / L235Q / G236W / S239M / H268D / D270E / S298A or the mutations D270E / K326D / A330M / K334E, or one heavy chain constant region or Fc comprises the mutations L234Y / L235Q / G236W / S239M / H268D / D270E / S298A and the other heavy chain constant region or Fc comprises the mutations D270E / K326D / A330M / K334E. See, e.g., Mimoto et al., 2013, MAbs, 5:229-236.

[0249] Antibodies and Fc region variants also have an amino-terminal leader extension. For example, one or more amino acid residues of the amino-terminal leader sequence are present at the amino terminus of any one or more of the heavy or light chains of the antibody. Exemplary amino-terminal leader extensions comprise or consist of three amino acid residues VHS present on one or both light chains of the antibody variant.

[0250] The in vivo or serum half-life of a human FcRn high affinity binding polypeptide can be determined in, for example, transgenic mice, humans, or non-human primates to which a polypeptide having a variant Fc region is administered. See also, e.g., Petkova et al., International Immunology 18(12):1759-1769 (2006).

[0251] In some embodiments, an antibody or Fc region variant mediates ADCC more effectively than the parental antibody in the presence of human effector cells. In some embodiments, the antibody or Fc region variant is substantially more effective at mediating ADCC in vitro when the amounts of the polypeptide variant and the parental antibody or Fc region used in the assay are substantially the same. In some embodiments, the antibody or Fc region variant is substantially more effective at mediating ADCC in vivo when the amounts of the polypeptide variant and the parental antibody or Fc region used in the assay are substantially the same. Generally, such variants will be identified using an in vitro ADCC assay as disclosed herein, but other assays or methods for determining ADCC activity, e.g., in an animal model, are also contemplated.

[0252] Additional anti-CCR8 antibodies

[0253] In some embodiments, anti-CCR8 antibodies that can be used in the methods provided herein can include BMS-986340 (Bristol Myers Squibb), LM-108 (LaNova Medicines), S-531011 (Shionogi), FPA157 (Five Prime, Amgen), IPG-7236 (Immunophage Biomedical), ICP-B05 (InnoCare PharmaTech), SRF-114 (Surface Oncology), HBM1022 (Harbour BioMed), HFB1011 (HiFiBio), BAY-3375968 (Bayer), IO-1 (Oncurious), ZL-1218 (Zai Lab), GB2101 (Genor), or PSB-114 (SoundBiologics).

[0254] In some embodiments, the anti-CCR8 antibodies that can be used in the methods provided herein are as described in WO2022078277, WO2022081718, WO2022000443, WO2022042690, or WO2022003156.

[0255] In some embodiments, the anti-CCR8 antibodies useful in the methods provided herein can be obtained from hybridomas having ATCC accession numbers PTA-6940, PTA-6938, or PTA-6939.

[0256] In some embodiments, the anti-CCR8 antibody useful in the methods provided herein is the HBM1022 antibody as disclosed in Lu et al. HBM1022, a novel anti-CCR8 antibody, depletes tumor-infiltrating regulatory T cells by enhanced ADCC activity and mediates effective anti-tumor activity with Keytruda. Journal for ImmunoTherapy of Cancer 2020;8:doi: 10.1136 / jitc-2020-SITC2020.0711

[0257] In some embodiments, the anti-CCR8 antibody useful in the methods provided herein is the FPA157 antibody as disclosed in Rankin A, Naik E861 Development of FPA157, an anti-CCR8 depleting antibody engineered to preferentially eliminate tumor-infiltrating T regulatory cells. Journal for ImmunoTherapy of Cancer 2020; 8:doi:10.1136 / jitc-2020-SITC2020.0861

[0258] In some embodiments, the anti-CCR8 antibody useful in the methods provided herein is the SRF114 antibody as disclosed in Lake A, Warren M, Das S, et al726. SRF114 is a fully human CCR8-selective IgG1 antibody that induces tumor Treg disruption by ADCC. Journal for ImmunoTherapy of Cancer 2020; 8:doi: 10.1136 / jitc-2020-SITC2020.0726

[0259] In some embodiments, the anti-CCR8 that can be used in the methods provided herein is the anti-CCR8 hlgG1 non-fucosylated BMS-986340 as disclosed in the following documents: Lan, Ruth et al. “Highly selective anti-CCR8 antibody-mediated depletion of regulatory T cells leads to potent antitumor activity alone and in combination with anti-PD-1 in preclinical models.” (2020): 6694-6694 and Bayati F, Mohammadi M, Valadi M, Jamshidi S, Foma AM, Sharif-Paghaleh E. The Therapeutic Potential of Regulatory T Cells: Challenges and Opportunities. Front Immunol. 2021;11:585819. Published January 15, 2021. doi: 10.3389 / fimmu.2020.585819

[0260] In some embodiments, the anti-CCR8 antibodies that can be used in the methods provided herein are nanobodies as disclosed in the following reference: Van Damme H, Dombrecht B, Kiss M, Roose H, Allen E, VanOvermeire E, Kancheva D, Martens L, Murgaski A, Bardet PMR, Blancke G, JansM, Bolli E, Martins MS, Elkrim Y, Dooley J, Boon L, Schwarze JK, Tacke F, Movahedi K, Vandamme N, Neyns B, Ocak S, Scheyltjens I, Vereecke L, Nana FA, Merchiers P, Laoui D, Van Ginderachter JA. Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy. J Immunother Cancer. February 2021;9(2):e001749. doi: 10.1136 / jitc-2020-001749. PMID: 33589525; PMCID: PMC7887378.

[0261] PD-1 inhibitor or PD-L1 inhibitor

[0262] The PD-1 inhibitor or PD-L1 inhibitor used in the methods provided herein can be a small molecule inhibitor or an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0263] Exemplary anti-PD-1 antibodies or anti-PD-L1 antibodies that can be co-administered in the methods provided herein include, for example, pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, toripalimab, reflimab, bintrafusp alfa, tislelizumab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and sepacatlizumab. In some embodiments, the anti-PD-1 antibody is sepacatlizumab.

[0264] Additional illustrative anti-PD-1 antibodies or anti-PD-L1 antibodies that can be co-administered in the methods provided herein include pembrolizumab, nivolumab, cemiplimab, pidilizumab, AMP-224, MEDI0680 (AMP-514), spartalizumab, atezolizumab, avelumab, durvalumab, BMS-936559, cosibelimab (CK-301), sasanlimab (PF-06801591), tislelizumab (BGB-A317), GLS-010 (WBP-3055), AK-103 (HX-008), AK-105, CS-1003, HLX-10, rivulizumab (MGA-012), BI-754091, batrilizumab (AGEN-2034), AMG-404, toripalimab (JS-001), cetrelimab (JNJ-63723283), genolimab (CBT-501), LZM-009, palovelimab (BCD-100), lodavalimab (LY-3300054), SHR-1201, camrelizumab (SHR-1210), Sym-021, budigalimab (ABBV-181), PD1-PIK, BAT-1306, avelumab (MSB0010718C), CX-072, CBT-502, dostarlimab (TSR-042), MSB-2311, JTX-4014, BGB-A333, SHR-1316, CS-1001 (WBP-3155), envafolimab (KN-035), sintilimab (IBI-308), HLX-20, KL-A167, STI-A1014, STI-A1015 (IMC-001), BCD-135, FAZ-053, TQB-2450, MDX1105-01, GS-4224, GS-4416, INCB086550, MAX10181, sepapalimab (AB122), spartalizumab (PDR-001), and compounds disclosed in WO2018195321, WO2020014643, WO2019160882, or WO2018195321,and multi-specific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-013 (PD-1 / LAG-3), FS-118 (LAG-3 / PD-L1), RO-7247669 (PD-1 / LAG-3), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), RO-7121661 (PD-1 / TIM-3), RG7769 (PD-1 / TIM-3), TAK-252 (PD-1 / OX40L), XmAb-20717 (PD-1 / CTLA4), AK-104 (CTLA4 / PD-1), FS-118 (LAG-3 / PD-L1), FPT-155 (CTLA4 / PD-L1 / CD28), GEN-1046 (PD-L1 / 4-1BB), bintrafusp α (M7824; PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM3 / PDL1) and INBRX-105 (4-1BB / PDL1).

[0265] In some embodiments, the anti-PD-1 antibody is selected from cemiplimab (AB122, GLS-010, WBP-3055), pembrolizumab (KEYTRUDA ® , MK-3475, SCH900475), nivolumab (OPDIVO ® , BMS-936558, MDX-1106), cemiplimab (LIBTAYO ® ; cemiplimab-rwlc, REGN-2810), pidilizumab (CT-011), AMG-404, MEDI0680 (AMP-514), spartalizumab (PDR001), tislelizumab (BGB-A317), toripalimab (JS-001), genolimab (CBT-501, APL-501, GB 226), camrelizumab (SHR-1210), sintilimab (TYVYT ®; IBI-308), dostarlimab (TSR-042, WBP-285), zalifrelimab (PF-06801591), cilifrelimab (JNJ-63723283), surufatinib (HLX-10), rivafolimab (MGA-012), basticlimab (AGEN-2034), palovelimab (BCD-100), buglizumab (ABBV-181), voprelizumab (JTX-4014), AK-103 (HX-008), AK-105, CS-1003, BI-754091, LZM-009, Sym-021, BAT-1306, PD1-PIK, and the bispecific inhibitor teprotumumab (MGD013; PD-1 / LAG-3), RG-6139 (RO-7247669 PD-1 / LAG-3), FS-118 (LAG-3 / PD-L1), RO-7121661 (PD-1 / TIM-3), RG7769 (PD-1 / TIM-3), TAK-252 (PD-1 / OX40L), PF-06936308 (PD-1 / CTLA4), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), XmAb-20717 (PD-1 / CTLA4), AK-104 (CTLA4 / PD-1), and MEDI-5752 (CTLA4 / PD-1).

[0266] In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of: atezolizumab (TECENTRIQ ® ), avelumab (BAVENCIO ® ; MSB0010718C), envafolimab (ASC22), durvalumab (IMFINZI ®; MEDI-4736), cosibelimab (CK-301), lodalisumab (LY 3300054), galvelimab (BGB-A333), envolimab (KN035), oclacitinib (HLX-20), manelimumab (BCD-135), CX-072, CBT-502 (TQB-2450), MSB-2311, SHR-1316, sugemalimab (CS-1001; WBP3155), A167 (KL-A167, HBM 9167), STI-A1015 (IMC-001), FAZ-053, BMS-936559 (MDX1105), INCB086550, and the bispecific inhibitors GEN-1046 (PD-L1 / 4-1BB), FPT-155 (CTLA4 / PD-L1 / CD28), bintrafusp alpha (M7824; PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM-3 / PDL1), INBRX-105 (4-1BB / PDL1), MAX10181, and GNS-1480 (PD-L1 / EGFR).

[0267] In some embodiments, the small molecule PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of CA-170, GS-4224, GS-4416, INCB99280, INCB99318, and lazertinib.

[0268] Chemotherapeutic agent

[0269] In some embodiments, the chemotherapeutic agents that can be co-administered in the methods provided herein are selected from the group consisting of platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin. In some embodiments, the platinum complexes are selected from the group consisting of carboplatin, cisplatin, and oxaliplatin. In some embodiments, the taxanes are selected from the group consisting of paclitaxel, albumin-bound paclitaxel (e.g., ABRAXANE ® ), and docetaxel.

[0270] In some embodiments, the chemotherapeutic agents that can be co-administered in the methods provided herein are selected from capecitabine, cyclophosphamide, dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, epirubicin, eribulin, 5-FU, gemcitabine, irinotecan, ixabepilone, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, nab-paclitaxel, ABRAXANE ®(Paclitaxel protein-bound), pemetrexed, vinorelbine, and vincristine. In some embodiments, the chemotherapeutic agent is a kinase inhibitor. Non-limiting exemplary kinase inhibitors include erlotinib, afatinib, gefitinib, crizotinib, dabrafenib, trametinib, vemurafenib, and cobimetanib.

[0271] Breast cancer

[0272] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with breast cancer in the methods provided herein are selected from the group consisting of albumin-bound paclitaxel, anastrozole, atezolizumab, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, epirubicin, everolimus, exemestane, fluorouracil, fulvestrant, gemcitabine, ixabepilone, lapatinib, letrozole, methotrexate, mitoxantrone, paclitaxel, pegylated liposomal doxorubicin, pertuzumab, tamoxifen, toremifene, trastuzumab, vinorelbine, and any combination thereof.

[0273] Triple-negative breast cancer

[0274] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with TNBC in the methods provided herein are selected from the group consisting of cyclophosphamide, docetaxel, doxorubicin, epirubicin, fluorouracil, paclitaxel, and combinations thereof.

[0275] Colorectal cancer

[0276] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with colorectal cancer (e.g., MSS mCRC) in the methods provided herein are selected from the group consisting of capecitabine, cetuximab, fluorouracil, irinotecan, leucovorin, oxaliplatin, panitumumab, ziv-aflibercept, and any combination thereof.

[0277] Esophageal and esophagogastric junction cancer

[0278] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with esophageal or esophagogastric junction cancer in the methods provided herein are selected from the group consisting of capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, oxaliplatin, paclitaxel, and any combination thereof.

[0279] Gastric cancer

[0280] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with gastric cancer in the methods provided herein are selected from the group consisting of capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, mitomycin, oxaliplatin, paclitaxel, and any combination thereof.

[0281] Head and neck cancer

[0282] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with head and neck cancer in the methods provided herein are selected from the group consisting of afatinib, bleomycin, capecitabine, carboplatin, cetuximab, cisplatin, docetaxel, fluorouracil, gemcitabine, hydroxyurea, methotrexate, nivolumab, paclitaxel, vinorelbine, and any combination thereof.

[0283] Non-small cell lung cancer combination therapy

[0284] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with non-small cell lung cancer (NSCLC) in the methods provided herein are selected from the group consisting of afatinib, albumin-bound paclitaxel, alectinib, cabozantinib, carboplatin, cisplatin, crizotinib, dabrafenib, docetaxel, erlotinib, etoposide, gemcitabine, paclitaxel, pemetrexed, vandetanib, vemurafenib, vinblastine, vinorelbine, and any combination thereof.

[0285] Small cell lung cancer

[0286] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with small cell lung cancer (SCLC) in the methods provided herein are selected from the group consisting of bendamustine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, gemcitabine, irinotecan, paclitaxel, temozolomide, topotecan, vincristine, vinorelbine, and any combination thereof.

[0287] Ovarian cancer

[0288] In some embodiments, chemotherapeutic agents that can be co-administered to a subject with ovarian cancer in the methods provided herein are selected from the group consisting of: 5-fluorouracil, albumin-bound paclitaxel, altretamine, anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, exemestane, gemcitabine, ifosfamide, irinotecan, letrozole, leuprolide acetate, liposomal doxorubicin, megestrol acetate, melphalan, olaparib, oxaliplatin, paclitaxel, pazopanib, pemetrexed, tamoxifen, topotecan, vinorelbine, and any combination thereof.

[0289] Route of administration

[0290] In some embodiments, the anti-CCR8 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, and chemotherapeutic agent described herein can be administered in vivo by various routes, including but not limited to intravenous, intraarterial, parenteral, intratumoral, intraperitoneal, or subcutaneous. The appropriate formulation and route of administration can be selected according to the intended application.

[0291] Kit / product

[0292] The present disclosure also provides kits, medicaments, compositions, and unit dosage forms for use in any of the methods described herein.

[0293] The kit can include one or more containers that contain the anti-CCR8 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, or the chemotherapeutic agent, or unit dosage forms and / or articles. In some embodiments, unit doses are provided, wherein the unit dose contains a predetermined amount of a composition that contains the antibody and / or fusion protein provided herein, with or without one or more additional agents. In some embodiments, such unit doses are provided in a single-use prefilled syringe for injection. In some embodiments, the composition contained in the unit dose can contain saline, sucrose, etc.; buffers, such as phosphates, etc.; and / or be formulated within a stable and effective pH range. In some embodiments, the composition can be provided as a lyophilized powder that can be reconstituted upon addition of an appropriate liquid (e.g., sterile water). In some embodiments, the composition contains one or more substances that inhibit protein aggregation, including but not limited to sucrose and arginine. In some embodiments, the composition contains heparin and / or proteoglycan.

[0294] In some embodiments, the kit further comprises instructions for treating cancer according to any of the methods described herein. The kit may also include instructions for selecting an individual suitable for treatment. The instructions provided in the kit are typically written instructions on a label or package insert (e.g., a piece of paper included in the kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable. In some embodiments, the kit further comprises another therapeutic agent.

[0295] The kit is provided in a suitable package. Suitable packages include but are not limited to vials, bottles, jars, flexible packaging (e.g., sealed mylar or plastic bags), etc. The kit may optionally provide additional components, such as buffers and interpretive information. Accordingly, the present application also provides an article of manufacture comprising a vial (such as a sealed vial), bottle, jar, flexible packaging, etc.

[0296] Example

[0297] Example 1: Syngeneic mouse tumor study of anti-CCR8 antibody / chemotherapy combination therapy

[0298] In different murine syngeneic models, including 4T1 (breast cancer), Pan02 (pancreatic cancer), B16F10 (melanoma), LLC (lung cancer), the ability of the anti-CCR8 antibody to enhance tumor-specific effector T cell responses when paired with low-dose chemotherapy with or without anti-PD-1 mAb was tested in vivo. The 4T1 and Pan02 models are generally understood to represent solid tumor models with a strong immunosuppressive component. B16F10 and LLC are generally regarded as cold tumor models.

[0299] Reagent

[0300] The anti-CCR8 antibody used in the studies described herein is of the murine IgG2a isotype and is as described, for example, in Campbell, J. R. (2021). “Fc-Optimized Anti-CCR8 Antibody Depletes Regulatory T Cells in Human Tumor Models.” Cancer Res 81(11):

[0301] 2983 - 2994. The isotype control is murine IgG2a (BioXcell). The anti-PD-1 antibody is a murine IgG1 antibody with a D265A mutation.

[0302] 4T1 model

[0303] After inoculation, when the tumor reached 80 mm 3 -120 mm 3Mice were randomized (n = 8 animals / group) and administered a single dose of low-dose cisplatin at 3 mg / kg (approximately 50% of the most effective dose) and / or anti-CCR8 antibody Q3D at 1 mg / kg. The control group received an isotype control antibody. Figure 1 It was found that the anti-CCR8 antibody / cisplatin combination treatment produced stronger tumor growth inhibition than anti-CCR8 antibody or cisplatin single agent treatment, respectively.

[0304] Pan02 model

[0305] After inoculation, when the tumor reached 80 mm 3 -120mm 3 Mice were randomized (n = 8 animals / group) and administered a single dose of low-dose gemcitabine at 15 mg / kg (approximately 50% of the maximal effective dose) and / or anti-CCR8 antibody Q3D at 1 mg / kg. The control group received an isotype control antibody. Figure 2 It was found that the anti-CCR8 antibody / gemcitabine combination treatment produced stronger tumor growth inhibition than anti-CCR8 antibody or gemcitabine single agent treatment, respectively.

[0306] B16F10 model

[0307] The first study using the B16F10 model tested the relative antitumor activity of anti-CCR8 antibody and gemcitabine as single agents and in combination. 3 -120mm 3 Mice were randomized (n = 10 animals / group) and administered a single dose of low-dose gemcitabine at 15 mg / kg (approximately 50% of the maximal effective dose) and / or anti-CCR8 antibody Q3D at 1 mg / kg. The control group received an isotype control antibody. Figure 3 , Figure 4A and Figure 4B Administration of anti-CCR8 antibody and gemcitabine alone or in combination did not produce significant tumor growth inhibition ( Figure 3 ). Tumors were collected on day 5 after administration. Pharmacodynamic data showed significant Treg depletion in tumors of the anti-CCR8 antibody treatment group ( Figure 4A ), as well as significant CD8+ T cell infiltration in tumors of the chemotherapy-treated group ( Figure 4B). Briefly, tumors were dissociated and prepared for flow cytometry staining. Cells were stained as follows: L / D, CD45, CD3, CD4, CD8, CD25, FoxP3. Samples were acquired on a flow cytometer, and the data generated were used to determine the Treg frequency (L / D negative, CD45 positive, CD3 positive, CD4 positive, CD25 positive, FoxP3 positive) and the CD8 frequency (L / D negative, CD45 positive, CD3 positive, CD8 positive).

[0308] A second study using the B16F10 model tested anti-CCR8 antibody and anti-PD-1 antibody as single agents or in combination. The results are shown in Figure 5 . Treatment with the anti-CCR8 antibody / anti-PD-1 antibody combination produced significantly stronger tumor growth inhibition than single-agent treatment.

[0309] LLC model

[0310] The first study using the LLC model tested the relative anti-tumor activities of anti-CCR8 antibody and docetaxel as single agents and administered in combination. After inoculation, when tumors reached 80 mm 3 -120 mm 3 , the mice were randomly grouped (n = 8 animals / group) and administered a single dose of low-dose docetaxel at 5 mg / kg (approximately 50% of the most effective dose) and / or anti-CCR8 antibody at 1 mg / kg Q3D. The control group received an isotype control antibody. The results are shown in Figure 6 . Administration of anti-CCR8 antibody and docetaxel alone or in combination did not produce significant tumor growth inhibition ( Figure 6 ). Anti-CCR8 antibody and docetaxel were administered as single agents and in combination.

[0311] A second study using the LLC model tested the relative anti-tumor activities of anti-CCR8 antibody, anti-PD-1 antibody, and low-dose docetaxel as single agents, dual combinations, and triple combinations. After inoculation, when tumors reached 80 mm 3 -120 mm 3 , the mice were randomly grouped (n = 10 animals / group) and administered a single dose of docetaxel at 5 mg / kg, and / or anti-CCR8 antibody at 1 mg / kg Q3D, and / or anti-PD-1 antibody at 10 mg / kg Q3D. The control group received an isotype control antibody. The results of tumor growth inhibition are shown in Figure 7 . Figure 8Highlights data for individual mice in different treatment cohorts on day 15. Treatment with each anti-CCR8 antibody, anti-PD-1 antibody, and docetaxel monotherapy resulted in partial tumor growth inhibition. Treatment with anti-CCR8 antibody / docetaxel, anti-PD-1 antibody / docetaxel, and anti-CCR8 antibody / anti-PD-1 antibody dual combinations resulted in improved tumor growth inhibition. Treatment with anti-CCR8 antibody / anti-PD-1 antibody / docetaxel triple combination resulted in further improved tumor growth inhibition.

[0312] Conclusion

[0313] This example demonstrates that targeting Treg depletion in combination with chemotherapy can result in significantly reduced tumor growth in breast and pancreatic tumor models with strong immunosuppressive components ( Figure 1 and Figure 2 ). No efficacy of the chemotherapy and anti-CCR8 mAb combination was observed in models with low immune infiltration ( Figure 3 ), although intratumoral Treg depletion was confirmed ( Figure 4A ), and Teff infiltration was observed ( Figure 4B ). In the presence of PD-1 blockade, selective Treg depletion was found to generate a strong response in cold tumor models ( Figure 5 ). Addition of anti-PD-1 antibody to low-dose chemotherapy and Treg depletion in the LLC model (which is non-responsive to single-agent or combination therapies, i.e., anti-CCR8 antibody and chemotherapy, Figure 6 ) resulted in 72% tumor growth inhibition in the triple combination group ( Figure 7 and Figure 8 ), indicating that PD-1 expression can limit T cell responses even in the absence of Tregs.

[0314] In summary, this example demonstrates that Treg depletion can enhance the response to chemotherapy treatment, including low-dose chemotherapy treatment. PD-1 blockade can potentiate the effects of selective Treg depletion and chemotherapy to generate strong anti-tumor immune activity.

[0315] * * * * *

[0316] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and those skilled in the art will make various modifications or changes based on these examples and embodiments and include such modifications or changes within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0317] Table of certain sequences

[0318]

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

Claims

1. A method of treating cancer in a subject, the method comprising co-administering to the subject an effective amount of: i) an anti-CCR8 antibody; ii) a chemotherapeutic agent, and iii) a PD-1 inhibitor or a PD-L1 inhibitor; wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody.

2. The method according to claim 1, wherein the chemotherapeutic agent is administered at a lower dose than in a standard of care chemotherapy regimen that does not include an anti-CCR8 antibody.

3. A method of treating cancer in a subject, the method comprising co-administering to the subject an effective amount of: i) an anti-CCR8 antibody; ii) a chemotherapeutic agent, and iii) optionally a PD-1 inhibitor or a PD-L1 inhibitor; wherein the anti-CCR8 antibody has antibody-dependent cellular cytotoxicity (ADCC) activity and / or complement-dependent cytotoxicity (CDC) activity; wherein the anti-CCR8 antibody is optionally a CCR8-neutralizing antibody, and wherein the chemotherapeutic agent is administered at a lower dose than in a standard of care chemotherapy regimen that does not include an anti-CCR8 antibody.

4. The method according to any one of claims 1 to 3, wherein the chemotherapeutic agent is a single chemotherapeutic agent.

5. The method according to any one of claims 1 to 3, wherein the chemotherapeutic agent is a plurality of chemotherapeutic agents.

6. The method according to any one of claims 1 to 5, wherein the chemotherapeutic agent is selected from the group consisting of: platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin.

7. The method according to claim 6, wherein the chemotherapeutic agent comprises a platinum complex.

8. The method according to claim 6 or 7, wherein the platinum complex is selected from the group consisting of: carboplatin, cisplatin, and oxaliplatin.

9. The method according to claim 6, wherein the chemotherapeutic agent comprises a taxane.

10. The method according to claim 9, wherein the taxane is docetaxel.

11. The method according to claim 6, wherein the chemotherapeutic agent comprises gemcitabine.

12. The method according to any one of claims 1 to 11, wherein the dose of the chemotherapeutic agent is 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 50% or less, 40% or less, 30% or less, or 20% or less of the dose of the chemotherapeutic agent administered in a standard of care regimen that does not include an anti-CCR8 antibody.

13. The method according to any one of claims 1 to 12, wherein the cancer comprises solid tumors.

14. The method according to claim 13, wherein the cancer comprises tumor-infiltrating Treg cells that express CCR8.

15. The method according to claim 14, wherein CCR8 is expressed on the surface of the Treg cells at less than 10,000 copies / cell (as determined by fluorescence-activated cell sorting (FACS) and / or flow cytometry).

16. The method according to any one of claims 1 to 15, wherein the cancer is selected from the group consisting of: breast cancer, colorectal cancer, head and neck cancer, lung cancer, ovarian cancer, gastric cancer, gastric adenocarcinoma, and thymoma.

17. The method according to any one of claims 1 to 15, wherein the cancer is selected from the group consisting of: endometrial adenocarcinoma, colorectal cancer, ovarian cancer, vaginal squamous cell carcinoma, endometrial adenocarcinoma, colorectal cancer, melanoma (e.g., cutaneous melanoma), pancreatic cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), uterine leiomyosarcoma, cholangiocarcinoma, adenoid cystic carcinoma, cervical cancer, renal cell carcinoma (RCC), anal cancer, esophagogastric junction (EGJ) adenocarcinoma, and gastric adenocarcinoma.

18. The method according to claim 16 or 17, wherein the cancer is ovarian cancer and the chemotherapeutic agent is selected from the group consisting of: 5-fluorouracil, albumin-bound paclitaxel, altretamine, anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, exemestane, gemcitabine, ifosfamide, irinotecan, letrozole, leuprorelin acetate, liposomal doxorubicin, megestrol acetate, melphalan, olaparib, oxaliplatin, paclitaxel, pazopanib, pemetrexed, tamoxifen, topotecan, vinorelbine, and any combination thereof.

19. The method according to any one of claims 1 to 15, wherein the cancer is selected from the group consisting of: head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer (NSCLC), gastric adenocarcinoma, EGJ adenocarcinoma, and colorectal cancer (CRC) (e.g., MSS mCRC).

20. The method according to claim 19, wherein the cancer is HNSCC and the co-administered chemotherapeutic agent is selected from the group consisting of: afatinib, bleomycin, capecitabine, carboplatin, cetuximab, cisplatin, docetaxel, fluorouracil, gemcitabine, hydroxyurea, methotrexate, nivolumab, paclitaxel, vinorelbine, and any combination thereof.

21. The method according to claim 19, wherein the cancer is gastric adenocarcinoma and the co-administered chemotherapeutic agent is selected from the group consisting of: capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, mitomycin, oxaliplatin, paclitaxel, and any combination thereof.

22. The method according to claim 19, wherein the cancer is adenocarcinoma of the esophagogastric junction (EGJ) and the co-administered chemotherapeutic agent is selected from the group consisting of capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, oxaliplatin, paclitaxel, and any combination thereof.

23. The method according to claim 19, wherein the cancer is colorectal cancer and the co-administered chemotherapeutic agent is selected from the group consisting of capecitabine, cetuximab, fluorouracil, irinotecan, leucovorin, oxaliplatin, panitumumab, ziv-aflibercept, and any combination thereof.

24. The method according to any one of claims 1 to 15, wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, and lung cancer.

25. The method according to claim 24, wherein the cancer is breast cancer and the co-administered chemotherapeutic agent is selected from the group consisting of albumin-bound paclitaxel, anastrozole, atezolizumab, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, epirubicin, everolimus, exemestane, fluorouracil, fulvestrant, gemcitabine, ixabepilone, lapatinib, letrozole, methotrexate, mitoxantrone, paclitaxel, pegylated liposomal doxorubicin, pertuzumab, tamoxifen, toremifene, trastuzumab, vinorelbine, and any combination thereof.

26. The method according to claim 24, wherein the breast cancer is selected from triple-negative breast cancer (TNBC), HR+ / HER2-breast cancer, or HR+ / HER-low breast cancer.

27. The method according to claim 26, wherein the breast cancer is TNBC and the chemotherapeutic agent is selected from the group consisting of cyclophosphamide, docetaxel, doxorubicin, epirubicin, fluorouracil, paclitaxel, and any combination thereof.

28. The method according to claim 24, wherein the pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC).

29. The method according to claim 16 or 24, wherein the cancer is lung cancer.

30. The method according to claim 29, wherein the lung cancer is non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC).

31. The method according to claim 30, wherein the lung cancer is NSCLC.

32. The method according to claim 30, wherein the lung cancer is NSCLC and the co-administered chemotherapeutic agent is selected from the group consisting of afatinib, albumin-bound paclitaxel, alectinib, cabozantinib, carboplatin, cisplatin, crizotinib, dabrafenib, docetaxel, erlotinib, etoposide, gemcitabine, paclitaxel, pemetrexed, vandetanib, vemurafenib, vinblastine, vinorelbine, and any combination thereof.

33. The method according to claim 30, wherein the lung cancer is SCLC and the co-administered chemotherapeutic agent is selected from the group consisting of: 5-fluorouracil, albumin-bound paclitaxel, altretamine, anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, exemestane, gemcitabine, ifosfamide, irinotecan, letrozole, leuprorelin acetate, liposomal doxorubicin, megestrol acetate, melphalan, olaparib, oxaliplatin, paclitaxel, pazopanib, pemetrexed, tamoxifen, topotecan, vinorelbine, and any combination thereof.

34. The method according to any one of claims 1 to 33, wherein the cancer is metastatic.

35. The method according to any one of claims 1 to 12, wherein the cancer is a blood cancer expressing CCR8.

36. The method according to claim 35, wherein the blood cancer is selected from the group consisting of: T-cell adult acute lymphoblastic leukemia, T-cell pediatric acute lymphoblastic leukemia, lymphoblastic lymphoma, acute lymphoblastic leukemia, cutaneous T-cell lymphoma (CTCL), T-cell acute lymphoblastic leukemia, adult T-cell leukemia / lymphoma, T-cell lymphoblastic leukemia / lymphoma, and anaplastic large cell lymphoma.

37. The method according to claim 36, wherein the blood cancer is CTCL.

38. The method according to any one of claims 1 to 37, wherein the subject is a human.

39. The method according to any one of claims 1 to 38, wherein the subject has not received treatment.

40. The method according to any one of claims 1 to 38, wherein the subject has received one or more courses of anti-cancer treatment, and optionally wherein the cancer has progressed during one or more courses of anti-cancer treatment.

41. The method according to any one of claim 40, wherein the anti-cancer treatment is selected from the group consisting of: surgery, radiotherapy, hormone therapy, targeted anti-cancer agents, chemotherapeutic agents, immunotherapy, and antibody-drug conjugates (ADCs).

42. The method according to claim 41, wherein the chemotherapeutic agent is selected from the group consisting of: platinum complexes, taxanes, pemetrexed, gemcitabine, fluorouracil, irinotecan, etoposide, and doxorubicin.

43. The method according to claim 42, wherein the platinum complex is selected from the group consisting of: carboplatin, cisplatin, and oxaliplatin.

44. The method according to claim 41, wherein the immunotherapy comprises an anti-PD-1 antibody or an anti-PD-L1 antibody.

45. The method of claim 44, wherein the anti-PD-1 antibody or the anti-PD-L1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiprilimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cocilimab, sazanlimab, tislelizumab, rivolimab, batilizumab, toripalimab, sitrilimab, genoluzumab, palolizumab, lodalizumab, carrelizumab, budigalimab, avelumab, dotalimumab, envolimab, sintilimab and sepalizumab.

46. ​​The method of claim 44 or 45, wherein the immunotherapy further comprises an anti-TIGIT antibody.

47. The method of claim 46, wherein the anti-TIGIT antibody is selected from the group consisting of tisleliumab, vembryomab, dunalimab, AB308, AK127, BMS-986207, or etilizumab.

48. The method of any one of claims 1 to 47, wherein the anti-CCR8 antibody comprises: i) HCDR1 comprising the amino acid sequence of SEQ ID NO: 12, HCDR2 comprising the amino acid sequence of SEQ ID NO: 13, HCDR3 comprising the amino acid sequence of SEQ ID NO: 14, LCDR1 comprising the amino acid sequence of SEQ ID NO: 15, LCDR2 comprising the amino acid sequence of SEQ ID NO: 16, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 17; ii) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 25, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 26, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 27, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 29; iii) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 36, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 37, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 39, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 40, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 41; iv) An HCDR1 comprising the amino acid sequence of SEQ ID NO: 48, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 49, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 50, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 51, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 52, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 53; v) An HCDR1 comprising the amino acid sequence of SEQ ID NO: 60, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 61, 72 or 78, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 62, 73 or 79, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 63, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 64, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 65; or vi) An HCDR1 comprising the amino acid sequence of SEQ ID NO: 84 or 100, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 85, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 86, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 87, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 88, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

89.

49. The method according to any one of claims 1 to 47, wherein the anti-CCR8 antibody comprises: i) A heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 68 or 74, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 69 or 75; or ii) A heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 92 or 96, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 93 or 97.

50. The method according to any one of claims 1 to 47, wherein the anti-CCR8 antibody comprises: i) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 68 or 74, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 69 or 75; or ii) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 92 or 96, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 93 or 97.

51. The method according to any one of claims 48 to 50, wherein the anti-CCR8 antibody is a monoclonal antibody.

52. The method according to any one of claims 48 to 51, wherein the anti-CCR8 antibody is a humanized antibody.

53. The method according to any one of claims 48 to 51, wherein the anti-CCR8 antibody is a full-length antibody.

54. The method according to any one of claims 48 to 51, wherein the anti-CCR8 antibody is an IgG1 or IgG3 antibody.

55. The method according to any one of claims 1 to 54, wherein the anti-CCR8 antibody comprises: i) a heavy chain (HC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 70 or 76, and a light chain (LC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 71 or 77; or ii) a heavy chain (HC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 94 or 98, and a light chain (LC) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 95 or 99.

56. The method according to claim 55, wherein the anti-CCR8 antibody comprises: i) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 70 or 76, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 71 or 77; or ii) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 94 or 98, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 95 or 99.

57. The method according to any one of claims 1 to 56, wherein the anti-CCR8 antibody is an afucosylated antibody.

58. The antibody according to any one of claims 1 to 56, wherein the anti-CCR8 antibody comprises a heavy chain constant region mutation at one or more positions selected from L234, L235, G236, S239, F243, H268, D270, R292, S298, Y300, V305, K326, A330, I332, E333, K334, and P396.

59. The antibody according to any one of claims 1 to 56, wherein the anti-CCR8 antibody comprises a heavy chain constant region mutation selected from S239D, S239M, F243L, H268D, D270E, R292P, S298A, Y300L, V305I, K326D, A330L, A330M, I332E, E333A, K334A, K334E, and P396L.

60. The antibody according to any one of claims 1 to 56, wherein the anti-CCR8 antibody comprises a heavy chain constant region mutation selected from F243L / R292P / Y300L / V305I / P396L, S239D / I332E, S239D / I332E / A330L, S298A / E333A / K334A, L234Y / L235Q / G236W / S239M / H268D / D270E / S298A, and D270E / K326D / A330M / K334E.

61. The method according to any one of claims 1 to 56, wherein the anti-CCR8 antibody inhibits the binding of CCL1 to CCR8.

62. The method according to any one of claims 1 to 61, wherein the anti-CCR8 antibody is selected from the group consisting of BMS-986340 (Bristol Myers Squibb), LM-108 (LaNova Medicines), S-531011 (Shionogi), FPA157 (Five Prime, Amgen), IPG-7236 (Immunophage Biomedical), ICP-B05 (InnoCare Pharma Tech), SRF-114 (Surface Oncology), HBM1022 (Harbour BioMed), HFB1011 (HiFiBio), BAY-3375968 (Bayer), IO-1 (Oncurious), ZL-1218 (Zai Lab), GB2101 (Genor), and PSB-114 (Sound Biologics).

63. The method according to any one of claims 1 to 62, wherein the co-administered PD-1 inhibitor or PD-L1 inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody.

64. The method according to claim 63, wherein the anti-PD-1 antibody or anti-PD-L1 antibody is selected from the group consisting of: pembrolizumab, nivolumab, cemiplimab, pidilizumab, spartalizumab, atezolizumab, avelumab, durvalumab, cosibelimab, sasanlimab, tislelizumab, rivulizumab, batrilizumab, toripalimab, cetrelimab, genolimab, palovelimab, lodavalimab, camrelizumab, budigalimab, avelumab, dostarlimab, envafolimab, sintilimab, and sepantronium.

65. The method according to any one of claims 1 to 62, wherein the PD-1 inhibitor or the PD-L1 inhibitor is a small molecule inhibitor.

66. The method according to claim 65, wherein the small molecule PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of: CA-170, GS-4224, GS-4416, INCB99280, INCB99318, and lazertinib.

67. The method according to any one of claims 1 to 66, further comprising administering to the subject one or more additional therapeutic agents.

68. An anti-CCR8 antibody for use in combination with a chemotherapeutic agent and a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) in a method of treating cancer, wherein the method comprises administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the PD-1 inhibitor or the PD-L1 inhibitor (e.g., an anti-PD1 antibody or an anti-PD-L1 antibody), wherein the anti-CCR8 antibody has antibody-dependent cell cytotoxicity (ADCC) activity and / or complement-dependent cell cytotoxicity (CDC) activity, and wherein the anti-CCR8 antibody is optionally a CCR8 neutralizing antibody.

69. An anti-CCR8 antibody for use in combination with a chemotherapeutic agent and optionally a PD-1 inhibitor or a PD-L1 inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) in a method of treating cancer, wherein the method comprises administering to a subject the anti-CCR8 antibody, the chemotherapeutic agent, and the PD-1 inhibitor or the PD-L1 inhibitor (e.g., an anti-PD1 antibody or an anti-PD-L1 antibody), wherein the anti-CCR8 antibody has antibody-dependent cell cytotoxicity (ADCC) activity and / or complement-dependent cell cytotoxicity (CDC) activity, wherein the anti-CCR8 antibody is optionally a CCR8 neutralizing antibody, and wherein the chemotherapeutic agent is administered at a lower dose than in a standard of care chemotherapy regimen that does not include an anti-CCR8 antibody-based agent.

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