Methods of treating cancer using ilt2 antibodies and combination therapies

Abstract

Methods for treating cancer using antibodies that specifically bind to human ILT2, and combinations of these antibodies with human CTLA-4 and / or human PD-1 inhibitors, are provided. These methods are particularly advantageous in that the anti-human ILT2 antibodies described herein effectively reverse ILT2-mediated immunosuppression and promote the activation of NK cells, NKT cells, T cells, and myeloid cells.

Description

[0001] Related applications This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 582,093, filed September 12, 2023, the entire disclosure of which is incorporated herein by reference. Technical Field

[0002] This disclosure relates to methods of treating cancer using antibodies that specifically bind to human ILT2, and combination therapies with CTLA-4 and / or PD-1 inhibitors. Background Technology

[0003] ILT2 is an inhibitory receptor belonging to the class I transmembrane glycoproteins. It possesses four extracellular immunoglobulin-like domains (D1-D4), a transmembrane region, and an intracellular tail region with four immunoreceptor tyrosine-based inhibitory motifs (ITIMs). ILT2 is expressed on a variety of immune cells, such as T cells, B cells, natural killer (NK) cells, myeloid-derived suppressor cells (MDSCs), dendritic cells (DCs), and subsets of monocytes / macrophages. Furthermore, ILT2 binds to both canonical (HLA-A and -B) and atypical (HLA-G, HLA-E, and HLA-F) major histocompatibility complex (MHC) class I molecules. High ILT2 expression is associated with poor prognosis in several human malignancies, and ILT2 activation has been reported to impair the cytotoxic activity of NK cells and effector T cells, attenuate B cell function, inhibit antigen presentation by dendritic cells, and promote immunosuppressive activity of myeloid cells.

[0004] CTLA-4 is an inhibitory receptor upregulated on T cells (Alegre et al., 2001, Nat RevImmunol 1:220-8). CTLA-4 suppresses immune responses in several ways: it competes with the T cell costimulatory receptor CD28 for its ligands CD80 and CD86, thereby blocking costimulation; it inhibits T cell activation through negative signaling; and it can also take up CD80 and CD86 from opposing cells via trans-endocytosis, leading to impaired T cell costimulation via CD28 (Krummel and Allison, 1995, J Exp Med 182:459-465; Walunas et al., 1994, Immunity 1:405-413; Qureshi et al., 2011, Science 332:600-603).

[0005] PD-1 is another inhibitory receptor expressed on activated B cells, T cells, and myeloid cells (Agata et al., (1996) Int Immunol 8:765-72; Okazaki et al., (2002) Curr. Opin. Immunol. 14: 391779-82; Bennett et al., (2003) J Immunol 170:711-8). Two ligands of PD-1, PD-L1 and PD-L2, have been identified, and these ligands have been shown to downregulate T cell activation upon binding to PD-1 (Freeman et al., (2000) J Exp Med 192: 1027-34; Latchman et al., (2001) Nat Immunol 2:261-8; Carter et al., (2002) Eur J Immunol 32:634-43). The interaction between PD-1 and PD-L1 / PD-L2 leads to a reduction in tumor-infiltrating lymphocytes, weakened T-cell receptor-mediated proliferation, and immune escape from cancer cells (Dong et al., (2003) J.Mol. Med. 81:281-7; Blank et al., (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al., (2004) Clin. Cancer Res. 10:5094-100). This immunosuppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1 (Iwai et al., (2002) Proc. Natl Acad. Sci. USA 99:12293-7; Brown et al., (2003) J. Immunol. 170: 1257-66).

[0006] Given the important roles of ILT2, CTLA-4, and PD-1 in regulating immune responses, therapies aimed at antagonizing signal transduction from these receptors show great promise in enhancing anti-cancer immunity and effectively treating cancer. Summary of the Invention

[0007] This disclosure provides methods for treating cancer using antibodies that specifically bind to human ILT2, and combinations of these antibodies with human CTLA-4 and / or human PD-1 inhibitors. These methods are particularly advantageous in that the anti-human ILT2 antibodies described herein effectively reverse ILT2-mediated immunosuppression and promote the activation of NK cells, NKT cells, T cells, and myeloid cells. Therapeutic modulation of the co-inhibitory receptors of ILT2, PD-1, and CTLA-4 effectively enhances anti-cancer immunity and overcomes tumor immunosuppression to treat cancer.

[0008] Therefore, in one aspect, this disclosure provides a method for treating cancer in a subject in need, the method comprising administering to the subject a first antibody that specifically binds to human ILT2, the first antibody being in a dose of about 1 mg to about 2000 mg, wherein the first antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8.

[0009] In another aspect, this disclosure provides a method of treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human PD-1 inhibitor. In some embodiments, the human PD-1 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

[0010] In another aspect, this disclosure provides a method of treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human CTLA-4 inhibitor. In some embodiments, the human CTLA-4 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

[0011] In another aspect, this disclosure provides a method of treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human PD-L1 inhibitor. In some embodiments, the human PD-L1 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

[0012] In another aspect, this disclosure provides a method of treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human PD-L2 inhibitor. In some embodiments, the human PD-L2 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

[0013] In another aspect, this disclosure provides a method for treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; a human PD-1 inhibitor; and a human CTLA-4 inhibitor.

[0014] In some embodiments, the first antibody is administered at a dose of about 1 mg to about 1000 mg. In some embodiments, the first antibody is administered at a dose of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, or about 1000 mg.

[0015] In some embodiments, the first antibody is administered intravenously. In some embodiments, the first antibody is administered via intravenous infusion over approximately 30 minutes.

[0016] In some embodiments, the primary antibody is administered weekly. In some embodiments, the primary antibody is administered every two weeks. In some embodiments, the primary antibody is administered every three weeks. In some embodiments, the primary antibody is administered every four weeks. In some embodiments, the primary antibody is administered every five weeks. In some embodiments, the primary antibody is administered every six weeks.

[0017] In some embodiments, the primary antibody is administered intravenously at a dose of 1 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 3 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 10 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 30 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 1000 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 300 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 1000 mg every 3 weeks.

[0018] In some implementations, the cancer is a solid tumor. In some implementations, the solid tumor is metastatic or locally advanced. In some implementations, the cancer is selected from the group consisting of: bladder cancer, brain cancer, breast cancer, cervical cancer, colon and / or rectal cancer, endometrial cancer, head and neck cancer, kidney cancer, liver cancer, melanoma, mesothelioma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer, small cell lung cancer, gastric cancer, and thyroid cancer.

[0019] In some implementation schemes, prior to administration of the primary antibody: the subject is at least 18 years old; the subject is histologically diagnosed with a currently metastatic or locally advanced solid tumor for which no standard therapy is available or has failed; the subject has measurable disease on baseline imaging, as assessed according to RECIST version 1.1; and the subject's life expectancy is [not specified]. > 3 months; the subject's Eastern Oncology Cooperative Group performance status score was 0-1; the subject had adequate organ and bone marrow reserve, as defined by one or more of the following: absolute neutrophil count > 1.5×10 9 / L; platelet count > 100×10 9 / L; Hemoglobin > 8.0 g / dL, and no blood transfusion within 2 weeks prior to hemoglobin measurement; serum albumin > 3.0 g / dL; Total bilirubin < 1.5 × Upper Limit of Normal (ULN); Aspartate Aminotransferase <2.5×ULN; alanine aminotransferase < 2.5×ULN; alkaline phosphatase < 2.5×ULN, or for subjects with liver metastases < 5.0 × ULN; Creatinine clearance > 40 mL / min, as assessed by the Cockcroft-Gault method; and / or international normalized ratio or prothrombin time. < 1.5×ULN and activation time of partial thromboplastin < 1.5×ULN, unless the patient is receiving anticoagulation therapy; the subject is not pregnant and / or breastfeeding; the subject has not received cytotoxic therapy or other monoclonal antibody therapy within 3 weeks prior to enrollment; the subject has not received central nervous system (CNS) radiation therapy within 2 weeks prior to enrollment or non-CNS radiation therapy within 1 week prior to enrollment; and / or the subject has not received small molecule targeted therapy or tyrosine kinase inhibitor therapy within 2 weeks or 5 half-lives prior to enrollment; the subject has no persistent toxicity from previous cancer therapies (Common Terminology Criteria for Adverse Events [CTCAE]). > Grade 1); Subjects do not have grade 2 sensory neuropathy or alopecia; Subjects do not have a history of interstitial lung disease or active interstitial lung disease; Subjects do not have a history of allergic reactions or uncontrolled asthma; Subjects do not have a condition requiring systemic treatment with corticosteroids (daily prednisone equivalent >10 mg) within 2 weeks prior to enrollment or other immunosuppressive drugs within 30 days prior to enrollment; Subjects do not have active CNS metastases unless the CNS metastases have been treated and the subject is radiographically and clinically stable; Subjects do not have an active autoimmune disease or a history of autoimmune disease requiring systemic treatment within 2 years prior to enrollment, unless the treatment includes hormone replacement therapy or topical treatment; Subjects have not received allogeneic tissue or solid organ transplants, except for corneal transplants; Subjects do not have an active infection requiring treatment within 2 weeks prior to enrollment; Subjects are not HIV positive unless the subject's CD4 count is >200 cells / mm². 3 Or undetectable viral load; the subject does not have active hepatitis B (HBV) or hepatitis C (HPC) unless their hepatitis B surface antigen and HBV DNA or HCV RNA are negative; the subject does not have clinically significant cardiovascular disease, as defined by one or more of the following: cerebrovascular accident or stroke or myocardial infarction within 6 months prior to enrollment; unstable angina; congestive heart failure (New York Heart Association classification). >Grade II); and / or uncontrolled arrhythmias requiring medication; the subject's QTc interval is not greater than 480 milliseconds, unless the QTc prolongation is due to right bundle branch block; and / or the subject does not have uncontrolled hypertension (>140 / 90 mmHg), or does not have hypertension requiring more than 3 antihypertensive medications.

[0020] In some embodiments, the first antibody comprises the amino acid sequences CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 as shown in SEQ ID NO: 1, 2, 3, 4, 5, and 6, respectively. In some embodiments, the first antibody comprises the VH amino acid sequence of SEQ ID NO: 7, and / or the VL amino acid sequence of SEQ ID NO: 8. In some embodiments, the first antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 7 and 8, respectively. In some embodiments, the first antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM. In some embodiments, the heavy chain constant region is the human IgG4 heavy chain constant region, which contains proline at position 228 according to the EU numbering system. In some embodiments, the first antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the first antibody comprises a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 9 and 10, respectively.

[0021] In some embodiments, the human PD-1 inhibitor is a second antibody that specifically binds to human PD-1, human PD-L1, or human PD-L2. In some embodiments, the second antibody comprises: VH, which comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 17; and VL, which comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the second antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences shown in SEQ ID NO: 11, 12, 13, 14, 15, and 16, respectively. In some embodiments, the second antibody comprises the VH amino acid sequence of SEQ ID NO: 17 and / or the VL amino acid sequence of SEQ ID NO: 18. In some embodiments, the second antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 17 and 18, respectively. In some embodiments, the second antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM. In some embodiments, the heavy chain constant region is the human IgG4 heavy chain constant region, which contains proline at position 228 according to the EU numbering system. In some embodiments, the second antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 19, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the second antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 19 and 20, respectively.

[0022] In some implementations, the second antibody is selected from the group consisting of: balstilimab, adebrelimab, atezolizumab, avelumab, camrelizumab, cemiplimab, cosibelimab, dostarlimab, durvalumab, enlonstobart, envafolimab, nivolumab, and pembrolizumab. The following are listed: olizumab, penpulimab, pidilizumab, prolgolimab, puctenlimab, retifanlimab, serplulimab, sintilimab, socazolimab, sugemalimab, tagitanlimab, tislelizumab, toripalimab, and zimberelimab.

[0023] In some embodiments, the second antibody is administered at a dose of approximately 450 mg. In some embodiments, the second antibody is administered at a dose of approximately 3 mg / kg.

[0024] In some embodiments, the second antibody is administered via intravenous infusion over approximately 30 minutes. In some embodiments, the second antibody is administered once weekly. In some embodiments, the second antibody is administered once every two weeks. In some embodiments, the second antibody is administered once every three weeks. In some embodiments, the second antibody is administered to the subject after the administration of the first antibody.

[0025] In some embodiments, the human CTLA-4 inhibitor is a third antibody that specifically binds to human CTLA-4. In some embodiments, the third antibody comprises: VH, which comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 27; and VL, which comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 28. In some embodiments, the third antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences shown in SEQ ID NO: 21, 22, 23, 24, 25, and 26, respectively. In some embodiments, the third antibody comprises the VH amino acid sequence of SEQ ID NO: 27 and / or the VL amino acid sequence of SEQ ID NO: 28. In some embodiments, the third antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 27 and 28, respectively. In some embodiments, the third antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM. In some embodiments, the third antibody comprises a human IgG1 heavy chain constant region that is a variant of the wild-type human IgG1 heavy chain constant region, wherein the variant human IgG1 heavy chain constant region has a higher binding affinity for FcγRIIIA than the wild-type human IgG1 heavy chain constant region. In some embodiments, the third antibody is unfucosylated. In some embodiments, the heavy chain constant region is the human IgG1 heavy chain constant region comprising aspartic acid at position 239, leucine at position 330, and glutamic acid at position 332 according to the EU numbering system. In some embodiments, the third antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 29, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 30. In some embodiments, the third antibody comprises a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 29 and 30, respectively. In some embodiments, the third antibody is selected from the group consisting of botensilimab, ipilimumab, and tremelimumab.

[0026] In some embodiments, the third antibody is administered at a dose of about 150 mg. In some embodiments, the third antibody is administered at a dose of about 75 mg. In some embodiments, the third antibody is administered at a dose of about 50 mg. In some embodiments, the third antibody is administered at a dose of about 2 mg / kg. In some embodiments, the third antibody is administered at a dose of about 4 mg / kg.

[0027] In some embodiments, the third antibody is administered via intravenous infusion over approximately 30 minutes. In some embodiments, the third antibody is administered every 2 weeks. In some embodiments, the third antibody is administered every 3 weeks. In some embodiments, the third antibody is administered every 4 weeks. In some embodiments, the third antibody is administered every 6 weeks.

[0028] In some embodiments, the third antibody is administered to the subject after the administration of the first antibody. In some embodiments, the third antibody is administered to the subject after the administration of the first and second antibodies.

[0029] In another aspect, this disclosure provides a method of treating cancer in a subject in need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 9 and 10, respectively; baritelimab administered by intravenous infusion at a dose of 450 mg every 3 weeks; and botelimab administered by intravenous infusion at a dose of 150 mg every 6 weeks.

[0030] In some embodiments, the first antibody is administered at a dose of about 1 mg to about 1000 mg. In some embodiments, the first antibody is administered at a dose of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, or about 1000 mg.

[0031] In some embodiments, the first antibody is administered intravenously. In some embodiments, the first antibody is administered via intravenous infusion over approximately 30 minutes.

[0032] In some embodiments, the primary antibody is administered weekly. In some embodiments, the primary antibody is administered every two weeks. In some embodiments, the primary antibody is administered every three weeks. In some embodiments, the primary antibody is administered every four weeks. In some embodiments, the primary antibody is administered every five weeks. In some embodiments, the primary antibody is administered every six weeks.

[0033] In some embodiments, the primary antibody is administered intravenously at a dose of 1 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 3 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 10 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 30 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 1000 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 300 mg every 3 weeks. In some embodiments, the primary antibody is administered intravenously at a dose of 1000 mg every 3 weeks.

[0034] In some implementations, the cancer is a solid tumor. In some implementations, the solid tumor is metastatic or locally advanced. In some implementations, the cancer is selected from the group consisting of: bladder cancer, brain cancer, breast cancer, cervical cancer, colon and / or rectal cancer, endometrial cancer, head and neck cancer, kidney cancer, liver cancer, melanoma, mesothelioma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer, small cell lung cancer, gastric cancer, and thyroid cancer.

[0035] In another aspect, this disclosure provides an antibody that specifically binds to human ILT2 for use in the treatment of cancer, wherein the treatment is performed according to the method of any one of the preceding claims.

[0036] In another aspect, this disclosure provides an antibody that specifically binds to human ILT2 for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the method of any one of the preceding claims.

[0037] In another aspect, this disclosure provides the use of an antibody that specifically binds to human ILT-2 for the treatment of cancer, wherein the treatment is performed according to the method of any one of the preceding claims. Attached Figure Description

[0038] Figures 1A to 1D Exemplary data depicting the additive effects of the combination of anti-ILT-2 AGEN1571 and anti-PD-1 AGEN2034 (batéliximab) on immune cell activation are presented. In short, PBMCs were stimulated with IL-2 and IL-15 in the presence of (i) AGEN1571, (ii) AGEN2034, (iii) the combination of AGEN1571 and AGEN2034, or (iv) an isotype control antibody, followed by stimulation with HLA-G-expressing JEG-3. HLA-A2 Tumor cells were co-cultured for 72 hours. NK cells were detected by flow cytometry. Figure 1A ), NKT cells ( Figure 1B ) and CD8+ T cells ( Figure 1C CD25 + The frequency of TNFα was plotted using GraphPad Prism. An FMO control was used to set the gating. Data from 10 donors tested in 3 independent experiments are shown. TNFα was analyzed using a Luminex immunoassay. Figure 1D ), IFNγ ( Figure 1E ) and MIP-1β ( Figure 1F Statistical analysis was performed using one-way ANOVA. A p-value <0.05 was considered statistically significant and was used to represent differences between groups (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0039] Figures 2A to 2B An exemplary dose-response curve illustrating the additive effect of the combination of anti-ILT2 AGEN1571 and anti-CTLA-4 AGEN1181 (boretolimab) was plotted. In short, engineered Jurkat ILT2 CD16 NFAT luciferase cells were co-cultured with Raji-CTLA-4 cells in the presence of both AGEN1181 and AGEN1571. Figure 2A and Figure 2B The same data is plotted in the figure, with the AAGEN1181 dose titration shown on the x-axis. Figure 2A ) or AGEN 1571 titration display on the x-axis ( Figure 2B The IgG4 and IgG1.DLE isotype controls are also shown. The luciferase activity of the NFAT reporter gene induced by the activity of Fcγ receptor IIIA (CD16) was measured and expressed as relative luminescent units (RLU). Detailed Implementation

[0040] This disclosure provides methods for treating cancer using antibodies that specifically bind to human ILT2, and combinations of these antibodies with human CTLA-4 and / or human PD-1 inhibitors. These methods are particularly advantageous because the anti-human ILT2 antibodies described herein are potent blockers of ILT2-HLA-G interaction and have been shown to enhance immune cell function, including increased macrophage activation. Therapeutic modulation of the co-inhibitory receptors of ILT2, PD-1, and CTLA-4 effectively enhances anti-cancer immunity and overcomes tumor immunosuppression for cancer treatment.

[0041] As used herein, the terms “about” and “approximately” when used to modify numerical values ​​or ranges indicate that deviations of 5% to 10% above and below the value or range are still within the intended meaning of the value or range.

[0042] As used herein, “ILT2” refers to immunoglobulin-like transcript 2, also known as leukocyte immunoglobulin-like receptor subfamily B member 1 (LILRB1) or leukocyte immunoglobulin-like receptor 1 (LIR-1). As used herein, the term “human ILT2” refers to the human ILT2 protein encoded by the wild-type ILT2 gene (e.g., RefSeq accession number NC_000019.10). An exemplary immature amino acid sequence of human ILT2 is provided in RefSeq accession number NP_006660.4.

[0043] As used herein, the term “CTLA-4” refers to cytotoxic T-lymphocyte-associated protein 4. As used herein, the term “human CTLA-4” refers to the human CTLA-4 protein encoded by the wild-type human CTLA-4 gene (e.g., RefSeq accession number NM_005214.5 or NM_001037631.2). An exemplary immature amino acid sequence of human CTLA-4 is shown in RefSeq accession number NP_005205.2.

[0044] As used herein, the term "PD-1" refers to programmed cell death protein 1. As used herein, the term "human PD-1" refers to the human PD-1 protein encoded by the wild-type human PD-1 gene (e.g., RefSeq accession number NM_005018.3). An exemplary immature amino acid sequence of human PD-1 is provided in RefSeq accession number NP_005009.2.

[0045] As used herein, the term "PD-L1" refers to programmed cell death ligand 1. As used herein, the term "human PD-L1" refers to the human PD-L1 protein encoded by the wild-type human PD-L1 gene (e.g., RefSeq accession number NM_014143.4). An exemplary immature amino acid sequence of human PD-L1 is provided in RefSeq accession number NP_054862.1.

[0046] As used herein, the term "antibody" includes full-length antibodies, antigen-binding fragments of full-length antibodies, and molecules comprising antibody CDR, VH region, and / or VL region. Examples of antibodies include, but are not limited to, monoclonal antibodies, recombinant antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetramer antibodies comprising two heavy chain molecules and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain-antibody heavy chain pairs, intracellular antibodies, heteroconjugated antibodies, antibody-drug conjugates, single-domain antibodies, monovalent antibodies, single-chain antibodies or single-chain Fv (scFv), camel-derived antibodies, affinity molecules, Fab fragments, F(ab')2 fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), and antigen-binding fragments of any of the above. In some embodiments, the antibodies described herein refer to a population of polyclonal antibodies. Antibodies can be any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In some embodiments, the antibody described herein is an IgG antibody, or its class (e.g., human IgG1 or IgG4) or subclass. In one specific embodiment, the antibody is a humanized antibody. In another specific embodiment, the antibody is a human antibody.

[0047] As used herein, the term “CDR” or “complementarity-determining region” refers to a discontinuous antigen-binding site found within the variable region of the heavy and light chain polypeptides. In some embodiments, the CDR of the antibodies disclosed herein is determined according to Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immune interest (1991), each of which is incorporated herein by reference in its entirety. In some embodiments, the CDR of the antibodies disclosed herein is determined according to the Chothia numbering scheme (see, for example, Chothia C and Lesk AM, (1987), J Mol Biol 196: 901-917; Al-Lazikani B et al., (1997) J Mol Biol 273: 927-948; Chothia C et al., (1992) J Mol Biol 227: 799-817; Tramontano A et al., (1990) J Mol Biol 215(1): 175-82; and U.S. Patent No. 7,709,226, all of which are incorporated herein by reference in their entirety). In some embodiments, the CDR of the antibodies disclosed herein is determined according to MacCallum RM et al., (1996) J Mol Biol 262: 732-745, which is incorporated herein by reference in its entirety. See also, for example, Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, edited by Kontermann and Dribel, Chapter 31, pp. 422–439, Springer-Verlag, Berlin (2001), which is incorporated herein by reference in its entirety. In some embodiments, the CDR of the antibodies disclosed herein is determined according to the IMGT numbering system as described in the following references: Lefranc MP, (1999) The Immunologist 7: 132–136; Lefranc MP et al., (1999) Nucleic Acids Res 27: 209–212, each of which is incorporated herein by reference in its entirety; and Lefranc MP et al., (2009) Nucleic Acids Res 37: D1006–D1012.In some embodiments, the CDRs of the antibodies disclosed herein are determined according to the AbM numbering scheme, which relates to the AbM hypervariable region, represents a compromise between the Kabat CDR and the Chothia structural loop, and is used by the AbM antibody modeling software of Oxford Molecular (Oxford Molecular Group, Inc.), which is incorporated herein by reference in its entirety. In some embodiments, the CDRs of the antibodies disclosed herein are determined according to the AHo numbering system, as described in Honegger and Pluckthun, J. Mol. Biol. 309:657-670 (2001), which is incorporated herein by reference in its entirety. In some embodiments, the CDRs of the antibodies disclosed herein are each determined independently according to one of the Kabat, Chothia, MacCallum, IMGT, AHo, or AbM numbering schemes or by structural analysis of a multispecific molecule, wherein the structural analysis identifies residues in the variable region predicted to contact the epitope. CDRH1, CDRH2, and CDRH3 represent heavy chain CDRs, and CDRL1, CDRL2, and CDRL3 represent light chain CDRs.

[0048] As used herein, the terms “VH” and “VL” refer to the variable regions of the antibody heavy and light chains, respectively, as described in Kabat et al., (1991) Sequences of Proteins of Immunological Interest (NIH Publication No. 91-3242, Bethesda), which is incorporated herein by reference in its entirety.

[0049] As used herein, the term "constant region" is common in the art. A constant region is a portion of an antibody, such as the carboxyl-terminal portion of the light chain and / or heavy chain, that does not directly participate in the binding of the antibody to the antigen but can exhibit various effector functions, such as interaction with Fc receptors (e.g., Fcγ receptors).

[0050] As used herein, the term “heavy chain” when referring to antibody use can refer to any different type of amino acid sequence based on the constant region, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which produce antibodies of the IgA, IgD, IgE, IgG, and IgM classes, including subclasses of IgG such as IgG1, IgG2, IgG3, and IgG4.

[0051] As used herein, the term "light chain" when referring to antibody use can refer to any different type based on the amino acid sequence of the constant region, such as kappa (κ) or lambda (λ). Light chain amino acid sequences are well known in the art. In this particular embodiment, the light chain is a human light chain.

[0052] As used herein, the term “EU numbering system” refers to the EU numbering convention for antibody constant regions, as described in Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat et al., Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services, 5th edition, 1991.

[0053] As used herein, the term "specific binding" refers to the specificity of an antibody to an antigen, as understood by those skilled in the art. Binding molecules that specifically bind to an antigen typically have a molecular weight of less than 1 × 10⁻⁶. -6 The equilibrium dissociation constant (KD) of M is bound to the antigen, as measured by, for example, an ELISA assay, surface plasmon resonance, or other suitable assays known in the art. Those skilled in the art will understand that in some embodiments, the binding molecule can specifically bind to different antigens (e.g., different antigens sharing a common epitope recognized by the binding molecule).

[0054] As used herein, in the context of Fc, the term "fucosylated-free" refers to the substantial absence of fucose that is directly or indirectly covalently attached to residue 297 of the Fc region of human IgG1 or to a corresponding residue in non-IgG1 or non-human IgG1 immunoglobulin, as designated by the EU numbering system. Therefore, in compositions comprising multiple fucosylated antibodies, at least 70% of the antibody will not be directly or indirectly (e.g., via an inserted sugar) fucosylated at residue 297 of the Fc region of the antibody, and in some embodiments, at least 80%, 85%, 90%, 95%, or 99% will not be directly or indirectly fucosylated at residue 297 of the Fc region.

[0055] As used in this article, the term "CTLA-4 inhibitor" refers to a molecule that can inhibit the binding of CTLA-4 to its ligand, differentiation cluster 80 (CD80).

[0056] As used in this article, the term "PD-1 inhibitor" refers to a molecule that can inhibit the binding of PD-1 to its ligand, programmed death ligand 1 (PD-L1).

[0057] As used herein, "epitope" is a term in the art and refers to a localized region of an antigen to which an antibody can specifically bind. An epitope can be, for example, a continuous amino acid of a polypeptide (linear or continuous epitope), or an epitope can be, for example, derived from two or more discontinuous regions of one or more polypeptides (i.e., conformational, nonlinear, discontinuous, or non-continuous epitopes). In some embodiments, the epitope to which the antibody binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography, ELISA assays, hydrogen / deuterium exchange binding mass spectrometry (e.g., liquid chromatography-electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, crystallization can be achieved using any method known in the art (e.g., Giegé R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; Chayen NE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303). Antibody-antigen crystals can be studied using X-ray diffraction techniques well known in the art, and can be refined using computer software such as X-PLOR (Yale University, 1992, published by Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985), vols. 114 and 115, edited by Wyckoff HW et al.; US 2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, edited by Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323). Mutagenesis mapping studies can be performed using any method known to those skilled in the art. See, for example, Champe M et al., (1995) JBiol Chem 270: 1388-1394 and Cunningham BC and Wells JA (1989) Science 244: 1081-1085 for information on mutagenesis techniques, including alanine scanning mutagenesis.In one specific implementation, alanine scanning mutagenesis is used to determine the epitopes of the antibody.

[0058] As used herein, the terms “treat,” “treating,” and “treatment” refer to the therapeutic or preventative measures described herein. A “treatment” method includes administering antibodies to a subject who has a disease or condition, or is predisposed to developing such a disease or condition, in order to prevent, cure, delay, reduce the severity of the disease or condition or a recurrent disease or condition, or improve one or more symptoms of the disease or condition or a recurrent disease or condition, or in order to prolong the subject’s expected survival in the absence of such treatment.

[0059] As used herein, the term "treatment combination" refers to a combination of a first therapy and a second therapy administered to a subject. The first therapy and the second therapy may be administered simultaneously (in the same pharmaceutical composition or in separate pharmaceutical compositions) or sequentially in any order.

[0060] As used herein, in the context of administering a therapy to a subject, the term "effective amount" refers to the amount of therapy that achieves the desired preventive or therapeutic effect. An effective amount of a combination of first and second therapies includes a first amount of the first therapy and a second amount of the second therapy, wherein the administration of this combination of therapies achieves the desired preventive or therapeutic effect.

[0061] As used herein, the term "subject" includes any human or non-human animal. In some embodiments, the subject is a human or a non-human mammal. In some embodiments, the subject is a human.

[0062] The determination of the "percentage of identity" between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be accomplished using mathematical algorithms. A specific, non-limiting example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin S and Altschul SF (1990) PNAS 87: 2264-2268, modified by Karlin S and Altschul SF (1993) PNAS 90: 5873-5877, each of which is incorporated herein by reference in its entirety. This algorithm is also incorporated in the NBLAST and XBLAST procedures of Altschul SF et al., (1990) J Mol Biol 215: 403, which is incorporated herein by reference in its entirety. BLAST nucleotide searches can be performed using the NBLAST nucleotide procedure parameter set, for example, setting score = 100 and word length = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST program parameter set, for example, with a score of 50 and a word length of 3, to obtain amino acid sequences homologous to the protein molecules described herein. For gapped alignments for comparative purposes, Gapped BLAST, as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, is used, which is incorporated herein by reference in its entirety. Alternatively, iterative searches can be performed using PSI BLAST to detect distant relationships between molecules (ibid.). When using BLAST, Gapped BLAST, and PSI BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) website on the World Wide Web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is incorporated herein by reference in its entirety. This algorithm was incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When comparing amino acid sequences using the ALIGN program, a PAM120 weighted residue table, a 12-bit gap length penalty, and a 4-bit gap penalty can be used. The percentage of identity between two sequences can be determined using techniques similar to those described above, regardless of whether gaps are allowed. When calculating the percentage of identity, only exact matches are typically counted.

[0063] As used herein, the term “Eastern Oncology Cooperative Group Performance Status” refers to the level on the Eastern Oncology Cooperative Group Performance Status Scale determined for the subject prior to treatment. The Eastern Oncology Cooperative Group Performance Status Scale is well-known in the field and describes a patient’s functional level in self-care, daily activities, and physical abilities (walking, working, etc.).

[0064] As used herein, the term “Cockcroft-Gault method” refers to a method for calculating creatinine clearance (CrCl) from a subject’s serum creatinine, age, and weight. This method is well known in the art and is described in Cockcroft DW, Gault MH. Nephron. 1976; 16(1):31-41), which is incorporated herein by reference in its entirety.

[0065] As used herein, the terms “Records for Evaluation of Treatment of Solid Tumors Version 1.1” and “RECIST 1.1” refer to a standardized radiological imaging method used to measure the response of a tumor to treatment. RECIST 1.1 is well known in the art and is described, for example, in Therasse P et al., JNCI. 2000;92(3):205-216.

[0066] Anti-ILT2 antibody Anti-human ILT2 antibodies that can be used in the methods and uses described herein include, but are not limited to, those described below.

[0067] In some embodiments, the antibody that specifically binds to human ILT2 comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7. In some embodiments, the antibody that specifically binds to human ILT2 comprises: a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8.

[0068] In some embodiments, the antibody that specifically binds to human ILT2 comprises the amino acid sequences CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO: 1, 2, and 3, respectively. In some embodiments, the antibody that specifically binds to human ILT2 comprises the amino acid sequences CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 4, 5, and 6, respectively. In some embodiments, the antibody comprises the amino acid sequences CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 1, 2, 3, 4, 5, and 6, respectively.

[0069] In some embodiments, the antibody that specifically binds to human ILT2 comprises a VH containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 7. In some embodiments, the antibody comprises a VH containing the amino acid sequence shown in SEQ ID NO: 7.

[0070] In some embodiments, the antibody that specifically binds to human ILT2 comprises a VL containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the antibody comprises a VL containing the amino acid sequence shown in SEQ ID NO: 8.

[0071] In some embodiments, the antibody that specifically binds to human ILT2 comprises: VH, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 7; and VL, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the antibody comprises: VH, which comprises the amino acid sequence shown in SEQ ID NO: 7; and VL, which comprises the amino acid sequence shown in SEQ ID NO: 8.

[0072] In some embodiments, the antibody that specifically binds to human ILT2 comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In some embodiments, the heavy chain constant region is IgG4. In some embodiments, the antibody comprises a light chain constant region selected from the group consisting of the human κ light chain constant region and the human λ light chain constant region.

[0073] In some embodiments, the antibody that specifically binds to human ILT2 comprises a heavy chain constant region (e.g., the IgG4 constant region) or a fragment thereof, which contains an S228P mutation numbered according to the EU numbering system. In some embodiments, the heavy chain constant region is the human IgG4 heavy chain constant region, which contains proline at position 228, numbered according to the EU numbering system.

[0074] In some embodiments, the antibody that specifically binds to human ILT2 comprises a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 9. In some embodiments, the antibody comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 9.

[0075] In some embodiments, the antibody that specifically binds to human ILT2 comprises a light chain having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the antibody comprises a light chain having the amino acid sequence shown in SEQ ID NO: 10.

[0076] In some embodiments, the antibody that specifically binds to human ILT2 comprises: a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 9; and a light chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the antibody comprises: a heavy chain containing the amino acid sequence shown in SEQ ID NO: 9; and a light chain containing the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 9, and the amino acid sequence of the light chain consists of the amino acid sequence shown in SEQ ID NO: 10.

[0077] In some implementations, the antibody that specifically binds to human ILT2 is AGEN1571.

[0078] The amino acid sequences of exemplary anti-ILT2 antibodies are provided in Table 1 of this document.

[0079] Table 1. Amino acid sequences of exemplary anti-ILT2 antibodies .

[0080]

[0081] PD-1 inhibitors Human PD-1 inhibitors that can be used in the methods and uses described herein include, but are not limited to, those described below.

[0082] In some embodiments, the human PD-1 inhibitor is an antibody that specifically binds to human PD-1 or human PD-L1. In some embodiments, the antibody that specifically binds to human PD-1 comprises: a heavy chain variable region (VH) containing the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 17. In some embodiments, the antibody that specifically binds to human PD-1 comprises: a light chain variable region (VL) containing the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the antibody that specifically binds to human PD-1 comprises: a heavy chain variable region (VH) containing the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 17; and a light chain variable region (VL) containing the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 18.

[0083] In some embodiments, the antibody that specifically binds to human PD-1 comprises the amino acid sequences CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO: 11, 12, and 13, respectively. In some embodiments, the antibody that specifically binds to human PD-1 comprises the amino acid sequences CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 14, 15, and 16, respectively. In some embodiments, the antibody that specifically binds to human PD-1 comprises the amino acid sequences CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 11, 12, 13, 14, 15, and 16, respectively.

[0084] In some embodiments, the antibody that specifically binds to human PD-1 comprises a VH containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 17. In some embodiments, the antibody that specifically binds to human PD-1 comprises a VH containing the amino acid sequence shown in SEQ ID NO: 17.

[0085] In some embodiments, the antibody that specifically binds to human PD-1 comprises a VL containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the antibody that specifically binds to human PD-1 comprises a VL containing the amino acid sequence shown in SEQ ID NO: 18.

[0086] In some embodiments, the antibody that specifically binds to human PD-1 comprises: VH, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 17; and VL, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the antibody that specifically binds to human PD-1 comprises: VH, which comprises the amino acid sequence shown in SEQ ID NO: 17; and VL, which comprises the amino acid sequence shown in SEQ ID NO: 18.

[0087] In some embodiments, the antibody that specifically binds to human PD-1 comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In some embodiments, the heavy chain constant region is IgG1. In some embodiments, the heavy chain constant region is IgG2. In some embodiments, the antibody comprises a light chain constant region selected from the group consisting of the human κ light chain constant region and the human λ light chain constant region.

[0088] In some embodiments, the antibody that specifically binds to human PD-1 comprises the IgG4 heavy chain constant region. In some embodiments, the heavy chain constant region is the human IgG4 heavy chain constant region, which contains proline at position 228 according to the EU numbering system.

[0089] In some embodiments, the antibody that specifically binds to human PD-1 comprises a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 19. In some embodiments, the antibody comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 19.

[0090] In some embodiments, the antibody that specifically binds to human PD-1 comprises a light chain having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 20. In some embodiments, the antibody comprises a light chain having the amino acid sequence shown in SEQ ID NO: 20.

[0091] In some embodiments, the antibody that specifically binds to human PD-1 comprises: a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 19; and a light chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 20. In some embodiments, the antibody that specifically binds to human PD-1 comprises: a heavy chain containing the amino acid sequence shown in SEQ ID NO: 19; and a light chain containing the amino acid sequence shown in SEQ ID NO: 20. In some embodiments, the amino acid sequence of the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 19, and the amino acid sequence of the light chain consists of the amino acid sequence shown in SEQ ID NO: 20.

[0092] In some implementations, the antibody that specifically binds to human PD-1 is batitimab.

[0093] The amino acid sequences of exemplary anti-PD-1 antibodies are provided in Table 2 of this document.

[0094] Table 2. Amino acid sequences of exemplary anti-PD-1 antibodies .

[0095]

[0096] In some implementations, the antibodies that specifically bind to human PD-1 or human PD-L1 are adebelimumab, atezolizumab, avelumumab, camrelizumab, simipremab, cochilimumab, dostalimumab, durvalumab, enronsubab, envorizumab, nivolumab, pembrolizumab, pendilizumab, palolizumab, pectilizumab, pravalimumab, slulizumab, sintilimumab, socazolimumab, sugelizumab, tagorizumab, tislelizumab, toripalimab, and cepalimumab.

[0097] Other non-limiting examples of anti-PD-1 antibodies that can be used in the treatments described herein are disclosed in the following patents and patent applications, which are incorporated herein by reference in their entirety for all purposes: U.S. Patent No. 6,808,710; U.S. Patent No. 7,332,582; U.S. Patent No. 7,488,802; U.S. Patent No. 8,008,449; U.S. Patent No. 8,114,845; U.S. Patent No. 8,168,757; U.S. Patent No. 8,354,509; U.S. Patent No. 8,686,119; U.S. Patent No. 8,735,553; U.S. Patent No. 8,747,847; U.S. Patent No. 8,779,105; U.S. Patent No. 8,927,697; U.S. Patent No. 8,993,731; U.S. Patent No. 9,102,727; U.S. Patent No. 9,205,148; U.S. Publication No. US 2013 / 0202623 A1; US ​​Publication No. US 2013 / 0291136 A1; US ​​Publication No. US 2014 / 0044738 A1; US ​​Publication No. US 2014 / 0356363 A1; US ​​Publication No. US 2016 / 0075783A1; and PCT Publication No. WO 2013 / 033091 A1; PCT Publication No. WO 2015 / 036394 A1; PCT Publication No. WO2014 / 179664 A2; PCT Publication No. WO 2014 / 209804 A1; PCT Publication No. WO 2014 / 206107 A1; PCT Publication No. WO 2015 / 058573 A1; PCT Publication No. WO 2015 / 085847 A1; PCT Publication No. WO 2015 / 200119 A1; PCT Publication No. WO 2016 / 015685 A1; and PCT Publication No. WO 2016 / 020856 A1.

[0098] Other examples of anti-PD-L1 antibodies that can be used in the treatments described herein are disclosed in the following patents and patent applications, which are incorporated herein by reference in their entirety for all purposes: U.S. Patent No. 7,943,743; U.S. Patent No. 8,168,179; U.S. Patent No. 8,217,149; U.S. Patent No. 8,552,154; U.S. Patent No. 8,779,108; U.S. Patent No. 8,981,063; U.S. Patent No. 9,175,082; U.S. Publication No. US2010 / 0203056 A1; U.S. Publication No. US2003 / 0232323 A1; U.S. Publication No. US2013 / 0323249 A1; U.S. Publication No. US2014 / 0341917 A1; U.S. Publication No. US2014 / 0044738 A1; U.S. Publication No. US2015 / 0203580 A1; US ​​Publication No. US 2015 / 0225483 A1; US ​​Publication No. US 2015 / 0346208 A1; US ​​Publication No. US 2015 / 0355184 A1; and PCT Publication No. WO 2014 / 100079 A1; PCT Publication No. WO 2014 / 022758A1; PCT Publication No. WO 2014 / 055897 A2; PCT Publication No. WO 2015 / 061668 A1; PCT Publication No. WO 2015 / 109124 A1; PCT Publication No. WO 2015 / 195163 A1; PCT Publication No. WO 2016 / 000619 A1; and PCT Publication No. WO2016 / 030350 A1.

[0099] In some implementations, the human PD-1 inhibitor is pildizumab.

[0100] CTLA-4 inhibitors Human CTLA-4 inhibitors that can be used in the methods and uses described herein include, but are not limited to, those described below.

[0101] In some embodiments, the human CTLA-4 inhibitor is an antibody that specifically binds to human CTLA-4. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises: a heavy chain variable region (VH) containing the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 27. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises: a light chain variable region (VL) containing the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 28. In some embodiments, the antibody comprises: a heavy chain variable region (VH) containing the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 27; and a light chain variable region (VL) containing the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 28.

[0102] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises the amino acid sequences CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO: 21, 22, and 23, respectively. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises the amino acid sequences CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 24, 25, and 26, respectively. In some embodiments, the antibody comprises the amino acid sequences CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO: 21, 22, 23, 24, 25, and 26, respectively.

[0103] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a VH containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 27. In some embodiments, the antibody comprises a VH containing the amino acid sequence shown in SEQ ID NO: 27.

[0104] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a VL containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 28. In some embodiments, the antibody comprises a VL containing the amino acid sequence shown in SEQ ID NO: 28.

[0105] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises: VH, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 27; and VL, which comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 28. In some embodiments, the antibody comprises: VH, which comprises the amino acid sequence shown in SEQ ID NO: 27; and VL, which comprises the amino acid sequence shown in SEQ ID NO: 28.

[0106] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In some embodiments, the heavy chain constant region is IgG1. In some embodiments, the heavy chain constant region is IgG2. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises the IgG1 heavy chain constant region.

[0107] In some embodiments, the antibody comprises a light chain constant region selected from the group consisting of the human κ light chain constant region and the human λ light chain constant region.

[0108] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a variant of the human IgG heavy chain constant region as a wild-type human IgG heavy chain constant region, wherein the variant human IgG heavy chain constant region binds to FcγRIIIA with a higher affinity than the wild-type human IgG heavy chain constant region.

[0109] In some implementations, the IgG region of an antibody that specifically binds to human CTLA-4 has increased affinity for FcγRIIIA, for example, compared to antibodies with a wild-type Fc region, such as IgG1Fc. Sequence alterations leading to increased affinity for FcγRIIIA are, for example, described in Kellner et al. Methods 65: 105-113 (2014), Lazar et al. Proc Natl Acad Sci 103: 4005-4010 (2006), Shields et al. J Biol Chem The references in .276(9): 6591-6604(2001) are known in the art, and each of these references is incorporated herein by reference in its entirety.

[0110] In some implementations, the antibody that specifically binds to human CTLA-4 is unfucosylated.

[0111] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, which contains mutations selected from the group consisting of: L235V, G236A, S239D, F243L, T256A, K290A, R292P, S298A, Y300L, V305I, A330L, I332E, E333A, K334A, A339T, and P396L, and combinations thereof, numbered according to the EU numbering system.

[0112] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising S239D according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising T256A according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising K290A according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising S298A according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising I332E according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising E333A according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising A339T according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, the heavy chain constant region or fragment thereof comprising S239D and I332E according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises an IgG1 heavy chain constant region containing the S239D / I332E mutation numbered according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, which contains S239D, A330L, and I332E mutations numbered according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises an IgG1 heavy chain constant region containing the S239D / A330L / I332E mutation numbered according to the EU numbering system.In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, which comprises S298A, E333A, and K334A according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, which comprises G236A, S239D, and I332E according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain constant region (e.g., the IgG1 constant region) or a fragment thereof, which comprises F243L, R292P, Y300L, V305I, and P396L according to the EU numbering system. In some embodiments, the antibody that specifically binds to human CTLA-4 comprises an IgG1 heavy chain constant region containing the L235V / F243L / R292P / Y300L / P396L mutation, numbered according to the EU numbering system. In some embodiments, the heavy chain constant region is the human IgG1 heavy chain constant region, which contains aspartic acid at position 239, leucine at position 330, and glutamic acid at position 332, numbered according to the EU numbering system.

[0113] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 29. In some embodiments, the antibody comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 29.

[0114] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises a light chain having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 30. In some embodiments, the antibody comprises a light chain having the amino acid sequence shown in SEQ ID NO: 30.

[0115] In some embodiments, the antibody that specifically binds to human CTLA-4 comprises: a heavy chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 29; and a light chain containing an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 30. In some embodiments, the antibody comprises: a heavy chain containing the amino acid sequence shown in SEQ ID NO: 29; and a light chain containing the amino acid sequence shown in SEQ ID NO: 30. In some embodiments, the amino acid sequence of the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 29, and the amino acid sequence of the light chain consists of the amino acid sequence shown in SEQ ID NO: 30.

[0116] In some embodiments, the antibody that specifically binds to human CTLA-4 is selected from the group consisting of: botelimab, ipilimumab, and trimemumab. In some embodiments, the antibody that specifically binds to human CTLA-4 is botelimab.

[0117] The amino acid sequences of exemplary anti-CTLA-4 antibodies are provided in Table 3 of this document.

[0118] Table 3. Amino acid sequences of exemplary anti-CTLA-4 antibodies .

[0119]

[0120] Methods of treating cancer This disclosure provides methods for treating cancer using an antibody that specifically binds to human ILT2, and a combination of the antibody with a human CTLA-4 inhibitor (e.g., an antibody that specifically binds to human CTLA-4) and a human PD-1 inhibitor (e.g., an antibody that specifically binds to human PD-1).

[0121] This disclosure also provides methods for treating cancer using antibodies that specifically bind to human ILT2, and combinations of such antibodies with checkpoint targets (e.g., CTLA-4 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, or PD-1 inhibitors). In some embodiments, the checkpoint target comprises an antagonistic anti-CTLA-4 antibody, an antagonistic anti-PD-L1 antibody, an antagonistic anti-PD-L2 antibody, or an antagonistic anti-PD-1 antibody. In some embodiments, the checkpoint target comprises a small molecule (e.g., inhibitory) targeting CTLA-4, PD-L1 antibody, PD-L2, or PD-1. In some embodiments, the checkpoint target comprises a peptide (e.g., inhibitory) targeting CTLA-4, PD-L1 antibody, PD-L2, or PD-1. In some embodiments, the checkpoint target comprises an Fc fusion protein (e.g., an inhibitory IgG-Fc-labeled protein) targeting CTLA-4, PD-L1 antibody, PD-L2, or PD-1.

[0122] This disclosure also provides methods for enhancing the immune response of a subject in need by using an antibody that specifically binds to human ILT2, and by combining such an antibody with a checkpoint target (e.g., a CTLA-4 inhibitor, a PD-L1 inhibitor, a PD-L2 inhibitor, or a PD-1 inhibitor). In some embodiments, the methods disclosed herein for enhancing the immune response of a subject in need include using an antibody that specifically binds to human ILT2, and by combining such an antibody with a human CTLA-4 inhibitor (e.g., an antibody that specifically binds to human CTLA-4) and / or a human PD-1 inhibitor (e.g., an antibody that specifically binds to human PD-1). In some embodiments, the enhanced immune response includes increased CD25 expression on T cells, NK cells, and / or NKT cells compared to a reference or control. In some embodiments, the enhanced immune response includes increased secretion of TNFα, IFNγ, and / or IL-1β cytokines compared to a reference or control. In some embodiments, the T cells are CD8+ cells. + T cells. In some implementations, NK cells are CD3+ cells. - / CD56 + NK cells. In some implementations, NKT cells are CD3+ cells. + / CD56 +NKT cells. In some embodiments, the enhanced immune response includes enhanced FcγR-mediated immune activation compared to a reference or control. In some embodiments, FcγR-mediated immune activation includes FcγRIIIA receptor (CD16) signaling. In some embodiments, the reference or control is a published or historical reference of the subject's immune response to an ILT2 inhibitor. In some embodiments, the reference or control is a published or historical reference of the subject's immune response to a CTLA-4 inhibitor. In some embodiments, the reference or control is a published or historical reference of the subject's immune response to a PD-1 inhibitor.

[0123] In one aspect, this disclosure provides a method for treating cancer in a subject of need, the method comprising administering to the subject a first antibody that specifically binds to human ILT2, the first antibody being in a dose of about 1 mg to about 2000 mg, wherein the first antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8.

[0124] In another aspect, this article provides a method for treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, the VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; VL, the VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human PD-1 inhibitor.

[0125] In another aspect, this article provides a method for treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, the VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; VL, the VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and a human CTLA-4 inhibitor.

[0126] In another aspect, this article provides a method for treating cancer in a subject of need, the method comprising administering to the subject: a first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, the VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; VL, the VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; a human PD-1 inhibitor; and a human CTLA-4 inhibitor.

[0127] The methods described herein may include administering a first antibody that specifically binds to human ILT2 as a single therapy. In some embodiments, the methods described herein include any combination of the first antibody described herein that specifically binds to human ILT2 with the human PD-1 inhibitor and / or the human CTLA-4 inhibitor described herein.

[0128] In some embodiments, the method includes administering a first antibody that specifically binds to human ILT2, and a human PD-1 inhibitor as a second antibody that specifically binds to human PD-1 or human PD-L1. In some embodiments, the second antibody specifically binds to human PD-1. In some embodiments, the method includes administering a first antibody that specifically binds to human ILT2, and a second antibody comprising: VH, which comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 17; and VL, which comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the second antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences shown in SEQ ID NO: 11, 12, 13, 14, 15, and 16, respectively. In some embodiments, the second antibody comprises the VH amino acid sequence of SEQ ID NO: 17, and / or the VL amino acid sequence of SEQ ID NO: 18. In some embodiments, the second antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 17 and 18, respectively. In some embodiments, the second antibody comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM. In some embodiments, the second antibody comprises a heavy chain constant region, which is the human IgG4 heavy chain constant region, containing proline at position 228 according to the EU numbering system. In some embodiments, the second antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 19, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the second antibody comprises a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 19 and 20, respectively. In some embodiments, the secondary antibody is selected from the group consisting of: batitilimab, adebelimumab, atezolizumab, avelumumab, camrelizumab, cimiprizumab, cochilimumab, dostalimumab, durvalumab, emronsubab, envorimab, nivolumab, pembrolizumab, pendilizumab, palolizumab, pectilimab, prazolimumab, slulizumab, sintilimumab, socarzolimumab, sugalizumab, tagorizumab, tislelizumab, toripalimab, and cepalimumab. In some embodiments, the secondary antibody is batitilimab.

[0129] In some embodiments, the method includes administering a first antibody that specifically binds to human ILT2, and a human CTLA-4 inhibitor as a third antibody that specifically binds to human CTLA-4. In some embodiments, the method includes administering a first antibody that specifically binds to human ILT2, and a third antibody comprising: VH, which comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 27; and VL, which comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 28. In some embodiments, the third antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences shown in SEQ ID NO: 21, 22, 23, 24, 25, and 26, respectively. In some embodiments, the third antibody comprises the VH amino acid sequence of SEQ ID NO: 27, and / or the VL amino acid sequence of SEQ ID NO: 28. In some embodiments, the third antibody comprises the VH amino acid sequence and the VL amino acid sequence, respectively, of SEQ ID NO: 27 and 28. In some embodiments, the third antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, ... 4、 IgA1, IgA2, and IgM. In some embodiments, the third antibody comprises a heavy chain constant region, which is the human IgG1 heavy chain constant region, comprising aspartic acid at position 239, leucine at position 330, and glutamic acid at position 332 according to the EU numbering system. In some embodiments, the third antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 30. In some embodiments, the third antibody comprises a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 29 and 30, respectively. In some embodiments, the third antibody is selected from the group consisting of: botelimab, ipilimumab, and trimemumab. In some embodiments, the third antibody is botelimab.

[0130] In some embodiments, the method includes administering a first antibody that specifically binds to human ILT2, a second antibody that specifically binds to human PD-1, and a third antibody that specifically binds to human CTLA-4.

[0131] In some embodiments, the first antibody is in the form of about 1 mg to about 2000 mg, about 1 mg to about 1000 mg, about 3 mg to about 300 mg, about 10 mg to about 100 mg, about 1 mg to about 600 mg, about 1 mg to about 300 mg, about 1 mg to about 100 mg, about 1 mg to about 30 mg, about 1 mg to about 10 mg, about 1 mg to about 3 mg, about 10 mg to about 2000 mg, about 10 mg to about 1000 mg, or about 10 mg to about 600 mg. Administered in doses of g, about 10 mg to about 300 mg, about 10 mg to about 30 mg, about 30 mg to about 1000 mg, about 30 mg to about 600 mg, about 30 mg to about 300 mg, about 30 mg to about 100 mg, about 100 mg to about 1000 mg, about 100 mg to about 600 mg, about 100 mg to about 300 mg, about 300 mg to about 2000 mg, about 300 mg to about 1000 mg, or about 300 mg to about 600 mg.

[0132] In some embodiments, the first antibody is administered at doses of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1250 mg, about 1500 mg, about 1750 mg, or about 2000 mg.

[0133] In some embodiments, the primary antibody is administered weekly. In some embodiments, the primary antibody is administered every two weeks. In some embodiments, the primary antibody is administered every three weeks. In some embodiments, the primary antibody is administered every four weeks. In some embodiments, the primary antibody is administered every five weeks. In some embodiments, the primary antibody is administered every six weeks.

[0134] In some embodiments, the primary antibody is administered at a dose of about 1 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 3 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 10 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 30 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 100 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 300 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 1000 mg every 3 weeks. In some embodiments, the primary antibody is administered at a dose of about 1 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of about 3 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of about 10 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of about 30 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of about 100 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of approximately 300 mg every 6 weeks. In some embodiments, the primary antibody is administered at a dose of approximately 1000 mg every 6 weeks.

[0135] In some implementations, human PD-1 inhibitors are administered at doses of about 1 mg to 1000 mg, 100 mg to 750 mg, 100 mg to 500 mg, 100 mg to 450 mg, 150 mg to 750 mg, 150 mg to 500 mg, 150 mg to 450 mg, 200 mg to 750 mg, 200 mg to 500 mg, 200 mg to 450 mg, 300 mg to 750 mg, 300 mg to 500 mg, 300 mg to 450 mg, 400 mg to 750 mg, 400 mg to 500 mg, 400 mg to 450 mg, 450 mg to 750 mg, or 450 mg to 500 mg.

[0136] In some embodiments, the human PD-1 inhibitor is administered at a dose of about 100 mg, about 120 mg, about 140 mg, about 160 mg, about 180 mg, about 200 mg, about 220 mg, about 240 mg, about 260 mg, about 280 mg, about 300 mg, about 320 mg, about 340 mg, about 360 mg, about 380 mg, about 400 mg, about 420 mg, about 440 mg, about 450 mg, about 460 mg, about 480 mg, about 500 mg, about 520 mg, about 540 mg, about 560 mg, about 580 mg, about 600 mg, about 620 mg, about 640 mg, about 660 mg, about 680 mg, about 700 mg, about 720 mg, or about 740 mg. In some embodiments, the human PD-1 inhibitor is administered at a dose of about 240 mg. In some implementations, human PD-1 inhibitors are administered at a dose of approximately 450 mg.

[0137] In some implementations, human PD-1 inhibitors are administered at doses of about 0.01 mg / kg, about 0.03 mg / kg, about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 3 mg / kg, about 6 mg / kg, or about 10 mg / kg.

[0138] In some embodiments, the human PD-1 inhibitor is administered once weekly. In some embodiments, the human PD-1 inhibitor is administered once every two weeks. In some embodiments, the human PD-1 inhibitor is administered once every three weeks. In some embodiments, the human PD-1 inhibitor is administered once every four weeks. In some embodiments, the human PD-1 inhibitor is administered once every five weeks. In some embodiments, the human PD-1 inhibitor is administered once every six weeks. In some embodiments, the human PD-1 inhibitor is administered at a dose of 450 mg every three weeks. In some embodiments, the human PD-1 inhibitor is administered at a dose of 3 mg / kg every three weeks.

[0139] In some embodiments, human CTLA-4 inhibitors are administered at doses of approximately 1 mg to 1000 mg, 10 mg to 750 mg, 10 mg to 500 mg, 10 mg to 250 mg, 10 mg to 200 mg, 10 mg to 150 mg, 10 mg to 100 mg, 10 mg to 75 mg, 10 mg to 50 mg, 10 mg to 25 mg, 25 mg to 750 mg, 25 mg to 500 mg, 25 mg to 250 mg, and 25 mg. Administered at doses of up to 150 mg, 25 mg to 100 mg, 25 mg to 75 mg, 25 mg to 50 mg, 50 mg to 750 mg, 50 mg to 500 mg, 50 mg to 250 mg, 50 mg to 150 mg, 50 mg to 75 mg, 75 mg to 250 mg, 75 mg to 150 mg, 75 mg to 100 mg, 100 mg to 250 mg, 100 mg to 150 mg, or 150 mg to 250 mg.

[0140] In some embodiments, the human CTLA-4 inhibitor is administered at a dose of about 10 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, or about 250 mg. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of about 150 mg. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of about 75 mg. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of about 50 mg.

[0141] In some implementations, human CTLA-4 inhibitors are administered at doses of about 0.01 mg / kg, about 0.02 mg / kg, about 0.04 mg / kg, about 0.2 mg / kg, about 0.4 mg / kg, about 1 mg / kg, about 2 mg / kg, about 4 mg / kg, or about 6 mg / kg.

[0142] In some embodiments, the human CTLA-4 inhibitor is administered once weekly. In some embodiments, the human CTLA-4 inhibitor is administered every two weeks. In some embodiments, the human CTLA-4 inhibitor is administered every three weeks. In some embodiments, the human CTLA-4 inhibitor is administered every four weeks. In some embodiments, the human CTLA-4 inhibitor is administered every five weeks. In some embodiments, the human CTLA-4 inhibitor is administered every six weeks. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of 150 mg every six weeks. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of 2 mg / kg every six weeks. In some embodiments, the human CTLA-4 inhibitor is administered at a dose of 4 mg / kg every six weeks.

[0143] In some embodiments, the first antibody is administered intravenously. In some embodiments, the first antibody is administered intratumorally. In some embodiments, the first antibody is administered peritumorally. In some embodiments, a human PD-1 inhibitor or a human CTLA-4 inhibitor is administered intravenously. In some embodiments, a human PD-1 inhibitor or a human CTLA-4 inhibitor is administered intratumorally. In some embodiments, a human PD-1 inhibitor or a human CTLA-4 inhibitor is administered peritumorally.

[0144] In some embodiments, the primary antibody and the human PD-1 inhibitor are administered intravenously. In some embodiments, the primary antibody and the human PD-1 inhibitor are administered intratumorally. In some embodiments, the primary antibody and the human PD-1 inhibitor are administered peritumorally. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are administered intravenously. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are administered intratumorally. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are administered peritumorally. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are administered intravenously. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are administered intratumorally. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are administered peritumorally.

[0145] In some embodiments, the primary antibody, human PD-1 inhibitor, or human CTLA-4 inhibitor is administered via intravenous infusion over approximately 30 minutes. In some embodiments, the primary antibody, human PD-1 inhibitor, or human CTLA-4 inhibitor is administered via intravenous infusion over approximately 45 minutes. In some embodiments, the primary antibody, PD-1 inhibitor, or human CTLA-4 inhibitor is administered via intravenous infusion over approximately 60 minutes. In some embodiments, the primary antibody, human PD-1 inhibitor, or human CTLA-4 inhibitor is administered via intravenous infusion over approximately 90 minutes.

[0146] In some embodiments, the primary antibody and the human PD-1 inhibitor are each administered via intravenous infusion over approximately 30 minutes. In some embodiments, the primary antibody and the human PD-1 inhibitor are each administered via intravenous infusion over approximately 45 minutes. In some embodiments, the primary antibody and the human PD-1 inhibitor are each administered via intravenous infusion over approximately 60 minutes. In some embodiments, the primary antibody and the human PD-1 inhibitor are each administered via intravenous infusion over approximately 90 minutes. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 30 minutes. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 45 minutes. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 60 minutes. In some embodiments, the primary antibody and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 90 minutes. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 30 minutes. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 45 minutes. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 60 minutes. In some embodiments, the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are each administered via intravenous infusion over approximately 90 minutes.

[0147] In some embodiments, the human PD-1 inhibitor is administered to the subject after administration of the primary antibody. In some embodiments, the human CTLA-4 inhibitor is administered to the subject after administration of the primary antibody. In some embodiments, the human CTLA-4 inhibitor is administered to the subject after administration of both the primary antibody and the human PD-1 inhibitor.

[0148] In some embodiments, the dose of the primary antibody and the dose of the human PD-1 inhibitor are administered on the same day. In some embodiments, the dose of the primary antibody and the dose of the human PD-1 inhibitor are administered simultaneously. In some embodiments, the dose of the primary antibody is administered before the dose of the human PD-1 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.). In some embodiments, the dose of the primary antibody is administered after the dose of the human PD-1 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.).

[0149] In some embodiments, the dose of the primary antibody and the dose of the human CTLA-4 inhibitor are administered on the same day. In some embodiments, the dose of the primary antibody and the dose of the human CTLA-4 inhibitor are administered simultaneously. In some embodiments, the dose of the primary antibody is administered before the dose of the human CTLA-4 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.). In some embodiments, the dose of the primary antibody is administered after the dose of the human CTLA-4 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.).

[0150] In some embodiments, the doses of the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are administered on the same day. In some embodiments, the doses of the primary antibody, the human PD-1 inhibitor, and the human CTLA-4 inhibitor are administered simultaneously. In some embodiments, the dose of the primary antibody is administered before the dose of the human PD-1 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.), which in turn is administered before the dose of the human CTLA-4 inhibitor (e.g., 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 1.5 days, 2 days, 3 days, etc.).

[0151] In some embodiments, the dose of the primary antibody is a therapeutically effective amount. In some embodiments, the dose of the primary antibody and the dose of the human PD-1 inhibitor are both therapeutically effective amounts. In some embodiments, the dose of the primary antibody and the dose of the human CTLA-4 inhibitor are both therapeutically effective amounts. In some embodiments, the dose of the primary antibody, the dose of the human PD-1 inhibitor, and the dose of the human CTLA-4 inhibitor are all therapeutically effective amounts.

[0152] The methods, antibodies, or inhibitors described herein can be used to treat any disease or condition in a subject that would benefit from suppression of ILT2, CTLA-4, and / or PD-1 function. The methods, antibodies, or inhibitors described herein are particularly suitable for suppressing immune system tolerance to tumors and can therefore be used as immunotherapy for cancer subjects. For example, in some embodiments, this disclosure provides a method of treating a subject with cancer, the method comprising administering to the subject an effective amount of the antibody and / or inhibitor as described herein.

[0153] In some embodiments, the cancer treated according to the methods described herein is metastatic or locally advanced cancer (e.g., solid tumor). In some embodiments, the cancer is treated as a first-line cancer therapy according to the methods described herein after diagnosis of metastatic or locally advanced tumor (e.g., within 1, 2, 3, 4, 5, or 6 days after diagnosis; within 1, 2, 3, 4, 6, 8, or 12 weeks after diagnosis; or within 1, 2, 3, 4, 6, 8, or 12 months after diagnosis). In some embodiments, the cancer is treated as a first-line cancer therapy according to the methods described herein after diagnosis of tumor progression (e.g., within 1, 2, 3, 4, 5, or 6 days after diagnosis of tumor progression; within 1, 2, 3, 4, 6, 8, or 12 weeks after diagnosis; or within 1, 2, 3, 4, 6, 8, or 12 months after diagnosis), even though the tumor has been previously treated with different cancer therapies, optionally wherein the methods described herein are provided as a second-line cancer therapy. In some embodiments, cancer is treated as a first-line cancer therapy according to the method described herein after a diagnosis of toxicity to a different cancer therapy (e.g., within 1, 2, 3, 4, 5, or 6 days after a diagnosis of toxicity to a different cancer therapy; within 1, 2, 3, 4, 6, 8, or 12 weeks after a diagnosis of toxicity to a different cancer therapy; or within 1, 2, 3, 4, 6, 8, or 12 months after a diagnosis of toxicity to a different cancer therapy), optionally wherein the method described herein is provided as a second-line cancer therapy. In some embodiments, the cancer treated according to the method described herein is a metastatic or locally advanced cancer (e.g., a solid tumor) for which no standard therapy is available. In other embodiments, the cancer treated according to the method described herein is a metastatic or locally advanced cancer (e.g., a solid tumor) for which standard therapy has failed (i.e., the cancer has progressed after standard therapy). In some embodiments, if the cancer is refractory to the therapy, it is considered a therapy failure. In some embodiments, if the cancer recurs after a complete or partial response to the therapy, it is considered a therapy failure. In some embodiments, the metastatic or locally advanced cancer (e.g., a solid tumor) has been confirmed histologically or cytologically.

[0154] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer (e.g., a solid tumor) expresses ILT2. In some embodiments, metastatic or locally advanced cancer (e.g., a solid tumor) expresses ILT2. In some embodiments, the cancer (e.g., a solid tumor) expresses PD-1. In some embodiments, metastatic or locally advanced cancer (e.g., a solid tumor) expresses PD-1. In some embodiments, the cancer (e.g., a solid tumor) expresses PD-L1. In some embodiments, metastatic or locally advanced cancer (e.g., a solid tumor) expresses PD-L1. In some embodiments, the cancer (e.g., a solid tumor) expresses CTLA-4. In some embodiments, metastatic or locally advanced cancer (e.g., a solid tumor) expresses CTLA-4.

[0155] Cancers that can be treated according to the methods described herein include, but are not limited to: solid cancers (e.g., recurrent or refractory solid cancers, and advanced or metastatic solid cancers), carcinomas, sarcomas, melanomas (e.g., stage III or IV melanomas), small cell lung cancer, non-small cell lung cancer, urothelial carcinoma, ovarian cancer, prostate cancer (e.g., metastatic hormone-refractory prostate cancer and advanced metastatic prostate cancer), pancreatic cancer, and breast cancer (e.g., HER2-positive prostate cancer). + Breast cancer (e.g., relapsed / refractory HER2+ breast cancer), head and neck cancer (e.g., relapsed / refractory head and neck squamous cell carcinoma (HNSCC)), glioma, malignant glioma, glioblastoma multiforme, brain metastases, Merkel cell carcinoma, gastric cancer, gastroesophageal junction cancer, renal cell carcinoma, uveal melanoma, colon cancer, cervical cancer, lymphoma (e.g., relapsed or refractory lymphoma), non-Hodgkin lymphoma, Hodgkin lymphoma, leukemia, and multiple myeloma.

[0156] In some implementations, the cancers treated according to the methods described herein are B-cell lymphomas (e.g., B-cell chronic lymphocytic leukemia, B-cell non-Hodgkin lymphoma, cutaneous B-cell lymphoma, diffuse large B-cell lymphoma), basal cell carcinoma, bladder cancer, blastoma, brain metastases, breast cancer, Burkitt lymphoma, carcinomas (e.g., adenocarcinoma (e.g., gastroesophageal junction adenocarcinoma)), cervical cancer, colon cancer, colorectal cancer (colon and rectal cancer), and intrauterine cancer. Membrane cancer, esophageal cancer, Ewing sarcoma, follicular lymphoma, gastric cancer, gastroesophageal junction cancer, gastrointestinal cancer, glioblastoma (e.g., glioblastoma multiforme, such as newly diagnosed or recurrent), glioma, head and neck cancer (e.g., head and neck squamous cell carcinoma), liver metastases, Hodgkin lymphoma and non-Hodgkin lymphoma, renal cell carcinoma (e.g., renal cell carcinoma and nephroblastoma), laryngeal cancer, leukemia (e.g., chronic myeloid leukemia, hairy cell leukemia), liver cancer ( For example, hepatocellular carcinoma and hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), lymphoblastic lymphoma, lymphoma, mantle cell lymphoma, metastatic brain tumors, metastatic cancer, myeloma (e.g., multiple myeloma), neuroblastoma, ocular melanoma, oropharyngeal carcinoma, osteosarcoma, ovarian cancer, pancreatic cancer (e.g., pancreatic ductal adenocarcinoma), prostate cancer (e.g., hormone-refractory (e.g., castration-resistant), metastatic, metastatic hormone-refractory (e.g., castration-resistant, androgen-independent)), renal cell carcinoma (e.g., metastatic), salivary gland cancer, sarcoma (e.g., rhabdomyosarcoma), skin cancer (e.g., melanoma (e.g., metastatic melanoma)), soft tissue sarcoma, solid tumors, squamous cell carcinoma, synovial sarcoma, testicular cancer, thyroid cancer, transitional cell carcinoma (urothelial carcinoma), uveal melanoma (e.g., metastatic), verrucous carcinoma, vulvar cancer, and Warburg's macroglobulinemia.

[0157] In some implementations, the cancer treated according to the methods described herein is a human sarcoma or carcinoma, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovoma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma. Medullary carcinoma, bronchial carcinoma, renal cell carcinoma (e.g., metastatic), hepatocellular carcinoma, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, nephroblastoma, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, glioblastoma multiforme, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineal tumor, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, or retinoblastoma.

[0158] In some implementations, the cancer treated according to the methods described herein is acute lymphoblastic leukemia or acute myeloid leukemia (e.g., myeloblastic, promyelocytic, granulocytic-monocytic, monocytic, and erythroleukemia); chronic leukemia (chronic myeloid (granulocytic) leukemia or chronic lymphocytic leukemia); Hodgkin's disease; non-Hodgkin's disease; acute myeloid leukemia; B-cell lymphoma; T-cell lymphoma; anaplastic large cell lymphoma; intraocular lymphoma; follicular lymphoma; small intestinal lymphoma; or splenic marginal zone lymphoma.

[0159] In some implementations, the cancers treated according to the methods described herein are multiple myeloma, Warren's macroglobulinemia, heavy chain disease, gastrointestinal stromal tumors, head and neck cancers (e.g., hypopharyngeal squamous cell carcinoma, laryngeal squamous cell carcinoma, oropharyngeal cell carcinoma, or laryngeal verrucous carcinoma), endometrial stromal sarcoma, mast cell sarcoma, adult soft tissue sarcoma, uterine sarcoma, Merkel cell carcinoma, urothelial carcinoma, melanoma with brain metastases, uveal melanoma, uveal melanoma with liver metastases, non-small cell lung cancer, rectal cancer, or myelodysplastic syndrome. In some implementations, the cancers treated according to these methods are metastatic.

[0160] In some embodiments, the cancers treated according to the methods described herein are prostate cancer, breast cancer, lung cancer, colorectal cancer, melanoma, bronchial cancer, bladder cancer, brain or central nervous system cancer, peripheral nervous system cancer, uterine or endometrial cancer, oral or pharyngeal cancer, non-Hodgkin lymphoma, thyroid cancer, kidney cancer, biliary tract cancer, small bowel or appendix cancer, salivary gland cancer, thyroid cancer, adrenal cancer, squamous cell carcinoma, mesothelioma, bone cancer, thymoma / thymic carcinoma, glioblastoma, myelodysplastic syndrome, soft tissue sarcoma, diffuse engenerative pontine glioma (DIPG), adenocarcinoma, osteosarcoma, chondrosarcoma, leukemia, or pancreatic cancer. In some embodiments, the cancers treated according to the methods described herein include carcinoma (e.g., adenocarcinoma), lymphoma, blastoma, melanoma, sarcoma, or leukemia.

[0161] In some embodiments, the cancers treated according to the methods described herein are squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, gastrointestinal cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, pancreatic cancer, glioblastoma, glioma, cervical cancer, ovarian cancer, liver cancer (e.g., hepatocellular carcinoma and hepatocellular carcinoma), bladder cancer, breast cancer, inflammatory breast cancer, Merkel cell carcinoma, colon cancer, colorectal cancer, stomach cancer, bladder cancer, endometrial cancer, myeloma (e.g., multiple myeloma), salivary gland cancer, kidney cancer (e.g., renal cell carcinoma and nephroblastoma), basal cell carcinoma, melanoma, prostate cancer, vulvar cancer, thyroid cancer, testicular cancer, esophageal cancer, serous adenocarcinoma, or various types of head and neck cancer. In some embodiments, the cancers treated according to the methods described herein include desmoplastic melanoma, inflammatory breast cancer, thymoma, rectal cancer, anal cancer, or surgically treatable or inoperable brainstem gliomas.

[0162] In some embodiments, the cancer treated according to the methods described herein is metastatic melanoma (e.g., drug-resistant metastatic melanoma), metastatic ovarian cancer, or metastatic renal cell carcinoma. In some embodiments, the cancer treated according to the methods described herein is melanoma resistant to ipilimumab. In some embodiments, the cancer treated according to the methods described herein is melanoma resistant to nivolumab or pembrolizumab. In some embodiments, the cancer treated according to the methods described herein is melanoma resistant to both ipilimumab and nivolumab or pembrolizumab.

[0163] In some implementations, the cancers treated according to the methods described herein are breast cancer (e.g., Herceptin-resistant breast cancer and trastuzumab-DM1 (T-DM1)-resistant breast cancer), prostate cancer, glioblastoma multiforme, colorectal cancer, sarcoma, bladder cancer, cervical cancer, HPV-related cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, oropharyngeal cancer, multiple myeloma, renal cell carcinoma, ovarian cancer, hepatocellular carcinoma, endometrial cancer, pancreatic cancer, lymphoma, and leukemia (e.g., senile leukemia, acute myeloid leukemia (AML), and senile AML).

[0164] In some implementations, the cancers treated according to the methods described herein are metastatic malignant melanoma (e.g., cutaneous or ocular malignant melanoma), renal cell carcinoma (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), breast cancer, colon cancer, lung cancer (e.g., non-small cell lung cancer), bone cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, esophageal cancer, small bowel cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, and vaginal cancer. Stem cell carcinoma, chronic or acute leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia), childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) tumors, primary CNS lymphoma, tumor angiogenesis, spinal tumors, brainstem glioma, glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancer (including asbestos-induced cancer), esophageal cancer, liver cancer, refractory or recurrent malignant tumors, metastatic cancer, and combinations thereof.

[0165] In some implementations, the subject has previously received immunotherapy. In some implementations, the subject has not previously received any immunotherapy. In some implementations, the cancer is advanced or metastatic.

[0166] In some implementations, the subject's age is at least 18 years. In some implementations, the subject is histologically diagnosed with a solid tumor that is currently metastatic or locally advanced. In some implementations, no standard therapy is available or standard therapy has failed. In some implementations, the subject has measurable disease on baseline imaging prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor, as assessed according to the Recognition of Efficacy in Solid Tumors version 1.1 (RECIST 1.1). In some implementations, the subject's life expectancy... > Three months. In some implementations, the subject's Eastern Oncology Cooperative Group performance status score was 0-1 prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor. In some implementations, the subject had adequate organ and bone marrow reserve function, as defined by one or more of the following, prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor: absolute neutrophil count. > 1.5×10 9 / L; platelet count > 100×10 9 / L; Hemoglobin >8.0 g / dL, and no blood transfusion within 2 weeks prior to hemoglobin measurement; serum albumin > 3.0 g / dL; Total bilirubin < 1.5 × Upper Limit of Normal (ULN); Aspartate Aminotransferase < 2.5×ULN; alanine aminotransferase < 2.5×ULN; alkaline phosphatase < 2.5×ULN, or for subjects with liver metastases < 5.0 × ULN; Creatinine clearance > 40 mL / min, as assessed by the Cockcroft-Gault method (see Cockcroft DW, Gault MH. Nephron. 1976; 16(1):31-41); and / or international normalized ratio or prothrombin time. < 1.5×ULN and activation time of partial thromboplastin < 1.5 × ULN, unless the patient is receiving anticoagulation therapy. In some implementations, the subject is not pregnant and / or not breastfeeding.

[0167] In some embodiments, the subject has not received cytotoxic therapy or other monoclonal antibody therapy within 3 weeks prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor. In some embodiments, the subject has not received central nervous system (CNS) radiation therapy within 2 weeks prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor, or has not received non-CNS radiation therapy within 1 week prior to administration. In some embodiments, the subject has not received small molecule targeted therapy or tyrosine kinase inhibitor therapy within 2 weeks prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor or within 5 half-lives of the therapy.

[0168] In some implementation schemes, the subject does not have any persistent toxicity from prior cancer therapy (Common Terminology Standard for Adverse Events [CTCAE]). >Grade 1). In some implementations, the subject does not have grade >2 sensory neuropathy or alopecia. In some implementations, the subject does not have a history of interstitial lung disease or active interstitial lung disease. In some implementations, the subject does not have a history of allergic reactions or uncontrolled asthma. In some implementations, the subject does not have a condition requiring systemic treatment with corticosteroids (daily prednisone equivalent >10 mg) within 2 weeks prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor, or with other immunosuppressive drugs within 30 days prior to administration. In some implementations, the subject does not have active CNS metastases unless the CNS metastases have been treated and the subject is radiographically and clinically stable. In some implementations, the subject does not have an active autoimmune disease or a history of autoimmune disease requiring systemic treatment within 2 years prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor, unless the treatment includes hormone replacement therapy or topical treatment. In some implementations, the subject has not received an allogeneic tissue or solid organ transplant, except for corneal transplantation.

[0169] In some implementations, the subject does not have an active infection requiring treatment within 2 weeks prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor. In some implementations, the subject is not HIV-positive unless the subject's CD4 count is >200 cells / mm². 3 Or the viral load may be undetectable. In some implementations, the subject does not have active hepatitis B (HBV) or hepatitis C (HPC) unless their hepatitis B surface antigen and HBV DNA or HCV RNA are negative, respectively. In some implementations, the subject does not have clinically significant cardiovascular disease, as defined by one or more of the following: cerebrovascular accident or stroke or myocardial infarction within 6 months prior to administration of the primary antibody and / or human PD-1 inhibitor and / or human CTLA-4 inhibitor; unstable angina; congestive heart failure (New York Heart Association classification). > Grade II); and / or uncontrolled arrhythmias requiring medication. In some implementations, the subject's QTc interval is not greater than 480 milliseconds, unless the QTc prolongation is due to right bundle branch block. In some implementations, the subject does not have uncontrolled hypertension (>140 / 90 mmHg) or hypertension requiring more than three antihypertensive medications.

[0170] In some embodiments, the primary antibody is administered as a single therapy. In some embodiments, the primary antibody is administered on the same day as a human PD-1 inhibitor. In some embodiments, the primary antibody is administered on the same day as a human CTLA-4 inhibitor. In some embodiments, the primary antibody is administered on the same day as both a human PD-1 inhibitor and a human CTLA-4 inhibitor. In some embodiments, each administration of the primary antibody and the human PD-1 inhibitor is on the same day. In some embodiments, each administration of the human CTLA-4 inhibitor and the primary antibody is on the same day. In some embodiments, the human PD-1 inhibitor is administered 30 minutes or 1 hour after the primary antibody administration. In some embodiments, the human CTLA-4 inhibitor is administered 30 minutes or 1 hour after the primary antibody administration. In some embodiments, the human CTLA-4 inhibitor is administered 30 minutes or 1 hour after the human PD-1 inhibitor administration.

[0171] In some embodiments, the primary antibody is administered at a dose of 1 mg to 1000 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg. In some embodiments, the primary antibody is administered at a dose of 1 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg. In some embodiments, the primary antibody is administered at a dose of 3 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg. In some embodiments, the primary antibody is administered at a dose of 10 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg. In some embodiments, the primary antibody is administered at a dose of 30 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg. In some embodiments, the primary antibody is administered at a dose of 1000 mg, and the human PD-1 inhibitor is administered at a dose of 450 mg.

[0172] In some embodiments, the primary antibody is administered at a dose of 1 mg to 1000 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 1 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 3 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 10 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 30 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 1000 mg, and the human CTLA-4 inhibitor is administered at a dose of 150 mg.

[0173] In some embodiments, the primary antibody is administered at a dose of 1 mg to 1000 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 1 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 3 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 10 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 30 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at a dose of 100 mg, the human PD-1 inhibitor at a dose of 450 mg, and the human CTLA-4 inhibitor at a dose of 150 mg. In some embodiments, the primary antibody is administered at 300 mg, the human PD-1 inhibitor at 450 mg, and the human CTLA-4 inhibitor at 150 mg. In some embodiments, the primary antibody is administered at 1000 mg, the human PD-1 inhibitor at 450 mg, and the human CTLA-4 inhibitor at 150 mg.

[0174] The primary antibody, human PD-1 inhibitor, and human CTLA-4 inhibitor may be administered simultaneously or sequentially via the same or different routes of administration (e.g., as described herein). In some embodiments, the primary antibody, human PD-1 inhibitor, and human CTLA-4 inhibitor are administered simultaneously via intravenous injection (e.g., intravenous infusion). In some embodiments, the primary antibody, human PD-1 inhibitor, and human CTLA-4 inhibitor are administered sequentially via intravenous injection (e.g., intravenous infusion). In some embodiments, the primary antibody is administered intravenously at doses of 1 mg to 1000 mg, and the human PD-1 inhibitor is administered intravenously at doses of 450 mg, every three weeks. In some embodiments, the primary antibody is administered intravenously at doses of 1 mg to 1000 mg, and the human CTLA-4 inhibitor is administered intravenously at doses of 150 mg, every three weeks. In some embodiments, the primary antibody is administered intravenously at doses of 1 mg to 1000 mg every three weeks, the human PD-1 inhibitor is administered intravenously at doses of 450 mg every three weeks, and the human CTLA-4 inhibitor is administered at doses of 150 mg every six weeks.

[0175] In some embodiments, the method includes administering a first antibody to the subject, such as an antibody comprising the heavy and light chains of the amino acid sequences SEQ ID NO: 9 and 10, respectively; a human PD-1 inhibitor (e.g., bitetimab); and a human CTLA-4 inhibitor (e.g., botelimab). In some embodiments, bitetimab is administered by intravenous infusion at a dose of 450 mg every 3 weeks. In some embodiments, botelimab is administered by intravenous infusion at a dose of 150 mg every 6 weeks.

[0176] In some embodiments, bartelimab is administered via intravenous infusion at a dose of 450 mg every 3 weeks, and botulinumab is administered via intravenous infusion at a dose of 150 mg every 6 weeks. The primary antibody is administered at doses of approximately 1 mg to approximately 2000 mg, approximately 1 mg to approximately 1000 mg, approximately 3 mg to approximately 300 mg, approximately 10 mg to approximately 100 mg, approximately 1 mg to approximately 600 mg, approximately 1 mg to approximately 300 mg, approximately 1 mg to approximately 100 mg, approximately 1 mg to approximately 30 mg, approximately 1 mg to approximately 10 mg, approximately 1 mg to approximately 3 mg, and approximately 10 mg to approximately 2000 mg. Administered in doses of 00 mg, about 10 mg to about 1000 mg, about 10 mg to about 600 mg, about 10 mg to about 300 mg, about 10 mg to about 30 mg, about 30 mg to about 1000 mg, about 30 mg to about 600 mg, about 30 mg to about 300 mg, about 30 mg to about 100 mg, about 100 mg to about 1000 mg, about 100 mg to about 600 mg, about 100 mg to about 300 mg, about 300 mg to about 2000 mg, about 300 mg to about 1000 mg, or about 300 mg to about 600 mg. In some implementations, bartelimab is administered via intravenous infusion at a dose of 450 mg every 3 weeks, botulinumab is administered via intravenous infusion at a dose of 150 mg every 6 weeks, and the primary antibody is administered at doses of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1250 mg, about 1500 mg, about 1750 mg, or about 2000 mg.

[0177] In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered via intravenous infusion over approximately 30 minutes.

[0178] In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, or every 6 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 1 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 3 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 10 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 30 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 100 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 300 mg every 3 weeks. In some embodiments, bitemimab is administered intravenously at a dose of 450 mg every 3 weeks, botelimab is administered intravenously at a dose of 150 mg every 6 weeks, and the primary antibody is administered intravenously at a dose of 1000 mg every 3 weeks.

[0179] In one aspect, this article provides an antibody that specifically binds to human ILT2 for use in the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human PD-1 inhibitor for use in the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human CTLA-4 inhibitor for use in the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2, a human PD-1 inhibitor, and a human CTLA-4 inhibitor for use in the treatment of cancer, wherein the treatment is performed according to the methods described herein.

[0180] In one aspect, this article provides an antibody that specifically binds to human ILT2 for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human PD-1 inhibitor for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human CTLA-4 inhibitor for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2, a human PD-1 inhibitor, and a human CTLA-4 inhibitor for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the methods described herein.

[0181] In one aspect, this article provides an antibody that specifically binds to human ILT2 for the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human PD-1 inhibitor for the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2 and a human CTLA-4 inhibitor for the treatment of cancer, wherein the treatment is performed according to the methods described herein. In one aspect, this article provides an antibody that specifically binds to human ILT2, a human PD-1 inhibitor, and a human CTLA-4 inhibitor for the treatment of cancer, wherein the treatment is performed according to the methods described herein.

[0182] Example The embodiments in this section are provided by way of illustration rather than by way of limitation.

[0183] Example 1: A study in patients with advanced solid tumors of AAGEN1571 as monotherapy and in combination with PD-1 inhibitors and / or other drugs. Phase 1 study of combination therapy with orthopedic adenocarcinoma (AGEN1181) AGEN1571 is a fully human monoclonal IgG4 antibody that antagonizes ILT2. This example describes a multicenter, phase 1 trial evaluating the safety, tolerability, pharmacokinetics, and pharmacodynamics of AGEN1571 as monotherapy and in combination with bartelimab and / or botulinumab for the treatment of patients with advanced or metastatic solid tumors to determine the recommended phase 2 dose (RP2D).

[0184] Overall research design This is an open-label, phase 1, two-part trial to determine the RP2D of AAGEN1571 as monotherapy and in combination with baritelimab and / or botulinumab in patients diagnosed with advanced solid tumors, and to evaluate its safety, tolerability, PK, and pharmacodynamic characteristics.

[0185] The study will recruit up to 98 evaluable patients. The actual number of patients enrolled will depend on safety, tolerability, and duration of treatment.

[0186] Part 1 is the dose escalation phase, used to determine the RP2D for either AREN1571 monotherapy or AREN1571 in combination with either bartelimab or botelimab (two-drug combination) or both bartelimab and botelimab (three-drug combination).

[0187] Part 2 of the trial was a dose expansion phase for a specific disease indication.

[0188] Goals and End Points Table 4. Goals and Endpoints

[0189] Research on treatment Part 1 - Dosage Escalation Phase Part 1 of the study will consist of four study treatment groups. Study treatment group 1 (AGEN1571 monotherapy) will use an accelerated titration design for AGEN1571 monotherapy to assign patients to cohorts 1 through 3, followed by cohorts 4 through 7 using a conventional 3+3 design.

[0190] After determining the active dose of AREN1571 monotherapy (Study Treatment Group 1), a 3+3 dose escalation design will be used to evaluate combination therapy of AREN1571 (Q3W dosing) with baritelimab and / or botelimab as described below. Patients will be assigned to Study Treatment Groups 2 through 4 using a standard 3+3 dose escalation design of AREN1571 in combination with baritelimab and / or botelimab.

[0191] If the treatment criteria are not met within the treatment window, the dose will be skipped, and the patient must wait until the next scheduled dose to resume treatment. The study treatment group is as follows: Table 5. Study Treatments

[0192] Patients diagnosed with advanced solid tumors will be assigned to cohorts using an interactive response technology system. Alternative dosing regimens for AREN1571 may also be explored based on pharmacodynamic and safety results. The maximum recommended starting dose for AREN1571, administered intravenously (IV) every 3 weeks, is 1 mg (approximately 0.014 mg / kg in a 70 kg patient), derived based on the 80% receptor occupancy (RO) method outlined by FDA scientists.

[0193] For Part 1 of the study, once the active dose of AGEN1571 as monotherapy or in combination with bartelimab and / or botulinumab is determined, an additional 15 evaluable patients may be added to the dose escalation cohort in any of the SMC-approved treatment groups (study treatment groups 1 to 4) to further evaluate safety, pharmacodynamics, and PK parameters.

[0194] With SMC approval, dose reduction is permitted between different cohorts across all study treatment groups. Intra-patient dose reduction or modification is not permitted.

[0195] Part 2 - Dosage Expansion Phase for Specific Disease Indications Based on data from Part 1 (dose escalation phase) and at the sponsor's discretion, Part 2 of this study will be an open-ended expansion cohort by tumor type. Part 2 will enroll at least 10 and up to approximately 30 patients by tumor type at doses of SMC-approved monotherapy or any combination therapy. The aim is to further confirm the safety of the dose, meet the need for additional PK and / or additional pharmacodynamic data, further evaluate any significant efficacy observed during the dose escalation phase that may include responses related to pharmacodynamic signals and manifested as long-term disease stability, and assess the clinical activity observed in patients at previously SMC-approved doses.

[0196] All inclusion / exclusion criteria will remain unchanged in the expanded cohort. The expanded cohort follows the same safety criteria outlined for the dose escalation cohort. Specifically, intra-patient dose modification is not permitted.

[0197] Number of patients For Part 1 (dose escalation phase), up to 98 DLT-evaluable patients will be enrolled. For Part 2 (dose expansion phase), each dose expansion cohort will enroll at least 10 and up to approximately 30 patients based on clinical considerations.

[0198] Inclusion criteria To participate in this study, patients should meet all of the following inclusion criteria: 1. Prior to any research-specific procedure, participants voluntarily consent to participate by providing signed, dated, and written informed consent (participation in genetic testing is optional).

[0199] 2. Age > 18 years old.

[0200] 3. Histologically confirmed to have a solid tumor that is currently metastatic or locally advanced, for which there is no standard treatment available or the standard treatment has failed.

[0201] 4. Based on RECIST 1.1, there is a measurable disease on baseline imaging.

[0202] 5. Life expectancy of at least 3 months.

[0203] 6. The ECOG performance status is 0 or 1.

[0204] 7. Sufficient organ and bone marrow reserve function, as indicated by the following laboratory values: a. Adequate hematological function, defined as an absolute neutrophil count. > 1.5×10 9 / L, platelet count > 100×10 9 / L, hemoglobin > 8 g / dL, and no recent blood transfusion (recent blood transfusion is defined as a blood transfusion that occurred within 2 weeks prior to the hemoglobin measurement).

[0205] b. Adequate liver function, defined as serum albumin > 3.0 g / dL, total bilirubin level < 1.5 × Upper Limit of Normal (IULN), Aspartate Aminotransferase < 2.5×IULN, alanine aminotransferase < 2.5 × IULN, alkaline phosphatase < 2.5 × IULN, or for patients with liver metastases < 5×ULN.

[0206] c. Adequate renal function, defined as creatinine clearance calculated as assessed by the Cockcroft-Gault method. > 40 mL / min.

[0207] d. Adequate coagulation function, defined as international normalized ratio or prothrombin time. < 1.5×IULN and activated partial thromboplastin time< 1.5×IULN (unless the patient is receiving anticoagulation therapy).

[0208] 8. Patients with a history of malignancy, if their treatment was prior to the first dose of study treatment. > If the two-year period has been completed and the patient has no evidence of disease, then the patient is eligible. Patients who have been continuously receiving stable doses of anti-tumor hormone therapy during these two years may also be eligible with the approval of a medical monitor.

[0209] Patients with a history of early basal / squamous cell skin cancer, or non-invasive or in situ carcinoma, and who have received radical treatment at any time, are also eligible.

[0210] 9. The patient must provide a full and adequate formalin-fixed paraffin-embedded tumor tissue sample (biopsy) taken from a previously untreated site, obtained after the last dose of the previous anticancer treatment and before the first dose of the study treatment, and agree to mandatory in-treatment biopsy where clinically feasible.

[0211] If a potential study participant is unable to provide a tumor tissue sample as specified above, they may still be enrolled after discussion and approval by the medical monitor.

[0212] 10. Female patients of childbearing potential must have a negative urine or serum pregnancy test at screening (within 72 hours of the first dose of the study drug) and repeat the urine or serum pregnancy test on day 1 of each cycle and at the end-of-treatment visit. If the urine pregnancy test is positive, the result must be confirmed by a serum pregnancy test. Infertility is defined as: a. Age > 50 years old and has not menstruated for more than one year.

[0213] b. Amenorrhea > Two years prior, without hysterectomy or bilateral oophorectomy, and with follicle-stimulating hormone (FSH) levels in the postmenopausal range at the pre-study (screening) evaluation.

[0214] c. Has undergone hysterectomy, bilateral oophorectomy, or tubal ligation.

[0215] Female patients diagnosed with a known tumor that secretes human chorionic gonadotropin (hCG) must be confirmed as not pregnant based on clinical evidence and the investigator's judgment.

[0216] Female patients of childbearing potential must agree to use highly effective contraception from the start of the screening visit until 90 days after the last dose of study treatment. Definitions of highly effective contraception are given in Appendix B, Contraception Guidelines, or as provided in national or local guidelines.

[0217] Note: Abstinence is acceptable if it is the patient's preferred method of contraception.

[0218] 11. Male patients with female partners of childbearing potential must agree to use highly effective contraception throughout the study period, from the start of the screening visit until 90 days after the last dose of study treatment. Men whose partners become pregnant must agree to use condoms; no additional contraception is required for pregnant partners.

[0219] Note: Abstinence is acceptable if it is the patient's preferred method of contraception.

[0220] 12. Willing and able to meet the requirements of the agreement.

[0221] Exclusion criteria Patients will not be eligible for enrollment in this study if they meet any of the following criteria: 1. Currently participating in and receiving study treatment, or having participated in and received study treatment or used study equipment within 3 weeks prior to the first administration of the current study drug.

[0222] 2. Prior to the first administration of the study drug, outside of an acceptable washout period, the patient had received previous systemic cytotoxic chemotherapy, biotherapy, radiotherapy, or major surgery.

[0223] From the start of previous treatment, the following washout window is acceptable; patients with a washout period shorter than the following should be excluded: a. Cytotoxic drugs or monoclonal antibodies > Three weeks is acceptable (i.e., <3 weeks should be excluded).

[0224] b. For palliative radiotherapy for non-central nervous system (CNS) diseases, a 1-week washout period is permitted.

[0225] c. Small molecule targeted therapy and tyrosine kinase inhibitors > 14 days or 5 half-lives is acceptable (i.e., <14 days or <5 half-lives should be excluded).

[0226] d. It is acceptable to have received a SARS-CoV-2 vaccine >7 days prior to administration of the study drug. For vaccines requiring more than one dose, the entire series should be completed before Day 1 of Cycle 1, where feasible and provided that delaying the start of the study treatment would not put the study patients at risk.

[0227] e. Has received prior CNS radiation therapy within 2 weeks prior to the first treatment.

[0228] 3. There is persistent toxicity related to previous treatment, with a severity level >1 according to NCI CTCAE version 5.0.

[0229] 4. < Grade 2 sensory neuropathy or alopecia are acceptable. Stable compensatory endocrine disorders (hypothyroidism, adrenal insufficiency, hypopituitarism) as sequelae of immune-mediated adverse events >1 are acceptable. Known serious adverse events (such as those with fully human monoclonal antibodies or any excipients used in investigational drugs) are also acceptable. > Grade 3 hypersensitivity reaction, or severe reaction to immuno-oncology drugs, such as colitis or pneumonia requiring steroid treatment; or a history of interstitial lung disease or active interstitial lung disease, any history of allergic reactions, or uncontrolled asthma.

[0230] 5. Patients with a condition requiring systemic treatment with corticosteroids (daily prednisone equivalent >10 mg) within 14 days prior to the first dose of study treatment or with any other immunosuppressive medication within 30 days prior to the first dose. In the absence of active autoimmune disease, inhaled or topical steroids, as well as adrenal replacement doses of steroids (daily prednisone equivalent), are permitted. < 10mg).

[0231] 6. Active CNS metastasis.

[0232] Note: Patients are eligible if CNS metastases have been treated and the patient is stable both radiographically and clinically. Patients must have discontinued corticosteroids for at least 2 weeks prior to the first dose of study treatment, or have a stable or tapered daily dose of prednisone or its equivalent. < 10mg.

[0233] 7. A history of active autoimmune disease or autoimmune disorders requiring systemic treatment (i.e., use of disease modifiers, corticosteroids, or immunosuppressive drugs) within 2 years prior to the start of treatment with the study drug.

[0234] Note: Patients with autoimmune diseases who require hormone replacement therapy or local treatment are eligible.

[0235] 8. Has received allogeneic tissue / solid organ transplants, excluding corneal transplants.

[0236] 9. Active infection or active interstitial lung disease that requires treatment within 2 weeks prior to Day 1 of Cycle 1 (C1D1).

[0237] 10. HIV test positive.

[0238] a. CD4 > 200 cells / mm 3 The patient met the criteria.

[0239] b. Patients with undetectable HIV viral load are eligible.

[0240] 11. Active hepatitis B (HBV) or active hepatitis C (HCV).

[0241] a. HBV-infected patients who are negative for both hepatitis B surface antigen and HBV DNA are eligible.

[0242] b. HCV-infected patients who are negative for HCV RNA are eligible.

[0243] 12. Clinically significant (i.e., active) cardiovascular disease: cerebrovascular accident / stroke or myocardial infarction, unstable angina, or congestive heart failure (New York Heart Association classification) within the 6 months prior to enrollment. > Grade II), or serious uncontrolled arrhythmias requiring medication.

[0244] 13. History or current evidence of any medical condition (including mental disorders or substance abuse disorders that may interfere with the trial requirements), any treatment that may interfere with the trial, any active infection that requires treatment within 2 weeks prior to C1D1, or any laboratory abnormalities that may confound the trial results, interfere with the patient's full participation in the trial, or that the attending investigator deems participation in the trial not to be in the patient's best interest.

[0245] 14. Lacking legal capacity or having limited legal capacity.

[0246] 15. Pregnant or breastfeeding.

[0247] 16. Screening is performed if the QTc interval is >480 milliseconds, unless the QTc prolongation is due to right bundle branch block.

[0248] 17. Uncontrolled hypertension, and hypertension requiring more than 3 antihypertensive medications (>140 / 90 mmHg).

[0249] Product, dosage and administration method AGEN1571: Q3W (i.e., on days 1 and 22 of each 6-week cycle) will be administered via IV infusion over 30 (±5) minutes. Batelimab: Q3W (i.e., on days 1 and 22 of each 6-week cycle) will be administered via IV infusion over 30 (±5) minutes following administration of AAGEN1571. * Bortimab: Q6W (i.e., day 1 of each 6-week cycle) will be administered via IV infusion over 30 (±5) minutes following administration of either AGEN1571 or bartimab. * *All deviations from the infusion duration window will be subject to medical review, and the impact of extending or shortening the infusion time on clinical and research significance will be assessed. If there is no impact on patient safety or any key research endpoints (e.g., PK data interpretation), these deviations will not be considered protocol deviations at all, but rather normal variations in clinical practice.

[0250] Study group 1: AAGEN 1571 monotherapy AGEN1571 will be evaluated as monotherapy in 1 to 6 patients in three dose cohorts (1 mg, 3 mg, and 10 mg) using a fixed-dose accelerated titration design. After the accelerated titration phase, AGEN1571 will be evaluated as monotherapy in four dose cohorts (30 mg, 100 mg, 300 mg, and 1000 mg) using a 3+3 dose escalation design, with 3 to 6 patients per cohort, administered Q3W (days 1 and 22 of each 6-week cycle).

[0251] Table 6. Study Treatment Group 1: AAGEN 1571 Monotherapy

[0252] Abbreviation: Q3W = once every 3 weeks. a Cohorts 1 through 3 will follow an accelerated titration design for dose escalation. Cohorts 4 through 7 will follow a 3+3 design for dose escalation. b Based on all safety data, and with the consent of the sponsor and investigators, the planned dose level of AGEN1571 monotherapy may be reduced, or the intermediate dose level may be increased.

[0253] Note: Alternative dosing regimens for AGEN1571 may be explored based on safety, preliminary efficacy, pharmacokinetic, and pharmacodynamic data.

[0254] Note: After the active dose is determined in study treatment group 1, up to 15 additional evaluable patients may be enrolled to this active dose cohort for treatment from a safety / PK / pharmacodynamic perspective.

[0255] Study group 2: AAGEN1571 + Batilimab The combination of AGEN1571 and bartelimab will be evaluated using a 3+3 dose escalation design in two dose cohorts.

[0256] Table 7. Study group 2: AAGEN1571 + Batelimab

[0257] Abbreviations: DLT = Dose-limiting toxicity; PK = Pharmacokinetics; Q3W = Once every 3 weeks; SMC = Safety Monitoring Committee.

[0258] Note: For combination therapy, AGEN1571 should be administered before bartelimab, with a 30-minute interval between the two infusions.

[0259] Note: Alternative dosing regimens for AGEN1571 and bartelimab may be explored based on safety, preliminary efficacy, pharmacokinetic, and pharmacodynamic data.

[0260] Note: Once the active dose is determined, up to 15 additional evaluable patients can be treated with the active dose of AAGEN1571 + Batelimab, taking into account safety, pharmacodynamics, and other factors.

[0261] Study group 3: AAGEN1571 + Bortelimab The combination of AGEN1571 and botelimab will be evaluated using a 3+3 dose escalation design in two dose cohorts.

[0262] Table 8. Study group 3: AAGEN1571 + Bortelimab

[0263] Abbreviations: DLT = Dose-limiting toxicity; PK = Pharmacokinetics; Q3W = Every 3 weeks; Q6W = Every 6 weeks; SMC = Safety Monitoring Committee. *Note: If the SMC deems it appropriate based on data from other studies, a lower dose (e.g., 50 mg, 75 mg, or 100 mg) of botulinumab may be used. The use of a lower dose will apply to all relevant cohorts.

[0264] Note: For combination therapy, AGEN1571 should be administered before botelimab, with a 30-minute interval between the two infusions.

[0265] Note: Alternative dosing regimens for AGEN1571 and botulinumab may be explored based on safety, preliminary efficacy, pharmacokinetic, and pharmacodynamic data.

[0266] Note: Once the active dose is determined, up to 15 additional evaluable patients can be treated with the active dose of AAGEN1571 + Bortelimab, taking into account safety, pharmacodynamics, and other factors.

[0267] Study group 4: AGEN1571 + Batilimab + Bortelimab AGEN1571 in combination with bartelimab and botulinumab will be evaluated in two dose cohorts using a 3+3 dose escalation design.

[0268] Table 9. Study Treatment Group 4: AAGEN1571 + Batilimab + Bortelimab

[0269] Abbreviations: DLT = Dose-limiting toxicity; PK = Pharmacokinetics; Q3W = Every 3 weeks; Q6W = Every 6 weeks; SMC = Safety Monitoring Committee. *Note: If the SMC deems it appropriate based on data from other studies, a lower dose (e.g., 50 mg, 75 mg, or 100 mg) of botulinumab may be used. The use of a lower dose will apply to all relevant cohorts.

[0270] Note: For combination therapy, AGEN1571 should be administered before baritelimab, with a 30-minute interval between the two infusions. Then, baritelimab should be infused 30 minutes after the baritelimab infusion is completed.

[0271] Note: Alternative dosing regimens may be explored based on safety, preliminary efficacy, pharmacokinetic, and pharmacodynamic data.

[0272] Note: Once the active dose is determined, up to 15 additional evaluable patients can be treated using the active dose of AGEN1571 + Batelimab + Bortelimab, taking into account safety, pharmacodynamics, and pharmacodynamics.

[0273] Treatment duration Treatment with AREN1571 monotherapy or in combination with bartelimab and / or boretlimab will continue until confirmed disease progression, unacceptable toxicity, or up to 2 years, whichever occurs first. Patients will be followed for safety at 30 (±7) days and 90 (±7) days after treatment cessation, and will be followed up every 2 months for at least 12 months from the last administration.

[0274] Evaluation criteria Safety: Safety will be assessed through evaluation of adverse events (AEs) according to NCI CTCAE version 5.0 classification, vital signs (temperature, pulse, respiratory rate, and blood pressure), physical examination, 12-lead ECG, ECOG performance status, and clinical laboratory assessments. DLT will be assessed during the dose escalation phase. A safety management mechanism (SMC) will be established to assess safety, determine dose escalation, and define RP2D.

[0275] Pharmacokinetics: Serum PK parameters for AREN1571 may include (but are not limited to) the maximum observed drug concentration at steady state, the minimum observed drug concentration at steady state, the area under the concentration-time curve (AUC) over the steady-state time span t1 to t2, the AUC from time zero to time t, the AUC from time zero to infinity, time to peak concentration, elimination rate constant, terminal elimination half-life, systemic drug clearance, and volume of distribution. PK will be analyzed using both non-compartmental analysis and compartmental models (e.g., population pharmacokinetics [PopPK]). Additional PK exposure parameters for serum bartelimab and botulinumab will be assessed using PopPK.

[0276] Immunogenicity: Serum ADA will be evaluated to determine its potential impact on PK exposure parameters and biological activity of ARDN1571, bartelimab, and botulinumab. Antibody neutralization will be assessed in ADA-positive samples.

[0277] Pharmacodynamics: Plasma, serum, whole blood samples, and tumor biopsy tissues will be collected for exploratory biomarker analysis.

[0278] Efficacy: Response will be assessed according to RECIST 1.1. Imaging studies will be performed every 9 weeks (±7 days) for the first 12 months after treatment initiation, and then every 12 weeks (±7 days) thereafter.

[0279] Statistical methods The total sample size for Part 1 is expected to be a maximum of 98 evaluable, treated patients. Due to the 3+3 design of the study, the sample size will be determined by the number of dose levels evaluated. The sample size for Part 2 will depend on the number of extended cohorts recommended by SMC (10 to 30 patients per cohort).

[0280] Efficacy endpoints will be based on RECIST v1.1. The following efficacy endpoints will be analyzed and summarized to evaluate the antitumor activity of all combinations: ORR, DOR, DCR, PFS, and OS. PFS, DOR, and OS will be estimated using the Kaplan-Meier method. A waterfall plot of maximum tumor shrinkage for each patient will be presented.

[0281] Safety will be determined by spontaneous reporting of adverse events (AEs) and laboratory test values ​​(hematology, clinical chemistry, coagulation function, and urinalysis). Vital signs, physical examination, and electrocardiogram results will also be used to determine safety characteristics. The severity of AEs will be classified according to the NCI-CTCAE version 5.0. The incidence of DLT events and TEAEs will be reported as the percentage of patients who experienced a TEAE, categorized by organ system and preferred terminology. Descriptive statistics will generally be used to summarize study results; that is, statistics for continuous variables will include the mean, standard deviation, median, minimum, and maximum, and for pharmacokinetic summaries, the coefficient of variation and geometric mean will also be included. Qualitative variables will be summarized by counts and percentages. Unless otherwise specified, proportions will be calculated based on the sample size of the focus analysis set.

[0282] Interim analyses will be conducted as appropriate to assess preliminary safety and activity evidence. These will be exploratory in nature, for descriptive purposes only, and will not involve formal statistical testing.

[0283] Example 2: AAGEN1571 combined with the PD-1 antagonist bitetimab enhances T cell activation, NK cell activation, and NKT activation. Cell activation The combination potential of AGEN1571 with the anti-PD-1 antagonist (batielimab; AGEN2034) was evaluated in an immune cell activation assay, where allogeneic recognition of tumor cells promoted the activity of T cells, NK cells, and NKT cells. Cells from the endogenous HLA-G-expressing human choriocarcinoma JEG3 cell line (Avril et al., J Immunol (1999) 162 (10): 5902-5909) were used as target cells. Peripheral blood mononuclear cells (PBMCs) from healthy donors were used as effector cells. Cryopreserved PBMCs from 10 healthy donors were stimulated overnight with IL-2 and IL-15 in the presence of AGEN1571, AGEN2034, or allotype control antibodies. The next day, JEG-3 cells expressing HLA-G and HLA-A2 were... HLA-A2 Cells were added to stimulated PBMCs (tumor cell to PBMC ratio 1:40) and co-cultured for 72 hours. T cell (CD3+) levels were detected by flow cytometry. + CD4 + and CD3 + CD8 + NK cells (CD3) - CD56 + ) and NKT cells (CD3) + CD56 +Increased CD25 expression was used to assess immune cell activation. Cytokine secretion in the culture supernatant was analyzed by evaluating cytokine levels detected by a microsphere-based immunoassay (Luminex).

[0284] AGEN1571-mediated ILT2 blockade enhances CD8 + T cells, NK (CD3) - CD56 + ) cells and NKT (CD3) + CD56 + The expression of CD25 on cells was reduced, and the combination with the PD-1 inhibitor (batielimab; AGEN2034) further enhanced the activation of NK cells, NKT cells, and cytotoxic CD8+ T cells. Figures 1A to 1C As demonstrated, in 5 out of 6 donors who responded to ARTN1571 treatment, an additive effect of co-blockade of ILT2 (AGEN1571) and PD-1 (batielimab; ARTN2034) was observed, which was assessed by enhanced induction of CD25 expression on NKT cells. Furthermore, as... Figure 1D As shown, the combination of AGEN1571 and AGEN2034 enhanced the secretion of TNFα, IFNγ, and IL-1β cytokines. These data demonstrate the potential therapeutic efficacy of combining AGEN1571 with the PD-1 inhibitor (AGEN2034) in the cancer treatment setting.

[0285] Example 3: AAGEN1571 enhances AAGEN1181-induced FcγR NFAT signal transduction The ability of AGEN1571 to enhance FcγR signaling induced by an Fc-enhanced IgG1κ human monoclonal antibody (AGEN1181) selectively binding to human anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) was tested. Jurkat T cell lines engineered to express (i) human ILT2, (ii) FcγRIIIA receptor (CD16), and (iii) a luciferase reporter gene induced by activated T cell nuclear factor (NFAT) response elements (NFAT) were co-cultured with Raji B cell lines endogenously expressing ILT2 ligands and engineered to express CTLA-4 (RajiCTLA-4).

[0286] In the presence of 0.01 μg / mL to 10 μg / mL of AAGEN 1181 and 10 -4Jurkat ILT2 CD16 NFAT luciferase cells were co-cultured with Raji CTLA-4 cells at concentrations ranging from 1 μg / mL to 5 μg / mL of AGEN1571 or the corresponding IgG4 isotype control antibody. The opsonization effect of AGEN1181 on Raji cells expressing CTLA-4 induced CD16 signaling, which was subsequently inhibited by ILT2 ligand binding. Without being bound by any particular theory, it is assumed that the blockade of ILT2 ligand interaction by AGEN1571 relieved the ILT2-mediated inhibition of AGEN1181-induced CD16 signaling, thereby enhancing luciferase expression. After overnight incubation, luciferase activity was measured as the endpoint for evaluating FcγR signaling.

[0287] like Figure 2A and Figure 2B As shown, ADEN1571 enhanced ADEN1181-induced FcγR stimulation in a dose-dependent manner. Even at the highest test concentration of 10 μg / mL, the allotype control antibody did not increase reporter gene expression. In the absence of ADEN1571, ADEN1181-induced FcγR signaling (luciferase) was negligible, indicating that ILT2 is a potent antagonist of CD16-NFAT signaling. No Fc-enhanced allotype antibody activation of the reporter gene was observed in the presence of ADEN1571. These data demonstrate the potential therapeutic utility of combining ADEN1571 and ADEN1181 in a cancer treatment setting.

[0288] * * * The scope of this invention is not limited to the specific embodiments described herein. In fact, various modifications to the invention will become apparent to those skilled in the art from the foregoing description and drawings, in addition to those described herein. Such modifications are intended to fall within the scope of the appended claims.

[0289] All references cited in this article (e.g., publications, patents, or patent applications) are incorporated herein by reference in their entirety, and for all purposes, to the extent that each individual reference (e.g., publications, patents, or patent applications) is specifically and individually indicated to be incorporated herein by reference in its entirety for all purposes.

[0290] Other implementations are within the scope of the following claims.

Claims

1. A method of treating cancer in a subject in need, the method comprising administering to the subject a first antibody that specifically binds to human ILT2, the first antibody being in a dose of about 1 mg to about 2000 mg, wherein the first antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO:

8.

2. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and VL, wherein the VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and (b) Human PD-1 inhibitors.

3. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and VL, wherein the VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and (b) Human CTLA-4 inhibitors.

4. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and VL, wherein the VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and (b) Human PD-L1 inhibitors.

5. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: VH, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and VL, wherein the VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; and (b) Human PD-L2 inhibitors.

6. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: a heavy chain variable region (VH) comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO: 7; and a light chain variable region (VL) comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO: 8; (b) Human PD-1 inhibitors; and (c) Human CTLA-4 inhibitors.

7. The method according to any one of the preceding claims, wherein the first antibody is administered at a dose of about 1 mg to about 1000 mg.

8. The method according to any one of the preceding claims, wherein the first antibody is administered at a dose of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, or about 1000 mg.

9. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously.

10. The method according to any one of the preceding claims, wherein the first antibody is administered by intravenous infusion over approximately 30 minutes.

11. The method according to any one of the preceding claims, wherein the first antibody is administered once a week.

12. The method according to any one of the preceding claims, wherein the first antibody is administered once every 2 weeks.

13. The method according to any one of the preceding claims, wherein the first antibody is administered once every 3 weeks.

14. The method according to any one of the preceding claims, wherein the first antibody is administered once every 4 weeks.

15. The method according to any one of the preceding claims, wherein the first antibody is administered once every 5 weeks.

16. The method according to any one of the preceding claims, wherein the first antibody is administered once every 6 weeks.

17. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 1 mg every 3 weeks.

18. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 3 mg every 3 weeks.

19. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 10 mg every 3 weeks.

20. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 30 mg every 3 weeks.

21. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 100 mg every 3 weeks.

22. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 300 mg once every 3 weeks.

23. The method according to any one of the preceding claims, wherein the first antibody is administered intravenously at a dose of 1000 mg once every 3 weeks.

24. The method according to any one of the preceding claims, wherein the cancer is a solid tumor.

25. The method of claim 24, wherein the solid tumor is metastatic or locally advanced.

26. The method according to claim 24 or 25, wherein the cancer is selected from the group consisting of: bladder cancer, brain cancer, breast cancer, cervical cancer, colon cancer and / or rectal cancer, endometrial cancer, head and neck cancer, kidney cancer, liver cancer, melanoma, mesothelioma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer, small cell lung cancer, gastric cancer, and thyroid cancer.

27. The method according to any one of the preceding claims, wherein before applying the first antibody: (a) The subject is at least 18 years old; (b) The subject was histologically diagnosed with a solid tumor that is currently metastatic or locally advanced, in which no standard therapy is available or standard therapy has failed; (c) The subject had measurable disease on baseline imaging, as assessed according to RECIST version 1.1; (d) The life expectancy of the subject > 3 months; (e) The subjects’ Eastern Oncology Collaboration Group performance status score was 0-1; (f) The subject has sufficient organ and bone marrow reserve function, as defined by one or more of the following: (i) Absolute neutrophil count > 1.5×10 9 / L; (ii) Platelet count > 100×10 9 / L; (iii) Hemoglobin > 8.0 g / dL, and no blood transfusion within 2 weeks prior to the hemoglobin measurement; (iv) serum albumin > 3.0 g / dL; (v) Total bilirubin < 1.5 × Upper limit of normal (ULN); (vi) Aspartate aminotransferase < 2.5×ULN; (vii) Alanine aminotransferase < 2.5×ULN; (viii) Alkaline phosphatase < 2.5×ULN, or for subjects with liver metastases < 5.0×ULN; (ix) Creatinine clearance > 40 mL / min, as assessed by the Cockcroft-Gault method; and / or (x) International Normalized Ratio or Prothrombin Time < 1.5×ULN and activation of partial thromboplastin time < 1.5×ULN, unless the patient is receiving anticoagulation therapy; (g) The subject is not pregnant and / or not breastfeeding; (h) The subjects mentioned above had not received cytotoxic therapy or other monoclonal antibody therapy within 3 weeks prior to enrollment; (i) The subjects had not received central nervous system (CNS) radiation therapy within 2 weeks prior to enrollment or non-CNS radiation therapy within 1 week prior to enrollment; and / or (j) The subjects in question had not received small molecule targeted therapy or tyrosine kinase inhibitor therapy within 2 weeks or 5 half-lives prior to enrollment; (k) The subject did not have any persistent toxicity from previous cancer therapies (Common Terminology Standard for Adverse Events [CTCAE]). > Level 1); (l) The subjects did not have sensory neuropathy or hair loss of grade >2; (m) The subjects described do not have a history of interstitial lung disease or active interstitial lung disease; (n) The subjects in question do not have a history of allergic reactions or uncontrolled asthma; (o) The subjects did not have any medical conditions requiring systemic treatment with corticosteroids (daily prednisone equivalent >10 mg) within 2 weeks prior to enrollment or with other immunosuppressive drugs within 30 days prior to enrollment; (p) The subject does not have active CNS metastases unless the CNS metastases have been treated and the subject is in a stable state both radiologically and clinically; (q) The subject does not have an active autoimmune disease or a history of autoimmune disease that requires systemic treatment within 2 years prior to enrollment, unless the treatment includes hormone replacement therapy or local treatment. (r) The subject has not received allogeneic tissue or solid organ transplantation, except for corneal transplantation; (s) The subjects did not have any active infection requiring treatment within 2 weeks prior to enrollment; (t) The subject is not HIV positive unless the subject's CD4 count is >200 cells / mm². 3 Or the viral load is undetectable; (u) The subject does not have active hepatitis B (HBV) or hepatitis C (HPC) unless hepatitis B surface antigen and HBV DNA or HCV RNA are negative; (v) The subject does not have clinically significant cardiovascular disease, as defined by one or more of the following: (i) Having experienced a cerebrovascular accident, stroke, or myocardial infarction within 6 months prior to enrollment; (ii) Unstable angina; (iii) Congestive heart failure (New York Heart Association classification) > Level II); and / or (iv) Uncontrolled arrhythmias requiring medication; (w) The subject's QTc interval is not greater than 480 milliseconds, unless the QTc prolongation is due to right bundle branch block; and / or (x) The subjects in question do not have uncontrolled hypertension (>140 / 90 mmHg) or hypertension that requires more than 3 antihypertensive medications to control.

28. The method according to any one of the preceding claims, wherein the first antibody comprises the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 as shown in SEQ ID NO: 1, 2, 3, 4, 5 and 6, respectively.

29. The method according to any one of the preceding claims, wherein the first antibody comprises the VH amino acid sequence of SEQ ID NO: 7 and / or the VL amino acid sequence of SEQ ID NO:

8.

30. The method according to any one of the preceding claims, wherein the first antibody comprises the VH amino acid sequence and the VL amino acid sequence, respectively, SEQ ID NO: 7 and 8.

31. The method according to any one of the preceding claims, wherein the first antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 and IgM.

32. The method of claim 31, wherein the heavy chain constant region is the human IgG4 heavy chain constant region, the human IgG4 heavy chain constant region comprising proline at position 228 according to the EU numbering system.

33. The method according to any one of the preceding claims, wherein the first antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 9; and / or a light chain comprising the amino acid sequence of SEQ ID NO:

10.

34. The method according to any one of the preceding claims, wherein the first antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 9 and 10, respectively.

35. The method according to any one of claims 2 or 6 to 32, wherein the human PD-1 inhibitor is a second antibody that specifically binds to human PD-1, human PD-L1 or human PD-L2.

36. The method of claim 35, wherein the second antibody comprises: VH, the VH comprising the CDRH1, CDRH2 and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO:17; and VL, the VL comprising the CDRL1, CDRL2 and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO:

18.

37. The method according to claim 35 or 36, wherein the second antibody comprises the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 as shown in SEQ ID NO: 11, 12, 13, 14, 15 and 16, respectively.

38. The method according to any one of claims 35 or 36, wherein the second antibody comprises the VH amino acid sequence of SEQ ID NO: 17 and / or the VL amino acid sequence of SEQ ID NO:

18.

39. The method according to any one of claims 35 to 38, wherein the second antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 17 and 18, respectively.

40. The method according to any one of claims 35 to 39, wherein the second antibody comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM.

41. The method of claim 40, wherein the heavy chain constant region is the human IgG4 heavy chain constant region, the human IgG4 heavy chain constant region comprising proline at position 228 according to the EU numbering system.

42. The method according to any one of claims 35 to 41, wherein the second antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 19; and / or a light chain comprising the amino acid sequence of SEQ ID NO:

20.

43. The method according to any one of claims 35 to 42, wherein the second antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 19 and 20, respectively.

44. The method of claim 35, wherein the second antibody is selected from the group consisting of: batitilimab, adebenone, atezolizumab, averuzumab, camrelizumab, simipremab, cochilizumab, dostalimumab, durvalumab, enronsubab, envorizumab, nivolumab, pembrolizumab, pendilizumab, palolizumab, pectilizumab, pravalimumab, slulizumab, sintilimumab, socazolimumab, sugalizumab, tagorizumab, tislelizumab, toripalimab, and cepalimumab.

45. The method according to any one of claims 35 to 44, wherein the second antibody is administered at a dose of about 450 mg.

46. ​​The method according to any one of claims 35 to 44, wherein the second antibody is administered at a dose of about 3 mg / kg.

47. The method according to any one of claims 35 to 46, wherein the second antibody is administered by intravenous infusion over about 30 minutes.

48. The method according to any one of claims 35 to 47, wherein the second antibody is administered once a week.

49. The method according to any one of claims 35 to 47, wherein the second antibody is administered once every 2 weeks.

50. The method according to any one of claims 35 to 47, wherein the second antibody is administered once every 3 weeks.

51. The method according to any one of claims 35 to 50, wherein the second antibody is administered to the subject after the first antibody is administered.

52. The method according to any one of claims 3 and 6 to 51, wherein the human CTLA-4 inhibitor is a third antibody that specifically binds to human CTLA-4.

53. The method of claim 52, wherein the third antibody comprises: VH, the VH comprising the CDRH1, CDRH2 and CDRH3 amino acid sequences of the VH amino acid sequence shown in SEQ ID NO:27; and VL, the VL comprising the CDRL1, CDRL2 and CDRL3 amino acid sequences of the VL amino acid sequence shown in SEQ ID NO:

28.

54. The method according to claim 52 or 53, wherein the third antibody comprises the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 as shown in SEQ ID NO: 21, 22, 23, 24, 25 and 26, respectively.

55. The method according to any one of claims 52 to 54, wherein the third antibody comprises the VH amino acid sequence of SEQ ID NO: 27 and / or the VL amino acid sequence of SEQ ID NO:

28.

56. The method according to any one of claims 52 to 55, wherein the third antibody comprises the VH amino acid sequence and the VL amino acid sequence of SEQ ID NO: 27 and 28, respectively.

57. The method according to any one of claims 52 to 56, wherein the third antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 and IgM.

58. The method according to any one of claims 52 to 57, wherein the third antibody comprises a human IgG1 heavy chain constant region, the heavy chain constant region being a variant of the wild-type human IgG1 heavy chain constant region, wherein the binding affinity of the variant human IgG1 heavy chain constant region to FcγRIIIA is higher than that of the wild-type human IgG1 heavy chain constant region to FcγRIIIA.

59. The method of claim 58, wherein the third antibody is unfucosylated.

60. The method according to any one of claims 57 to 59, wherein the heavy chain constant region is the human IgG1 heavy chain constant region, the human IgG1 heavy chain constant region comprising aspartic acid at position 239, leucine at position 330, and glutamic acid at position 332 according to the EU numbering system.

61. The method according to any one of claims 52 to 60, wherein the third antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 29; and / or a light chain comprising the amino acid sequence of SEQ ID NO:

30.

62. The method according to any one of claims 52 to 61, wherein the third antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 29 and 30, respectively.

63. The method according to any one of claims 52 to 53, wherein the third antibody is selected from the group consisting of: botelimab, ipilimumab, and trimemumab.

64. The method according to any one of claims 52 to 63, wherein the third antibody is administered at a dose of about 150 mg.

65. The method according to any one of claims 52 to 63, wherein the third antibody is administered at a dose of about 75 mg.

66. The method according to any one of claims 52 to 63, wherein the third antibody is administered at a dose of about 50 mg.

67. The method according to any one of claims 52 to 63, wherein the third antibody is administered at a dose of about 2 mg / kg.

68. The method according to any one of claims 52 to 63, wherein the third antibody is administered at a dose of about 4 mg / kg.

69. The method according to any one of claims 52 to 68, wherein the third antibody is administered by intravenous infusion over approximately 30 minutes.

70. The method according to any one of claims 52 to 69, wherein the third antibody is administered once every 2 weeks.

71. The method according to any one of claims 52 to 69, wherein the third antibody is administered once every 3 weeks.

72. The method according to any one of claims 52 to 69, wherein the third antibody is administered once every 4 weeks.

73. The method according to any one of claims 52 to 69, wherein the third antibody is administered once every 6 weeks.

74. The method according to any one of claims 52 to 73, wherein the third antibody is administered to the subject after the first antibody is administered.

75. The method according to any one of claims 52 to 73, wherein the third antibody is administered to the subject after the administration of the first antibody and the second antibody.

76. The method according to any one of claims 2 or 6 to 34, wherein the human PD-1 inhibitor comprises an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

77. The method according to any one of claims 3 or 6 to 34, wherein the human CTLA-4 inhibitor comprises an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

78. The method according to any one of claims 4 or 7 to 34, wherein the human PD-L1 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

79. The method according to any one of claims 5 or 7 to 34, wherein the human PD-L2 inhibitor comprises an antagonistic antibody or a fragment thereof, an inhibitory small molecule, an inhibitory peptide, or an inhibitory Fc fusion protein.

80. A method of treating cancer in a subject in need, the method comprising administering to the subject: (a) A first antibody that specifically binds to human ILT2, wherein the antibody comprises: a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 9 and 10, respectively; (b) Batelimab administered intravenously at a dose of 450 mg every 3 weeks; and (c) Bortelimab administered via intravenous infusion at a dose of 150 mg every 6 weeks.

81. The method of claim 80, wherein the first antibody is administered at a dose of about 1 mg to about 1000 mg.

82. The method according to claim 80 or 81, wherein the first antibody is administered at a dose of about 1 mg, about 3 mg, about 10 mg, about 30 mg, about 100 mg, about 300 mg, or about 1000 mg.

83. The method according to any one of claims 80 to 82, wherein the first antibody is administered intravenously.

84. The method according to any one of claims 80 to 83, wherein the first antibody is administered by intravenous infusion over about 30 minutes.

85. The method according to any one of claims 80 to 84, wherein the first antibody is administered once a week.

86. The method according to any one of claims 80 to 84, wherein the first antibody is administered once every 2 weeks.

87. The method according to any one of claims 80 to 84, wherein the first antibody is administered once every 3 weeks.

88. The method according to any one of claims 80 to 84, wherein the first antibody is administered once every 4 weeks.

89. The method according to any one of claims 80 to 84, wherein the first antibody is administered once every 5 weeks.

90. The method according to any one of claims 80 to 84, wherein the first antibody is administered once every 6 weeks.

91. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 1 mg every 3 weeks.

92. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 3 mg every 3 weeks.

93. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 10 mg every 3 weeks.

94. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 30 mg every 3 weeks.

95. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 100 mg every 3 weeks.

96. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 300 mg every 3 weeks.

97. The method according to any one of claims 80 to 84, wherein the first antibody is administered intravenously at a dose of 1000 mg every 3 weeks.

98. The method according to any one of claims 80 to 97, wherein the cancer is a solid tumor.

99. The method of claim 98, wherein the solid tumor is metastatic or locally advanced.

100. The method according to claim 98 or 99, wherein the cancer is selected from the group consisting of: bladder cancer, brain cancer, breast cancer, cervical cancer, colon cancer and / or rectal cancer, endometrial cancer, head and neck cancer, kidney cancer, liver cancer, melanoma, mesothelioma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer, small cell lung cancer, gastric cancer, and thyroid cancer.

101. An antibody that specifically binds to human ILT2 for use in the treatment of cancer, wherein the treatment is performed according to the method of any one of the preceding claims.

102. An antibody that specifically binds to human ILT2 for use in the manufacture of a medicament for treating cancer, wherein the treatment is performed according to the method of any one of the preceding claims.

103. Use of an antibody that specifically binds to human ILT-2 for the treatment of cancer, wherein the treatment is performed according to the method of any one of the preceding claims.

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