Cancer Therapy Targeting NKG2A

Abstract

The present invention relates to anti-NKG2A antibodies, optionally in combination with anti-PD-1 or anti-PD-L1 antibodies and / or anti-EGFR or anti-HER2 antibodies, and methods of using said antibodies or antibody combinations in enhancing immunity and in the treatment of cancer in patients in need thereof.

Description

[Technical field]

[0001] 2. Background of the Invention Cancer is ranked as the leading cause of death and a significant barrier to increasing life expectancy in every country in the world. According to World Health Organization (WHO) estimates in 2019, cancer is the first or second cause of death before the age of 70 in 112 of 183 countries, and third or fourth in a further 23 countries.

[0002] HER2-overexpressing gastric cancer is an area of ​​unmet medical need with limited treatment options. Amplification of the HER2 (also known as ERBB2) oncogene and overexpression of the HER2 protein occurs in approximately 17-20% of patients with gastric cancer. Patients with HER2-overexpressing gastric cancer benefit from treatment with the anti-HER2 antibody trastuzumab in combination with cisplatin and 5FU or capecitabine in the first line; however, after progression on trastuzumab-based therapy, options are limited for treating HER2+ advanced gastric cancer.

[0003] Treatment for metastatic colorectal cancer (mCRC) is heavily dependent on gene expression, with many drugs only prescribed for specific tumor profiles. Anti-PD-1 agents, such as nivolumab and pembrolizumab, have seen early success in microsatellite instability-high / deficient mismatch repair (MSI-H / dMMR) mCRC, but these comprise only 15% of patients. RAS / BRAF wild-type (WT) and EGFR-positive tumors are among the most common gene expression profiles, accounting for approximately 40% of CRCs.

[0004] NKG2A (CD159a) is a C-type lectin that heterodimerizes with CD94 to create an immunoinhibitory receptor expressed on natural killer (NK) cells, NKT cells, gamma delta (γδ) T cells, and a subset of cytotoxic T cells (Borrego et al., Immunol Res (2006) 35(3):263-78; Vivier et al., Nat Rev Immunol. (2004) 4(3):190-8). Upon ligation to its ligand, human leukocyte antigen (HLA)-E, NKG2A / CD94 transmits inhibitory signals via two immunotyrosine-based inhibitory motifs in its cytoplasmic tail and recruitment of the SHP-1 tyrosine phosphotase (Carotta et al., Front Immunol. (2016) 7:152). This mechanism is part of the natural self-recognition / tolerance by NK cells. However, cancer cells exploit this system by overexpressing HLA-E, thereby protecting themselves against NK cell and T cell-mediated killing. In patients, NKG2A expression is increased on tumor-infiltrating NK cells and T cells and can be induced by immunosuppressive factors such as TGF-β and adenosine (Platonova et al., Cancer Res.(2011)71(16):5412-22;Sheu et al., Cancer Res.(2005)65(7):2921-9). In fact, high intratumoral expression of NKG2A and HLA-E predicts poor prognosis for patients with hepatocellular carcinoma (HCC) (Sun et al., Oncoimmunology(2017)6(1):e1264562). Summary of the Invention [Problem to be solved by the invention]

[0005] Given the critical roles of NKG2A, PD-1 and PD-L1, HER2, and EGFR in cancer, there is a need for new and improved therapies that target these receptors (e.g., in combination) to treat cancer. [Means for solving the problem]

[0006] Summary of the Invention The present invention is based on a treatment for enhancing immunity, comprising, for example, an anti-NKG2A antibody as described herein, optionally, for example, an antibody targeting PD-1 or PD-L1 as described herein, and / or an antibody targeting EGFR or HER2. In some embodiments, the treatment is for treating cancer. Also provided are pharmaceutical compositions comprising the components of the treatment, and the use of the treatment for enhancing immunity in a patient (e.g., treating cancer). The treatment described herein may be used in a method for enhancing immunity in a patient (e.g., treating cancer); may be used for the manufacture of a medicament for enhancing immunity in a patient (e.g., treating cancer); or may be used to enhance immunity in a patient (e.g., treating cancer). It is contemplated that the treatment described herein may provide superior clinical responses compared to currently available treatments (e.g., for cancer), including antibody treatments.

[0007] In some embodiments, the present disclosure provides a method of enhancing immunity in a human patient in need thereof, comprising administering to the patient: a) an anti-NKG2A antibody or antigen-binding portion thereof that competes or cross-competes for binding to human NKG2A or binds to the same epitope of human NKG2A as an antibody comprising the heavy and light chain amino acid sequences of SEQ ID NOs: 9 and 10, respectively, or SEQ ID NOs: 19 and 20, respectively; and, optionally b) an anti-PD-1 antibody or an anti-PD-L1 antibody, or an antigen-binding portion thereof; and optionally c) An anti-EGFR antibody component ("anti-EGFR component"), which comprises one or two anti-EGFR antibodies or antigen-binding portions thereof, or an anti-HER2 antibody.

[0008] In certain embodiments, the method comprises administering: a) an anti-NKG2A antibody, or an antigen-binding portion thereof, and an anti-PD-1 antibody, or an antigen-binding portion thereof; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, and an anti-PD-L1 antibody, or an antigen-binding portion thereof; c) an anti-NKG2A antibody or an antigen-binding portion thereof, an anti-PD-1 antibody or an antigen-binding portion thereof, and an anti-EGFR component; d) an anti-NKG2A antibody or an antigen-binding portion thereof, an anti-PD-1 antibody or an antigen-binding portion thereof, and an anti-HER2 antibody or an antigen-binding portion thereof; e) an anti-NKG2A antibody or antigen-binding portion thereof, an anti-PD-L1 antibody or antigen-binding portion thereof, and an anti-EGFR component; or f) An anti-NKG2A antibody or an antigen-binding portion thereof, an anti-PD-L1 antibody or an antigen-binding portion thereof, and an anti-HER2 antibody or an antigen-binding portion thereof.

[0009] In some embodiments, heavy chain complementarity determining regions (H-CDRs) 1 to 3 and light chain complementarity determining regions (L-CDRs) 1 to 3 of the anti-NKG2A antibody comprise the amino acid sequences of SEQ ID NOs: 1 to 6, respectively; or SEQ ID NOs: 11 to 16, respectively. In specific embodiments, the heavy chain variable domain (VH) and light chain variable domain (VL) of the anti-NKG2A antibody comprise the amino acid sequences of SEQ ID NOs: 7 and 8, respectively; or SEQ ID NOs: 17 and 18, respectively. In specific embodiments, the heavy chain (HC) and light chain (LC) of the anti-NKG2A antibody comprise the amino acid sequences of SEQ ID NOs: 9 and 10, respectively; or SEQ ID NOs: 19 and 20, respectively.

[0010] In some embodiments, the H-CDR1-3 and L-CDR1-3 of the anti-PD-1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 21 to 26, respectively; b) SEQ ID NOs: 31 to 36, respectively; c) SEQ ID NOs: 41 to 46, respectively; d) SEQ ID NOs: 51 to 56, respectively; e) SEQ ID NOs: 61 to 66, respectively; or f) SEQ ID NOs: 71 to 76, respectively.

[0011] In certain embodiments, the VH and VL of the anti-PD-1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 27 and 28, respectively; b) SEQ ID NOs: 37 and 38, respectively; c) SEQ ID NOs: 47 and 48, respectively; d) SEQ ID NOs: 57 and 58, respectively; e) SEQ ID NOs: 67 and 68, respectively; or f) SEQ ID NOs: 77 and 78, respectively.

[0012] In certain embodiments, the HC and LC of the anti-PD-1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 29 and 30, respectively; b) SEQ ID NOs: 39 and 40, respectively; c) SEQ ID NOs: 49 and 50, respectively; d) SEQ ID NOs: 59 and 60, respectively; e) SEQ ID NOs: 69 and 70, respectively; or f) SEQ ID NOs: 79 and 80, respectively.

[0013] The anti-PD-1 antibody can be, for example, nivolumab, pembrolizumab, cemiplimab, dostallimab, or retifanlimab.

[0014] In some embodiments, the H-CDR1-3 and L-CDR1-3 of the anti-PD-L1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 81 to 86, respectively; b) SEQ ID NOs: 91 to 96, respectively; or c) SEQ ID NOs: 101 to 106, respectively.

[0015] In certain embodiments, the VH and VL of the anti-PD-L1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 87 and 88, respectively; b) SEQ ID NOs: 97 and 98, respectively; or c) SEQ ID NOs: 107 and 108, respectively.

[0016] In certain embodiments, the HC and LC of the anti-PD-L1 antibody comprise the following amino acid sequences: a) SEQ ID NOs: 89 and 90, respectively; b) SEQ ID NOs: 99 and 100, respectively; or c) SEQ ID NOs: 109 and 110, respectively.

[0017] The anti-PD-L1 antibody can be, for example, atezolizumab, avelumab, or durvalumab.

[0018] In some embodiments, the anti-EGFR component comprises an anti-EGFR antibody, or antigen-binding portion thereof, having heavy chain complementarity determining regions (H-CDRs) 1-3 and light chain complementarity determining regions (L-CDRs) 1-3 comprising the following amino acid sequences: a) SEQ ID NOs: 111 to 116, respectively; b) SEQ ID NOs: 121 to 126, respectively; c) SEQ ID NOs: 131 to 136, respectively; or d) SEQ ID NOs: 141 to 146, respectively.

[0019] In certain embodiments, the anti-EGFR antibody, or antigen-binding portion thereof, comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 117 and 118, respectively; b) SEQ ID NOs: 127 and 128, respectively; c) SEQ ID NOs: 137 and 138, respectively; or d) SEQ ID NOs: 147 and 148, respectively.

[0020] In certain embodiments, the anti-EGFR antibody comprises a HC and a LC comprising the following amino acid sequences: a) SEQ ID NOs: 119 and 120, respectively; b) SEQ ID NOs: 129 and 130, respectively; c) SEQ ID NOs: 139 and 140, respectively; or d) SEQ ID NOs: 149 and 150, respectively.

[0021] The anti-EGFR antibody can be, for example, cetuximab, panitumumab, futuximab, or modotuximab.

[0022] In some embodiments, the anti-EGFR component comprises an anti-EGFR antibody or antigen-binding portion thereof having H-CDR1-3 and L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 131-136, respectively, and an anti-EGFR antibody or antigen-binding portion thereof having H-CDR1-3 and L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 141-146, respectively. In certain embodiments, the anti-EGFR component comprises an anti-EGFR antibody or antigen-binding portion thereof having VH and VL comprising the amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an anti-EGFR antibody or antigen-binding portion thereof having VH and VL comprising the amino acid sequences of SEQ ID NOs: 147 and 148, respectively. In certain embodiments, the anti-EGFR component comprises an anti-EGFR antibody having heavy chain (HC) and light chain (LC) comprising the amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody having HC and LC comprising the amino acid sequences of SEQ ID NOs: 149 and 150, respectively. The anti-EGFR components can be, for example, futuximab plus modotuximab (eg, in a 1:1 ratio).

[0023] In some embodiments, H-CDR1-3 and L-CDR1-3 of the anti-HER2 antibody comprise the amino acid sequences of SEQ ID NOs: 151-156, respectively, or SEQ ID NOs: 161-166, respectively. In certain embodiments, VH and VL of the anti-HER2 antibody comprise the amino acid sequences of SEQ ID NOs: 157 and 158, respectively, or SEQ ID NOs: 167 and 168, respectively. In certain embodiments, HC and LC of the anti-HER2 antibody comprise the amino acid sequences of SEQ ID NOs: 159 and 160, respectively, or SEQ ID NOs: 169 and 170, respectively. In some embodiments, the anti-HER2 antibody or antigen-binding portion may be conjugated to a moiety, such as DXd or DM1. The anti-HER2 antibody may be, for example, trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine.

[0024] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; and an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 61 to 66, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; and an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 67 and 68, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; and an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively.

[0025] In some embodiments, the method comprises administering to the patient: an anti-NKG2A antibody or an antigen-binding portion thereof comprising the amino acid sequences of H-CDR1 to 3 and L-CDR1 to 3 of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 61 to 66, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; an anti-NKG2A antibody or an antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 67 and 68, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and Anti-EGFR components include an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0026] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 61 to 66, respectively; and an anti-HER2 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 161 to 166, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 67 and 68, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 167 and 168, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0027] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 71 to 76, respectively; and an anti-HER2 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 161 to 166, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 77 and 78, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 167 and 168, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 79 and 80, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0028] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and an anti-HER2 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 151 to 156, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 157 and 158, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:39 and 40, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:159 and 160, respectively.

[0029] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and an anti-HER2 antibody or antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 151 to 156, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 157 and 158, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:29 and 30, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:159 and 160, respectively.

[0030] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the amino acid sequences of H-CDR1 to 3 and L-CDR1 to 3 of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; b) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; and Anti-EGFR components include an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150.

[0031] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the amino acid sequences of H-CDR1 to 3 and L-CDR1 to 3 of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; b) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-EGFR component comprising an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an anti-EGFR antibody or antigen-binding portion thereof comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and Anti-EGFR components include an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150.

[0032] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 111 to 116, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-EGFR antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 117 and 118, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:39 and 40, respectively; and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:119 and 120, respectively.

[0033] In some embodiments, the method comprises administering to the patient: a) an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 111 to 116, respectively; b) an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-EGFR antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 117 and 118, respectively; or c) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:29 and 30, respectively; and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs:119 and 120, respectively.

[0034] In any of the above methods, the antibodies or antigen-binding portions may be administered to the patient simultaneously or sequentially.

[0035] In some embodiments, the patient has cancer, e.g., a hematological malignancy or a solid tumor, hi certain embodiments, the cancer is colorectal cancer or gastric cancer.

[0036] In some embodiments, the anti-NKG2A antibody or antigen-binding portion thereof is administered at a dose of 8, 20, 100, 300, 750, or 1500 mg (e.g., every 2 weeks). The antibody or antigen-binding portion may be administered in a 28-day cycle. In some embodiments, the anti-PD-1 or anti-PD-L1 antibody or antigen-binding portion thereof is administered at a dose of 200 mg (e.g., every 2 weeks), and in some cases may be administered after one cycle of the anti-NKG2A antibody or antigen-binding portion. In some embodiments, the anti-EGFR component is administered at a dose of 6 mg / kg, 9 mg / kg, or 9 mg / kg followed by a loading dose of 6 mg / kg (e.g., every week or every 2 weeks). In some embodiments, the anti-HER2 antibody or antigen-binding portion thereof is administered at a dose of 15 mg / kg (e.g., every 3 weeks or every 4 weeks). In certain embodiments, the antibody or antigen-binding portion is formulated for intravenous administration (e.g., intravenous infusion).

[0037] The present disclosure provides a method of treating progressive solid tumor malignancies in a patient comprising administering to the patient an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively, for example, where the antibody is administered by IV infusion every two weeks at doses of 8, 20, 100, 300, 750, or 1500 mg.

[0038] The present disclosure further provides a method of treating an advanced solid tumor malignancy in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; and b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; For example, wherein the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg by IV infusion every two weeks, and wherein, after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg by IV infusion every two weeks.

[0039] The present disclosure further provides a method of treating an advanced solid tumor malignancy in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; and b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; For example, wherein the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg by IV infusion every two weeks, and wherein, after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg by IV infusion every two weeks.

[0040] The present disclosure further provides a method of treating an advanced solid tumor malignancy in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; and b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; For example, wherein the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg by IV infusion every two weeks, and wherein, after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg by IV infusion every two weeks.

[0041] The present disclosure also provides a method for treating metastatic HER2 + A method of treating gastric cancer is provided, comprising administering to a patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and c) An anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0042] In some embodiments, the cancer is locally advanced and / or unresectable. In some embodiments, the patient has failed first-line standard of care therapy. In certain embodiments, the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg every 2 weeks; after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg every 2 weeks; and the anti-HER2 antibody is administered at 15 mg / kg every 3 or 4 weeks, where the antibodies are administered via IV infusion.

[0043] The present disclosure also provides a method for treating metastatic HER2 + A method of treating gastric cancer is provided, comprising administering to a patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 79 and 80, respectively; and c) An anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0044] In some embodiments, the cancer is locally advanced and / or unresectable. In some embodiments, the patient has failed first-line standard of care therapy. In certain embodiments, the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg every 2 weeks; after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg every 2 weeks; and the anti-HER2 antibody is administered at 15 mg / kg every 3 or 4 weeks, where the antibodies are administered via IV infusion.

[0045] The disclosure also provides a method of treating metastatic colorectal cancer in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and c) An anti-EGFR component, comprising an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150.

[0046] In some embodiments, the patient has a low microsatellite instability status. In some embodiments, the patient (i) does not have a RAS mutation in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4; and / or (ii) does not have a BRAF V600E mutation. In certain embodiments, the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg every two weeks; after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg every two weeks; and the anti-EGFR component is administered at a loading dose of 9 mg / kg followed by a dose of 6 mg / kg every one or two weeks, where the antibodies are administered via IV infusion.

[0047] The disclosure also provides a method of treating metastatic colorectal cancer in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; and c) An anti-EGFR component, comprising an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0048] In some embodiments, the patient has a low microsatellite instability status. In some embodiments, the patient (i) does not have a RAS mutation in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4; and / or (ii) does not have a BRAF V600E mutation. In certain embodiments, the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg every two weeks; after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg every two weeks; and the anti-EGFR component is administered at a loading dose of 9 mg / kg followed by a dose of 6 mg / kg every one or two weeks, where the antibodies are administered via IV infusion.

[0049] The disclosure also provides a method of treating metastatic colorectal cancer in a patient, comprising administering to the patient: a) an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; b) an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and c) An anti-EGFR component, comprising an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150.

[0050] In some embodiments, the patient has a low microsatellite instability status. In some embodiments, the patient (i) does not have a RAS mutation in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4; and / or (ii) does not have a BRAF V600E mutation. In certain embodiments, the anti-NKG2A antibody is administered at a dose of 20, 100, 300, 750, or 1500 mg every two weeks; after completing a 28-day cycle of anti-NKG2A antibody administration, the anti-PD-1 antibody is administered at a dose of 200 mg every two weeks; and the anti-EGFR component is administered at a loading dose of 9 mg / kg followed by a dose of 6 mg / kg every one or two weeks, where the antibodies are administered via IV infusion.

[0051] In some embodiments, the methods of the present disclosure further comprise administering to the patient radiation therapy, or at least one of a chemotherapeutic agent, an anti-tumor agent, and an anti-angiogenic agent.

[0052] Treatment with the methods of the present disclosure may result in tumor regression, slowing of tumor progression, inhibition of cancer progression, inhibition of cancer metastasis, prevention of cancer recurrence or residual disease, and / or extended survival. In some embodiments, treatment with the methods of the present disclosure results in improved objective response rate, improved clinical benefit rate, improved duration of response, increased progression-free survival, increased overall survival, or any combination thereof, e.g., in comparison to untreated patients.

[0053] The present disclosure also provides multispecific antibodies that specifically bind to: a) human NKG2A and human PD-1; b) human NKG2A and human PD-L1; c) human NKG2A, human PD-1, and human EGFR; d) human NKG2A, human PD-1, and human HER2; e) human NKG2A, human PD-L1, and human EGFR; or f) Human NKG2A, human PD-L1, and human HER2.

[0054] In some embodiments, the multispecific antibody comprises: a) the antigen-binding domain of an anti-NKG2A antibody described herein, and the antigen-binding domain of an anti-PD-1 antibody described herein; b) the antigen-binding domain of an anti-NKG2A antibody described herein, and the antigen-binding domain of an anti-PD-L1 antibody described herein; c) an antigen-binding domain of an anti-NKG2A antibody described herein, an antigen-binding domain of an anti-PD-1 antibody described herein, and an antigen-binding portion of one or two anti-EGFR antibodies described herein; d) an antigen-binding domain of an anti-NKG2A antibody described herein, an antigen-binding domain of an anti-PD-1 antibody described herein, and an antigen-binding portion of an anti-HER2 antibody described herein; e) the antigen-binding domain of an anti-NKG2A antibody described herein, the antigen-binding domain of an anti-PD-L1 antibody described herein, and the antigen-binding portions of one or two anti-EGFR antibodies described herein; or f) The antigen-binding domain of an anti-NKG2A antibody described herein, the antigen-binding domain of an anti-PD-L1 antibody described herein, and the antigen-binding portion of an anti-HER2 antibody described herein.

[0055] The present disclosure also provides pharmaceutical compositions comprising an anti-NKG2A antibody, or an antigen-binding portion thereof, described herein, and further comprising: a) an anti-PD-1 antibody, or an antigen-binding portion thereof; b) an anti-PD-L1 antibody, or an antigen-binding portion thereof; c) an anti-PD-1 antibody or antigen-binding portion thereof, and an anti-EGFR component; d) an anti-PD-1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof; e) an anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-EGFR component; or f) an anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof; and a pharma- ceutical acceptable excipient. The anti-PD-1 antibody or antigen-binding portion thereof, the anti-PD-L1 antibody or antigen-binding portion thereof, the anti-EGFR component, and the anti-HER2 antibody or antigen-binding portion thereof can be, for example, as described herein. In certain embodiments, the pharmaceutical composition can include an antibody or antigen-binding portion of any of the methods described herein, and can be for use in treating a human patient in any of the methods described herein.

[0056] The disclosure also provides an anti-NKG2A antibody, or antigen-binding portion thereof, described herein, for use in boosting immunity in a human patient in need thereof, in combination with: a) an anti-PD-1 antibody, or an antigen-binding portion thereof; b) an anti-PD-L1 antibody, or an antigen-binding portion thereof; c) an anti-PD-1 antibody or antigen-binding portion thereof, and an anti-EGFR component; d) an anti-PD-1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof; e) an anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-EGFR component; or f) An anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof.

[0057] The anti-PD-1 antibody or antigen-binding portion thereof, the anti-PD-L1 antibody or antigen-binding portion thereof, the anti-EGFR component, and the anti-HER2 antibody or antigen-binding portion thereof can be, for example, as described herein. In some embodiments, the anti-NKG2A antibody or antigen-binding portion thereof is for use in treating a human patient in the methods described herein.

[0058] The disclosure also provides for the use of an anti-NKG2A antibody, or antigen-binding portion thereof, described herein in combination with: a) an anti-PD-1 antibody, or an antigen-binding portion thereof; b) an anti-PD-L1 antibody, or an antigen-binding portion thereof; c) an anti-PD-1 antibody or antigen-binding portion thereof, and an anti-EGFR component; d) an anti-PD-1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof; e) an anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-EGFR component; or f) An anti-PD-L1 antibody, or an antigen-binding portion thereof, and an anti-HER2 antibody, or an antigen-binding portion thereof. For the manufacture of a medicament for enhancing immunity in a human patient in need thereof. The anti-PD-1 antibody or antigen-binding portion thereof, the anti-PD-L1 antibody or antigen-binding portion thereof, the anti-EGFR component, and the anti-HER2 antibody or antigen-binding portion thereof can be, for example, as described herein. In some embodiments, the medicament is for treating a human patient in the methods described herein.

[0059] Other features, objects, and advantages of the present invention will become apparent in the detailed description which follows. It should be understood, however, that the detailed description, while indicating embodiments and aspects of the present invention, is given by way of illustration only, and not by way of limitation. Various changes and modifications within the scope of the present invention will become apparent to those skilled in the art from the detailed description. [Brief description of the drawings]

[0060] [Figure 1] FIG. 1 is a pair of graphs showing the expression of endogenous HLA-E on the surface of six different tumor cell lines (Panel A) and the effect of mAb1 on NK-mediated killing of these six tumor cell lines (Panel B). [Diagram 2] FIG. 2 is a pair of graphs showing γδ T cell-mediated tumor cell killing by mAb1 compared to a monalizumab analog (Panel A) and an analog of the BMS anti-NKG2A mAb (Panel B) in vitro. One representative donor is depicted; results were validated using n=3 donors. Data are presented as mean ± SEM. [Diagram 3]FIG. 3 is a series of graphs showing the percentage specific lysis of FaDu cells (HLA-E+ / EGFR+) by cetuximab titrated in a dose-dependent manner, alone or in combination with mAb1, monalizumab analogs, or isotype control. [Figure 4A] Figure 4A is a pair of graphs showing mAb1-induced enhancement of NK cell-mediated killing of A431 tumor cells in combination with the anti-EGFR antibody cetuximab in two human donors (D1 and D2). Data are presented as mean ± SEM. [Figure 4B] Figure 4B is a pair of graphs showing mAb1-induced enhancement of NK cell-mediated killing of A431 tumor cells in combination with the anti-EGFR antibody combination futuximab / modotuximab ("Futux / Modo") in two human donors (D1 and D2). Data are presented as mean ± SEM. [Diagram 5] Figure 5 is a pair of graphs showing NK cell activation (as assessed by CD137 expression) affected by mAb1 alone or in combination with the anti-EGFR antibody cetuximab ("cetux") (panel A) or the anti-EGFR antibody combination futuximab / modotuximab ("futux / modo") (panel B). Each data point represents a donor. [Figure 6] 6 is a graph showing IFNγ secretion by primary NK cells in vitro in the presence of A431 cells, IL-2, and the indicated antibodies or antibody combinations. Each data point represents a donor. [Figure 7] Figure 7 is a pair of graphs showing mAb1-induced enhancement of NK cell-mediated killing of A431 or MDA-MB-231 tumor cells in combination with the anti-PD-L1 antibody avelumab in two human donors (D1 and D2). Data are presented as mean ± SEM. [Figure 8] FIG. 8 is a pair of graphs showing the effect of mAb1 in combination with avelumab ("Ave") on induction of NK cell activation (CD137) (Panel A) and IFNγ secretion (Panel B). [Figure 9A] Figure 9A is a graph showing tumor growth in CD34 humanized mice subcutaneously implanted with MDA-MB-231 human breast tumor cells. The grey area indicates the treatment period. Data are presented as mean ± SEM. *p<0.05. [Figure 9B] Figure 9B is a series of graphs showing flow cytometry analysis of MDA-MB-231 tumor-infiltrating lymphocytes. The percentage of human CD45+ and CD3+ cells and the ratio of CD8 / CD4 T cells are shown for tumors treated with the indicated antibodies or antibody combinations compared to vehicle. Numbers are presented as percentage of live cells (CD45+), human CD45+ cells (CD3+), or ratios (CD4 and CD8). [Figure 10] FIG. 10 shows the clinical trial design of mAb1 monotherapy and combination therapy. [Figure 11] Figure 11 shows the percentage of tumor eradication in hNKG2A / hCD94 KI mice subcutaneously implanted with MC38-HLAE mouse tumor cells. Mice were treated three times a week with a total of nine doses. Tumor volumes were measured three times a week. The grey area indicates the treatment period. Data are presented as mean ± SEM. [Figure 12] Figure 12 shows trastuzumab and mAb1-induced NK cell secretion of MIP-1β. Human primary NKG2A+ NK cells from healthy individuals were co-cultured with HLA-E-transduced N87, BxPC3, SKOV3, A375, A549 and JIMT-1 target cells for 48 hours in the presence of 10 ng / mL IL-2. Secretion of MIP-1β in the co-culture supernatants was quantified by ELISA. [Figure 13] Figure 13 is a series of graphs showing the combined effect of dual PD-1 and NKG2A blockade on peripheral blood lymphocytes (PBLs) isolated from three healthy donors (A, B, and C). PBLs were co-cultured with HER2-positive SKOV3 cells overexpressing HLA-E and PD-L1 and activated with trastuzumab or trastuzumab + zoledronate. [Figure 14]FIG. 14 shows the clinical trial design of mAb1 + pembrolizumab in the treatment of colorectal cancer. [Figure 15] FIG. 15 shows the design of a clinical trial of mAb1 + pembrolizumab + trastuzumab in the treatment of gastric cancer. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0061] Detailed Description of the Invention The present disclosure provides new monotherapies and compositions that target human NKG2A, as well as new combination therapies and compositions that target human NKG2A; human PD-1 or PD-L1; and / or human EGFR or HER2 by using antibodies that bind to these targets. The treatments (i.e., monotherapies and combination therapies) and compositions can be used to treat cancer in human patients. Unless otherwise stated, "NKG2A," "PD-1," "PD-L1," "EGFR," and "HER2" as used herein refer to the human forms of their targets. A human NKG2A polypeptide sequence is available under UniProt Accession No. P26715 (SEQ ID NO: 171). A human PD-1 polypeptide sequence is available under UniProt Accession No. Q15116 (SEQ ID NO: 172). A human PD-L1 polypeptide sequence is available under UniProt Accession No. Q9NZQ7 (SEQ ID NO: 173). The human EGFR polypeptide sequence is available under UniProt accession number P00533 (SEQ ID NO: 174). The human HER2 polypeptide sequence is available under UniProt accession number P04626 (SEQ ID NO: 175). These sequences are shown in Table 4.

[0062] The term "antibody" (Ab) or "immunoglobulin" (Ig), as used herein, refers to a tetramer comprising two heavy (H) chains (about 50-70 kDa) and two light (L) chains (about 25 kDa) interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable domain (VH) and a heavy chain constant region (CH). Each light chain is composed of a light chain variable domain (VL) and a light chain constant region (CL). The VH and VL domains can be further subdivided into regions of hypervariability called "complementarity determining regions" (CDRs), interspersed with more conserved regions called "framework regions" (FRs). Each VH and VL is composed of three CDRs (H-CDR, as used herein, designates the CDRs from the heavy chain; and L-CDR, as used herein, designates the CDRs from the light chain) and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The assignment of amino acid numbers in the heavy or light chain, and of FR and CDR regions, may be according to the IMGT® definitions (Lefranc et al., Dev Comp Immunol (2003) 27(1):55-77); EU numbering; or Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD (1987 and 1991)); Chothia & Lesk, J. Mol. Biol. (1987) 196:901-17; Chothia et al., Nature (1989) 342:878-83; MacCallum et al., J. Mol. Biol. (1996) 262:732-45; or Honegger and Pluckthun, J. Mol. Biol. (2001) 309(3):657-70.

[0063] The term "recombinant antibody" refers to an antibody expressed from a cell or cell line that contains a nucleotide sequence encoding the antibody, where the nucleotide sequence is not naturally associated with the cell.

[0064] The term "isolated protein," "isolated polypeptide," or "isolated antibody" refers to a protein, polypeptide, or antibody that, by reason of its source of origin or derivation (1) is not associated with naturally associated components which accompany it in its natural state, (2) is free of other proteins from the same species, (3) is expressed from cells from a different species, and / or (4) does not occur in nature. Thus, a polypeptide that is chemically synthesized or synthesized in a cellular system other than the cell from which it is naturally derived is "isolated" from its naturally associated components. A protein can also be rendered substantially free of naturally associated components by isolation, using protein purification techniques well known in the art.

[0065] The term "affinity" refers to a measure of the attractive force between an antigen and an antibody. The intrinsic attractive force of an antibody for an antigen is typically measured by the binding affinity equilibrium constant (K D ) for binding. D is said to specifically bind an antigen when K is ≦1 μM, e.g., ≦100 nM or ≦10 nM. D Binding affinity constants can be measured by surface plasmon resonance (BIAcore™) using, for example, an IBIS MX96 SPR system from IBIS Technologies or a Carterra LSA SPR platform, or by biolayer interferometry using, for example, an Octet™ system from ForteBio.

[0066] The term "epitope" as used herein refers to a portion (determinant) of an antigen that specifically binds to an antibody or related molecule, such as a bispecific binding molecule. Epitope determinants generally consist of chemically active surface groups of molecules, such as amino acids or carbohydrate or sugar side chains, and generally have specific three-dimensional structural characteristics, as well as specific charge characteristics. Epitopes can be "linear" or "conformational". In a linear epitope, all of the points of interaction between a protein (such as an antigen) and an interacting molecule (such as an antibody) occur linearly along the primary amino acid sequence of the protein. In a conformational epitope, the points of interaction occur across amino acid residues on the protein that are separated from each other in the primary amino acid sequence. Once a desired epitope on an antigen has been determined, it is possible to generate antibodies against that epitope using techniques well known in the art. For example, antibodies against a linear epitope can be generated, for example, by immunizing animals with a peptide having amino acid residues of the linear epitope. Antibodies to a conformational epitope can be generated, for example, by immunizing an animal with a minidomain that contains the relevant amino acid residues of the conformational epitope. Antibodies to a particular epitope can also be generated, for example, by immunizing an animal with the target molecule of interest or a relevant portion thereof, and then screening for binding to the epitope.

[0067] Whether an antibody binds to the same epitope as an antibody described herein or competes for binding with an antibody can be determined by using methods known in the art, including but not limited to competitive assays, epitope binning, and alanine scanning. In some embodiments, an antibody of the present disclosure is allowed to bind to its target under saturating conditions, and then the ability of the test antibody to bind to the target can be measured. If the test antibody can bind to the target at the same time as the reference antibody, then the test antibody binds to a different epitope than the reference antibody. However, if the test antibody cannot bind to the target at the same time, then the test antibody binds to the same epitope, an overlapping epitope, or an epitope adjacent to the epitope bound by the antibody of the present disclosure. This experiment can be performed, for example, using ELISA, RIA, BIACORE™, SPR, biolayer interferometry, or flow cytometry. To test whether an antibody cross-competes with another antibody, the competitive method described above may be used in two directions, i.e., to determine whether a known antibody blocks the test antibody, or vice versa. Such reciprocal competition experiments may be performed, for example, using an IBIS MX96 or Carterra LSA SPR instrument or an Octet™ system.

[0068] The term "antigen-binding portion" (or simply "antibody portion") of an antibody as used herein refers to one or more portions or fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that certain fragments of a full-length antibody can perform the antigen-binding function of an antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" are: (i) Fab fragment: a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) F(ab')2 fragment: a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) Fd fragment consisting of the VH and CH1 domains; (iv) Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) dAb fragment consisting of the VH domain; and (vi) an isolated complementarity determining region (CDR) capable of specifically binding to an antigen. Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked using recombinant methods by a synthetic linker that allows them to be made as a single protein chain in which the VL and VH domains pair to form a monovalent molecule (known as a single chain Fv (scFv)). Also within the present disclosure are antigen-binding molecules that include VH and / or VL. In the case of VH, the molecule may also include one or more of the CH1, hinge, CH2, or CH3 regions. Such single chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. Other forms of single chain antibodies, such as diabodies, are also encompassed. Diabodies are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow pairing between the two domains on the same chain, thereby forcing the domains to pair with complementary domains on another chain and creating two antigen-binding sites.

[0069] Antibody portions, such as Fab and F(ab')2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion of whole antibodies. Furthermore, antibodies, antibody portions, and immunoadhesin molecules can be obtained using standard recombinant DNA techniques, for example, as described herein.

[0070] The class (isotype) and subclass of an antibody can be determined by any method known in the art. Generally, the class and subclass of an antibody can be determined using an antibody specific for a particular class and subclass of antibody. Such antibodies are commercially available. The class and subclass can be determined by ELISA or Western blot and other techniques. Alternatively, the class and subclass can be determined by sequencing all or part of the constant region of the heavy and / or light chain of the antibody, comparing those amino acid sequences with known amino acid sequences of various classes and subclasses of immunoglobulins, and determining the class and subclass of the antibody.

[0071] Unless otherwise indicated, all antibody amino acid residue numbers referred to in this disclosure are residue numbers under the IMGT® numbering scheme.

[0072] Anti-NKG2A antibody In some embodiments, a treatment (e.g., monotherapy or combination therapy) or composition described herein comprises an anti-NKG2A antibody or an antigen-binding portion thereof. In certain embodiments, the anti-NKG2A antibody is an antibody referred to herein as antibody mAb1 or mAb2, or a variant of either of these, where the variant comprises, for example, certain minimal amino acid changes (e.g., potentially 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes in the framework regions) compared to the antibody without losing the antigen-binding specificity of the antibody. mAb1 comprises the heavy and light chain amino acid sequences of SEQ ID NOs: 9 and 10, respectively. mAb2 comprises the heavy and light chain amino acid sequences of SEQ ID NOs: 19 and 20, respectively.

[0073] In some embodiments, the anti-NKG2A antibody binds to the same epitope on human NKG2A that competes or cross-competes with, or is identical to, antibody mAb1 or mAb2 for binding to human NKG2A.

[0074] In some embodiments, the anti-NKG2A antibody comprises H-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 1-3 or 11-13, respectively.

[0075] In some embodiments, the anti-NKG2A antibody has a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 7 or 17. In certain embodiments, the anti-NKG2A antibody has a VH that comprises the amino acid sequence of SEQ ID NO: 7 or 17.

[0076] In some embodiments, the anti-NKG2A antibody has a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 9 or 19. In certain embodiments, the anti-NKG2A antibody has a HC that comprises the amino acid sequence of SEQ ID NO: 9 or 19.

[0077] In some embodiments, the anti-NKG2A antibody comprises L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 4-6 or 14-16, respectively.

[0078] In some embodiments, the anti-NKG2A antibody has a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 8 or 18. In certain embodiments, the anti-NKG2A antibody has a VL comprising the amino acid sequence of SEQ ID NO: 8 or 18.

[0079] In some embodiments, the anti-NKG2A antibody has a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 10 or 20. In certain embodiments, the anti-NKG2A antibody has a LC that comprises the amino acid sequence of SEQ ID NO: 10 or 20.

[0080] In some embodiments, the anti-NKG2A antibody comprises any of the above heavy chain sequences and any of the above light chain sequences.

[0081] In some embodiments, the anti-NKG2A antibody comprises the following H-CDR1-3 and L-CDR1-3 amino acid sequences: a) SEQ ID NOs: 1 to 6, respectively; or b) SEQ ID NOs: 11 to 16, respectively.

[0082] In some embodiments, the anti-NKG2A antibody comprises a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 7 and 8, respectively; or b) SEQ ID NOs: 17 and 18, respectively.

[0083] In some embodiments, the anti-NKG2A antibody comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 7 and 8, respectively; or b) SEQ ID NOs: 17 and 18, respectively.

[0084] In some embodiments, the anti-NKG2A antibody comprises a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 9 and 10, respectively; or b) SEQ ID NOs: 19 and 20, respectively.

[0085] In some embodiments, the anti-NKG2A antibody comprises a HC and a LC that comprise the following amino acid sequences: a) SEQ ID NOs: 9 and 10, respectively; or b) SEQ ID NOs: 19 and 20, respectively.

[0086] In some embodiments, the anti-NKG2A antibody or antigen-binding portion thereof described herein is an anti-NKG2A antibody or antigen-binding portion thereof described in U.S. Provisional Patent Application No. 63 / 195,470, which is incorporated by reference in its entirety.

[0087] Anti-PD-1 antibody In some embodiments, the combination therapy or composition described herein comprises an anti-PD-1 antibody or antigen-binding portion thereof. In certain embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3 (comprising the heavy and light chain amino acid sequences of SEQ ID NOs: 69 and 70, respectively), retifanlimab, or a variant of any of these, where the variant can include certain minimal amino acid changes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which can be, for example, in the framework regions) compared to the antibody without losing the antigen-binding specificity of the antibody.

[0088] In some embodiments, the anti-PD-1 antibody competes or cross-competes with, or binds to the same epitope on human PD-1 as, nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab for binding to human PD-1.

[0089] In some embodiments, the anti-PD-1 antibody comprises the H-CDR1-3 and L-CDR1-3 of nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab.

[0090] In some embodiments, the anti-PD-1 antibody comprises a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the VH and VL of nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab.

[0091] In some embodiments, the anti-PD-1 antibody comprises the VH and VL of nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab.

[0092] In some embodiments, the anti-PD-1 antibody comprises a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the HC and LC of nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab.

[0093] In some embodiments, the anti-PD-1 antibody comprises the HC and LC of nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab.

[0094] In some embodiments, the anti-PD-1 antibody comprises H-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 21-23, 31-33, 41-43, 51-53, 61-63, or 71-73, respectively.

[0095] In some embodiments, the anti-PD-1 antibody has a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 27, 37, 47, 57, 67, or 77. In certain embodiments, the anti-PD-1 antibody has a VH comprising the amino acid sequence of SEQ ID NO: 27, 37, 47, 57, 67, or 77.

[0096] In some embodiments, the anti-PD-1 antibody has a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 29, 39, 49, 59, 69, or 79. In certain embodiments, the anti-PD-1 antibody has a HC that comprises the amino acid sequence of SEQ ID NO: 29, 39, 49, 59, 69, or 79.

[0097] In some embodiments, the anti-PD-1 antibody comprises L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 24-26, 34-36, 44-46, 54-56, 64-66, or 74-76, respectively.

[0098] In some embodiments, the anti-PD-1 antibody has a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 28, 38, 48, 58, 68, or 78. In certain embodiments, the anti-PD-1 antibody has a VL comprising the amino acid sequence of SEQ ID NO: 28, 38, 48, 58, 68, or 78.

[0099] In some embodiments, the anti-PD-1 antibody has a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 30, 40, 50, 60, 70, or 80. In certain embodiments, the anti-PD-1 antibody has a LC that comprises the amino acid sequence of SEQ ID NO: 30, 40, 50, 60, 70, or 80.

[0100] In some embodiments, the anti-PD-1 antibody comprises any of the above heavy chain sequences and any of the above light chain sequences.

[0101] In some embodiments, the anti-PD-1 antibody comprises the following H-CDR1-3 and L-CDR1-3 amino acid sequences: a) SEQ ID NOs: 21 to 26, respectively; b) SEQ ID NOs: 31 to 36, respectively; c) SEQ ID NOs: 41 to 46, respectively; d) SEQ ID NOs: 51 to 56, respectively; e) SEQ ID NOs: 61 to 66, respectively; or f) SEQ ID NOs: 71 to 76, respectively.

[0102] In some embodiments, the anti-PD-1 antibody comprises a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequences: a) SEQ ID NOs: 27 and 28, respectively; b) SEQ ID NOs: 37 and 38, respectively; c) SEQ ID NOs: 47 and 48, respectively; d) SEQ ID NOs: 57 and 58, respectively; e) SEQ ID NOs: 67 and 68, respectively; or f) SEQ ID NOs: 77 and 78, respectively.

[0103] In some embodiments, the anti-PD-1 antibody comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 27 and 28, respectively; b) SEQ ID NOs: 37 and 38, respectively; c) SEQ ID NOs: 47 and 48, respectively; d) SEQ ID NOs: 57 and 58, respectively; e) SEQ ID NOs: 67 and 68, respectively; or f) SEQ ID NOs: 77 and 78, respectively.

[0104] In some embodiments, the anti-PD-1 antibody comprises a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 29 and 30, respectively; b) SEQ ID NOs: 39 and 40, respectively; c) SEQ ID NOs: 49 and 50, respectively; d) SEQ ID NOs: 59 and 60, respectively; e) SEQ ID NOs: 69 and 70, respectively; or f) SEQ ID NOs: 79 and 80, respectively.

[0105] In some embodiments, the anti-PD-1 antibody comprises a HC and a LC that comprise the following amino acid sequences: a) SEQ ID NOs: 29 and 30, respectively; b) SEQ ID NOs: 39 and 40, respectively; c) SEQ ID NOs: 49 and 50, respectively; d) SEQ ID NOs: 59 and 60, respectively; e) SEQ ID NOs: 69 and 70, respectively; or f) SEQ ID NOs: 79 and 80, respectively.

[0106] In some embodiments, the anti-PD-1 antibodies, or antigen-binding portions thereof, described herein are the anti-PD-1 antibodies, or antigen-binding portions thereof, described in WO 2017 / 055547, which is incorporated by reference in its entirety.

[0107] Anti-PD-L1 antibody In some embodiments, the combination therapy or composition described herein comprises an anti-PD-L1 antibody, or an antigen-binding portion thereof. In certain embodiments, the anti-PD-L1 antibody is atezolizumab, avelumab, durvalumab, or a variant of any of these, where the variant comprises certain minimal amino acid changes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which can be, for example, in a framework region) compared to the antibody without losing the antigen-binding specificity of the antibody.

[0108] In some embodiments, the anti-PD-L1 antibody binds to the same epitope on human PD-L1 that competes or cross-competes with, or is associated with, atezolizumab, avelumab, or durvalumab for binding to human PD-L1.

[0109] In some embodiments, the anti-PD-L1 antibody comprises the H-CDRs 1-3 and L-CDRs 1-3 of atezolizumab, avelumab, or durvalumab.

[0110] In some embodiments, the anti-PD-L1 antibody comprises a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the VH and VL of atezolizumab, avelumab, or durvalumab.

[0111] In some embodiments, the anti-PD-L1 antibody comprises the VH and VL of atezolizumab, avelumab, or durvalumab.

[0112] In some embodiments, the anti-PD-L1 antibody comprises a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the HC and LC of atezolizumab, avelumab, or durvalumab.

[0113] In some embodiments, the anti-PD-L1 antibodies include the HC and LC of atezolizumab, avelumab, or durvalumab.

[0114] In some embodiments, the anti-PD-L1 antibody comprises H-CDR1-3 which comprise the amino acid sequences of SEQ ID NOs: 81-83, 91-93, or 101-103, respectively.

[0115] In some embodiments, the anti-PD-L1 antibody has a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 87, 97, or 107. In certain embodiments, the anti-PD-L1 antibody has a VH comprising the amino acid sequence of SEQ ID NO: 87, 97, or 107.

[0116] In some embodiments, the anti-PD-L1 antibody has a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 89, 99, or 109. In certain embodiments, the anti-PD-L1 antibody has a HC comprising the amino acid sequence of SEQ ID NO: 89, 99, or 109.

[0117] In some embodiments, the anti-PD-L1 antibody comprises L-CDR1-3 that comprise the amino acid sequences of SEQ ID NOs: 84-86, 94-96, or 104-106, respectively.

[0118] In some embodiments, the anti-PD-L1 antibody has a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) of the amino acid sequence of SEQ ID NO: 88, 98, or 108. In certain embodiments, the anti-PD-L1 antibody has a VL that comprises the amino acid sequence of SEQ ID NO: 88, 98, or 108.

[0119] In some embodiments, the anti-PD-L1 antibody has a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 90, 100, or 110. In certain embodiments, the anti-PD-L1 antibody has a LC comprising the amino acid sequence of SEQ ID NO: 90, 100, or 110.

[0120] In some embodiments, the anti-PD-L1 antibody comprises any of the above heavy chain sequences and any of the above light chain sequences.

[0121] In some embodiments, the anti-PD-L1 antibody comprises the following H-CDR1-3 and L-CDR1-3 amino acid sequences: a) SEQ ID NOs: 81 to 86, respectively; b) SEQ ID NOs: 91 to 96, respectively; or c) SEQ ID NOs: 101 to 106, respectively.

[0122] In some embodiments, the anti-PD-L1 antibody comprises a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequences: a) SEQ ID NOs: 87 and 88, respectively; b) SEQ ID NOs: 97 and 98, respectively; or c) SEQ ID NOs: 107 and 108, respectively.

[0123] In some embodiments, the anti-PD-L1 antibody comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 87 and 88, respectively; b) SEQ ID NOs: 97 and 98, respectively; or c) SEQ ID NOs: 107 and 108, respectively.

[0124] In some embodiments, the anti-PD-L1 antibody comprises a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 89 and 90, respectively; b) SEQ ID NOs: 99 and 100, respectively; or c) SEQ ID NOs: 109 and 110, respectively.

[0125] In some embodiments, the anti-PD-L1 antibody comprises a HC and a LC that comprise the following amino acid sequences: a) SEQ ID NOs: 89 and 90, respectively; b) SEQ ID NOs: 99 and 100, respectively; or c) SEQ ID NOs: 109 and 110, respectively.

[0126] Anti-EGFR antibody In some embodiments, the combination therapy or composition described herein comprises an anti-EGFR antibody or an antigen-binding portion thereof. In certain embodiments, the anti-EGFR antibody is cetuximab, panitumumab, futuximab, modutuximab, or a variant of any of these, where the variant comprises certain minimal amino acid changes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which may be, for example, in the framework regions) compared to the antibody, for example, without losing the antigen-binding specificity of the antibody.

[0127] In some embodiments, the anti-EGFR antibody competes or cross-competes with, or binds to the same epitope on human EGFR as cetuximab, panitumumab, futuximab, or modotuximab for binding to human EGFR.

[0128] In some embodiments, the anti-EGFR antibody comprises the H-CDR1-3 and L-CDR1-3 of cetuximab, panitumumab, futuximab, or modotuximab.

[0129] In some embodiments, the anti-EGFR antibody comprises a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the VH and VL of cetuximab, panitumumab, futuximab, or modotuximab.

[0130] In some embodiments, the anti-EGFR antibody comprises the VH and VL of cetuximab, panitumumab, futuximab, or modotuximab.

[0131] In some embodiments, the anti-EGFR antibody comprises a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the HC and LC of cetuximab, panitumumab, futuximab, or modotuximab.

[0132] In some embodiments, the anti-EGFR antibody comprises the HC and LC of cetuximab, panitumumab, futuximab, or modotuximab.

[0133] In some embodiments, the anti-EGFR antibody comprises H-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 111-113, 121-123, 131-133, or 141-143, respectively.

[0134] In some embodiments, the anti-EGFR antibody has a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 117, 127, 137, or 147. In certain embodiments, the anti-EGFR antibody has a VH comprising the amino acid sequence of SEQ ID NO: 117, 127, 137, or 147.

[0135] In some embodiments, the anti-EGFR antibody has a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 119, 129, 139, or 149. In certain embodiments, the anti-EGFR antibody has a HC that comprises the amino acid sequence of SEQ ID NO: 119, 129, 139, or 149.

[0136] In some embodiments, the anti-EGFR antibody comprises L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 114-116, 124-126, 134-136, or 144-146, respectively.

[0137] In some embodiments, the anti-EGFR antibody has a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 118, 128, 138, or 148. In certain embodiments, the anti-EGFR antibody has a VL comprising the amino acid sequence of SEQ ID NO: 118, 128, 138, or 148.

[0138] In some embodiments, the anti-EGFR antibody has a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 120, 130, 140, or 150. In certain embodiments, the anti-EGFR antibody has a LC that comprises the amino acid sequence of SEQ ID NO: 120, 130, 140, or 150.

[0139] In some embodiments, the anti-EGFR antibody comprises any of the above heavy chain sequences and any of the above light chain sequences.

[0140] In some embodiments, the anti-EGFR antibody comprises the following H-CDR1-3 and L-CDR1-3 amino acid sequences: a) SEQ ID NOs: 111 to 116, respectively; b) SEQ ID NOs: 121 to 126, respectively; c) SEQ ID NOs: 131 to 136, respectively; or d) SEQ ID NOs: 141 to 146, respectively.

[0141] In some embodiments, the anti-EGFR antibody comprises a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequences: a) SEQ ID NOs: 117 and 118, respectively; b) SEQ ID NOs: 127 and 128, respectively; c) SEQ ID NOs: 137 and 138, respectively; or d) SEQ ID NOs: 147 and 148, respectively.

[0142] In some embodiments, the anti-EGFR antibody comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 117 and 118, respectively; b) SEQ ID NOs: 127 and 128, respectively; c) SEQ ID NOs: 137 and 138, respectively; or d) SEQ ID NOs: 147 and 148, respectively.

[0143] In some embodiments, the anti-EGFR antibody comprises a HC and a LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 119 and 120, respectively; b) SEQ ID NOs: 129 and 130, respectively; c) SEQ ID NOs: 139 and 140, respectively; or d) SEQ ID NOs: 149 and 150, respectively.

[0144] In some embodiments, the anti-EGFR antibody comprises a HC and a LC that comprise the following amino acid sequences: a) SEQ ID NOs: 119 and 120, respectively; b) SEQ ID NOs: 129 and 130, respectively; c) SEQ ID NOs: 139 and 140, respectively; or d) SEQ ID NOs: 149 and 150, respectively.

[0145] In some embodiments, an anti-EGFR combination of futuximab and modotuximab (e.g., in a 1:1 ratio), or a combination including a variant of one or both antibodies, may be used where a combination therapy or composition of the disclosure calls for an "anti-EGFR antibody." In some embodiments, the anti-EGFR antibody or anti-EGFR combination is referred to as an "anti-EGFR component."

[0146] In some embodiments, the anti-EGFR combination comprises a first and a second antibody that competes or cross-competes with, or binds to the same epitope on human EGFR as, futuximab and modotuximab, respectively, for binding to human EGFR.

[0147] In some embodiments, the anti-EGFR combination comprises a first and a second antibody that comprise the H-CDR1-3 and L-CDR1-3 of futuximab and modotuximab, respectively.

[0148] In some embodiments, the anti-EGFR combination includes a first and a second antibody that comprise a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the VH and VL of futuximab and modotuximab, respectively.

[0149] In some embodiments, the anti-EGFR combination comprises a first and a second antibody comprising the VH and VL of futuximab and modotuximab, respectively.

[0150] In some embodiments, the anti-EGFR combination includes first and second antibodies that comprise a HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the HC and LC of futuximab and modotuximab, respectively.

[0151] In some embodiments, the anti-EGFR combination comprises a first and a second antibody that comprises the HC and LC of futuximab and modotuximab, respectively.

[0152] In some embodiments, the anti-EGFR combination comprises a first and a second antibody having H-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 131-133 and 141-143, respectively.

[0153] In some embodiments, the anti-EGFR combination comprises a first and a second antibody each comprising a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 137 and the amino acid sequence of SEQ ID NO: 147. In certain embodiments, the anti-EGFR combination comprises a first antibody having a VH that comprises the amino acid sequence of SEQ ID NO: 137, and a second antibody having a VH that comprises the amino acid sequence of SEQ ID NO: 147.

[0154] In some embodiments, the anti-EGFR combination comprises a first and a second antibody that each comprise a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 139 and the amino acid sequence of SEQ ID NO: 149. In certain embodiments, the anti-EGFR combination comprises a first antibody having a HC that comprises the amino acid sequence of SEQ ID NO: 139, and a second antibody having a HC that comprises the amino acid sequence of SEQ ID NO: 149.

[0155] In some embodiments, the anti-EGFR combination comprises a first and a second antibody having L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 134-136 and 144-146, respectively.

[0156] In some embodiments, the anti-EGFR combination comprises a first and a second antibody comprising a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 138 and the amino acid sequence of SEQ ID NO: 148, respectively. In certain embodiments, the anti-EGFR combination comprises a first antibody having a VL comprising the amino acid sequence of SEQ ID NO: 138, and a second antibody having a VL comprising the amino acid sequence of SEQ ID NO: 148.

[0157] In some embodiments, the anti-EGFR combination comprises a first and a second antibody that respectively comprise a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 140 and the amino acid sequence of SEQ ID NO: 150. In certain embodiments, the anti-EGFR combination comprises a first antibody having a LC that comprises the amino acid sequence of SEQ ID NO: 140 and a second antibody having a LC that comprises the amino acid sequence of SEQ ID NO: 150.

[0158] In some embodiments, the anti-EGFR combination comprises a first and a second antibody having any of the above heavy chain sequences and any of the above light chain sequences for the first and second antibodies, respectively.

[0159] In some embodiments, the anti-EGFR combination includes: a) a first antibody comprising the H-CDR1 to 3 and L-CDR1 to 3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively; and b) A second antibody comprising the amino acid sequences of H-CDR1 to 3 and L-CDR1 to 3 of SEQ ID NOs: 141 to 146, respectively.

[0160] In some embodiments, the anti-EGFR combination includes: a) a first antibody comprising a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequences of SEQ ID NOs: 137 and 138, respectively; and b) a second antibody comprising a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequences of SEQ ID NOs: 147 and 148, respectively.

[0161] In some embodiments, the anti-EGFR combination includes: a) a first antibody comprising a VH and a VL comprising the amino acid sequences of SEQ ID NOs: 137 and 138, respectively; and b) A second antibody comprising a VH and a VL comprising the amino acid sequences of SEQ ID NOs: 147 and 148, respectively.

[0162] In some embodiments, the anti-EGFR combination includes: a) a first antibody comprising a HC and a LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequences of SEQ ID NOs: 139 and 140, respectively; and b) a second antibody comprising an HC and an LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0163] In some embodiments, the anti-EGFR combination includes: a) a first antibody comprising a HC and a LC comprising the amino acid sequences of SEQ ID NOs: 139 and 140, respectively; and b) A second antibody comprising a HC and a LC comprising the amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0164]

[0001] In some embodiments, the anti-EGFR antibodies or antigen-binding portions thereof or anti-EGFR combinations described herein are anti-EGFR antibodies or antigen-binding portions thereof or combinations described in International Publication No. WO 2008 / 104183, which is incorporated by reference in its entirety into this specification.

[0165] Anti-HER2 antibody In some embodiments, the combination therapy or composition described herein comprises an anti-HER2 antibody or an antigen-binding portion thereof. In certain embodiments, the anti-HER2 antibody is trastuzumab or margetuximab, or a variant of either of these, where the variant may include certain minimal amino acid changes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which may be, for example, in the framework regions) compared to the antibody without losing the antigen-binding specificity of the antibody.

[0166] In some embodiments, the anti-HER2 antibody binds to the same epitope on human HER2 that competes or cross-competes with, or is identical to, trastuzumab or margetuximab for binding to human HER2.

[0167] In some embodiments, the anti-HER2 antibody comprises H-CDR1-3 and L-CDR1-3 of trastuzumab or margetuximab.

[0168] In some embodiments, the anti-HER2 antibody comprises a VH and VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the VH and VL of trastuzumab or margetuximab.

[0169] In some embodiments, the anti-HER2 antibody comprises the VH and VL of trastuzumab or margetuximab.

[0170] In some embodiments, the anti-HER2 antibody comprises an HC and LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the HC and LC of trastuzumab or margetuximab.

[0171] In some embodiments, the anti-HER2 antibody comprises the HC and LC of trastuzumab or margetuximab.

[0172] In some embodiments, the anti-HER2 antibody comprises H-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 151-153 or 161-163, respectively.

[0173] In some embodiments, the anti-HER2 antibody has a VH that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 157 or 167. In certain embodiments, the anti-HER2 antibody has a VH that comprises the amino acid sequence of SEQ ID NO: 157 or 167.

[0174] In some embodiments, the anti-HER2 antibody has a HC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 159 or 169. In certain embodiments, the anti-HER2 antibody has a HC that comprises the amino acid sequence of SEQ ID NO: 159 or 169.

[0175] In some embodiments, the anti-HER2 antibody comprises L-CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 154-156 or 164-166, respectively.

[0176] In some embodiments, the anti-HER2 antibody has a VL that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 158 or 168. In certain embodiments, the anti-HER2 antibody has a VL comprising the amino acid sequence of SEQ ID NO: 158 or 168.

[0177] In some embodiments, the anti-HER2 antibody has a LC that is at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 160 or 170. In certain embodiments, the anti-HER2 antibody has a LC that comprises the amino acid sequence of SEQ ID NO: 160 or 170.

[0178] In some embodiments, the anti-HER2 antibody comprises any of the above heavy chain sequences and any of the above light chain sequences.

[0179] In some embodiments, the anti-HER2 antibody comprises the following H-CDR1-3 and L-CDR1-3 amino acid sequences: a) SEQ ID NOs: 151 to 156, respectively; or b) SEQ ID NOs: 161 to 166, respectively.

[0180] In some embodiments, the anti-HER2 antibody comprises a VH and a VL that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the following amino acid sequence: a) SEQ ID NOs: 157 and 158, respectively; or b) SEQ ID NOs: 167 and 168, respectively.

[0181] In some embodiments, the anti-HER2 antibody comprises a VH and a VL comprising the following amino acid sequences: a) SEQ ID NOs: 157 and 158, respectively; or b) SEQ ID NOs: 167 and 168, respectively.

[0182] In some embodiments, the anti-HER2 antibody comprises a HC and a LC that are at least 90% (e.g., at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to: a) SEQ ID NOs: 159 and 160, respectively; or b) SEQ ID NOs: 169 and 170, respectively.

[0183] In some embodiments, the anti-HER2 antibody comprises a HC and a LC that comprise the following amino acid sequences: a) SEQ ID NOs: 159 and 160, respectively; or b) SEQ ID NOs: 169 and 170, respectively.

[0184] In some embodiments, an anti-HER2 antibody drug conjugate (ADC) may be used when a combination therapy or composition of the disclosure requires an "anti-HER2 antibody." In certain embodiments, the ADC comprises an anti-HER2 antibody described herein and DXd or DM1. In certain embodiments, the ADC is trastuzumab dexrutecan or trastuzumab emtansine.

[0185] The class of the antibodies described herein can be changed or switched to another class or subclass. In some embodiments of the present disclosure, a nucleic acid molecule encoding a VL or VH is isolated using methods well known in the art such that it does not contain a nucleic acid sequence encoding a CL or CH, respectively. The nucleic acid molecule encoding a VL or VH is then operably linked to a nucleic acid sequence encoding a CL or CH, respectively, from an immunoglobulin molecule of a different class. This can be accomplished using a vector or nucleic acid molecule containing a CL or CH sequence, as described above. For example, an antibody that was originally IgM can be class switched to an IgG. Furthermore, class switching can be used to convert one IgG subclass to another subclass, for example, from IgG1 to IgG2. The κ light chain constant region can be changed, for example, to a λ light chain constant region, or vice versa. An exemplary method for producing an antibody described herein having a desired Ig isotype includes isolating a nucleic acid molecule encoding the heavy chain of the antibody and a nucleic acid molecule encoding the light chain of the antibody, obtaining the variable domain of the heavy chain, joining the coding sequence for the variable domain of the heavy chain to a coding sequence for a constant region of the heavy chain of the desired isotype, expressing the light and heavy chains encoded by the joined sequences in a cell, and collecting the antibody having the desired isotype.

[0186] The antibodies described herein may be IgG, IgM, IgE, IgA, or IgD molecules, but will typically be of the IgG isotype, e.g., IgG subclasses IgG1, IgG 2a or IgG 2b , IgG3 or IgG4. In some embodiments, the antibody is of isotype subclass IgG1.

[0187] In some embodiments, the antibody may comprise at least one mutation in the Fc region. Many different Fc mutations are known, where these mutations alter the effector function of the antibody. For example, in some embodiments, the antibody comprises at least one mutation in the Fc region that reduces effector function, such as a mutation at one or more of positions 228, 233, 234, and 235, where the amino acid positions are numbered according to the Eu numbering system.

[0188] In some embodiments, for example, where the antibody is of the IgG1 subclass, one or both of the amino acid residues at positions 234 and 235 may be mutated, for example, from Leu to Ala (L234A / L235A). These mutations reduce the effector function of the Fc region of an IgG1 antibody. The amino acid positions are numbered according to the Eu numbering scheme.

[0189] In some embodiments, for example, when the antibody is of the IgG4 subclass, it may contain the mutation S228P, where the amino acid positions are numbered according to the Eu numbering scheme, which is known to reduce undesired Fab arm exchange.

[0190] In certain embodiments, the antibodies or antigen-binding portions thereof described herein may be part of a larger immunoadhesin molecule formed by covalent or non-covalent association of the antibody or antibody portion with one or more other proteins or peptides. Examples of such immunoadhesin molecules include the use of the streptavidin core region to generate tetrameric scFv molecules (Kipriyanov et al., Human Antibodies and Hybridomas (1995) 6: 93-101) and the use of cysteine ​​residues, marker peptides and C-terminal polyhistidine tags to generate bivalent and biotinylated scFv molecules (Kipriyanov et al., Mol. Immunol. (1994) 31: 1047-58). Other examples include where one or more CDRs from an antibody are incorporated, either covalently or non-covalently, into a molecule to make it an immunoadhesin that specifically binds to the antigen of interest. In such embodiments, the CDRs may be incorporated as part of a larger polypeptide chain, may be covalently linked to another polypeptide chain, or may be incorporated non-covalently.

[0191] In another aspect, fusion antibodies or immunoadhesins can be made that include all or a portion of an antibody described herein linked to another polypeptide. In certain embodiments, only the variable domain of the antibody is linked to the polypeptide. In certain embodiments, the VH domain of the antibody is linked to a first polypeptide, while the VL domain of the antibody is linked to a second polypeptide that associates with the first polypeptide in such a manner that the VH and VL domains can interact with each other to form an antigen binding site. In some embodiments, the VH domain is separated from the VL domain by a linker (e.g., a single chain antibody) so that the VH and VL domains can interact with each other. The VH-linker-VL antibody is then linked to a polypeptide of interest. Also, fusion antibodies can be made in which two (or more) single chain antibodies are linked to each other. This is useful when one wants to create a bivalent or multivalent antibody on a single polypeptide chain, or when one wants to create a bispecific antibody.

[0192] To make a single chain antibody (scFv), the DNA fragments encoding the VH and VL can be operably linked to another fragment encoding a flexible linker, e.g., encoding the amino acid sequence (Gly4-Ser)3 (SEQ ID NO: 176), and the VH and VL sequences can be expressed as a contiguous single chain protein with the VL and VH domains connected by a flexible linker. See, e.g., Bird et al., Science (1988) 242:423 6; Huston et al., Proc. Natl. Acad. Sci. USA (1988) 85:5879 83; and McCafferty et al., Nature (1990) 348: 552-4. Single chain antibodies can be monovalent, when only a single VH and VL are used; bivalent, when two VH and VL are used; or multivalent, when more than two VH and VL are used. For example, multispecific or multivalent antibodies can be generated that specifically bind to the targets described herein.

[0193] In other embodiments, other modified antibodies may be prepared using the antibody-encoding nucleic acid molecules. For example, "kappabodies" (Ill et al., Protein Eng. (1997) 10:949-57), "minibodies" (Martin et al., EMBO J. (1994) 13:5303-9), "diabodies" (Holliger et al., Proc. Natl. Acad. Sci. USA (1993) 90:6444-8), or "Janusins" (Traunecker et al., EMBO J. (1991) 10:3655-9 and Traunecker et al., Int. J. Cancer (Suppl.) (1992) 7:51-2) may be prepared using standard molecular biology techniques following the teachings herein.

[0194] The antibodies or antigen-binding portions described herein can be derivatized or linked to another molecule (e.g., another peptide or protein). Generally, the antibodies or portions thereof are derivatized such that target binding is not adversely affected by the derivatization or labeling. Thus, the antibodies and antibody portions of the present disclosure are intended to include both intact and modified forms of the antibodies described herein. For example, the antibodies or antibody portions of the present disclosure can be functionally linked (by chemical coupling, genetic fusion, non-covalent association, or other methods) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or diabody), a detection agent, a pharmaceutical agent, and / or a protein or peptide that can mediate the association of the antibody or antibody portion with another molecule (such as, for example, a streptavidin core region or a polyhistidine tag).

[0195] One type of derivatized antibody is produced by crosslinking two or more antibodies (of the same or different types, e.g., to create bispecific antibodies). Suitable crosslinkers include crosslinkers that are heterobifunctional (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional (e.g., disuccinimidyl suberate), having two different reactive groups separated by a suitable spacer. Such linkers are available, for example, from Pierce Chemical Company (Rockford, Ill.).

[0196] An antibody or antigen-binding portion can also be derivatized with a chemical group, such as polyethylene glycol (PEG), a methyl or ethyl group, or a carbohydrate group. These groups can be useful to improve the biological characteristics of the antibody (e.g., to increase serum half-life).

[0197] The antibodies or antigen-binding portions described herein may also be labeled. As used herein, the term "label" or "labeled" refers to the incorporation of another molecule in an antibody. In some embodiments, the label is a detectable marker, such as the incorporation of a radioactively labeled amino acid or the attachment of a biotinyl moiety to the polypeptide that can be detected by labeled avidin (e.g., streptavidin with a fluorescent marker, or enzyme activity that can be detected by optical or colorimetric methods). In some embodiments, the label or marker may be therapeutic, such as a drug conjugate or a toxin. A variety of methods of labeling polypeptides and glycoproteins are known in the art and may be used. Examples of labels for polypeptides include, but are not limited to, radioisotopes or radionuclides (e.g., 3H, 14C, 15N, 35S, 90Y, 99Tc, 111In, 125I, 131I), fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent markers, biotinyl groups, predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags). ), magnetic agents such as gadolinium chelates, toxins such as pertussis toxin, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, and analogs or homologs thereof. In some embodiments, labels are attached by spacer arms of various lengths to reduce potential steric hindrance.

[0198] In some embodiments, the antibodies or antigen-binding portions described herein may be conjugated to a cytotoxic agent to form an immunoconjugate. In some embodiments, the antibodies or antigen-binding portions according to the present disclosure may be conjugated to a radioisotope.

[0199] In certain embodiments, the antibodies described herein may exist in neutral form (including zwitterionic form) or as positively or negatively charged species. In some embodiments, the antibodies may be conjugated with a counterion to form a pharma- ceutically acceptable salt.

[0200] Combination therapy In addition to monotherapy comprising an anti-NKG2A antibody, the present disclosure provides combination therapies comprising an anti-NKG2A antibody, or an antigen-binding portion thereof, in combination with (1) an anti-PD-1 antibody, (2) an anti-PD-L1 antibody, (3) an anti-EGFR antibody, (4) an anti-HER2 antibody, or (5) any combination thereof. In certain embodiments, the combination therapy includes: a) an anti-NKG2A antibody or an antigen-binding portion thereof, and b) an anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof.

[0201] In certain embodiments, the combination therapy includes: a) an anti-NKG2A antibody or an antigen-binding portion thereof, and b) An anti-EGFR antibody or an anti-HER2 antibody, or an antigen-binding portion thereof.

[0202] In certain embodiments, the combination therapy includes: a) an anti-NKG2A antibody or an antigen-binding portion thereof; b) an anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof, and c) An anti-EGFR antibody or an anti-HER2 antibody, or an antigen-binding portion thereof.

[0203] The anti-NKG2A antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-EGFR antibody, and / or anti-HER2 antibody may be an antibody against the targets described herein. The combination therapy may take the form of, for example, a method of treatment using the antibody or antigen-binding portion, or a pharmaceutical composition comprising the antibody or antigen-binding portion.

[0204] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2), and b) An anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab).

[0205] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) An anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab).

[0206] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) An anti-EGFR antibody or antigen-binding portion thereof described herein (e.g., cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab). In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) An anti-HER2 antibody or antigen-binding portion thereof described herein (e.g., trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine).

[0207] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab), and c) An anti-EGFR antibody or antigen-binding portion thereof described herein (e.g., cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab).

[0208] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab), and c) An anti-EGFR antibody or antigen-binding portion thereof described herein (e.g., cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab).

[0209] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab), and c) An anti-HER2 antibody or antigen-binding portion thereof described herein (e.g., trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine).

[0210] In some embodiments, a combination therapy or composition of the disclosure includes: a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab), and c) An anti-HER2 antibody or antigen-binding portion thereof described herein (e.g., trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine).

[0211] In some embodiments, a combination therapy or composition of the disclosure includes: (1) mAb1, and (2) mAb3; (1) mAb1, and (2) pembrolizumab; (1) mAb1, and (2) cetuximab; (1) mAb1, and (2) futuximab + modotuximab; (1) mAb1, and (2) avelumab; (1) mAb1, (2) mAb3, and (3) futuximab + modotuximab; (1) mAb1, (2) mAb3, and (3) margetuximab; (1) mAb1, (2) retifanlimab, and (3) margetuximab; (1) mAb1, (2) pembrolizumab, and (3) trastuzumab; (1) mAb1, (2) nivolumab, and (3) trastuzumab; (1) mAb1, (2) pembrolizumab, and (3) futuximab + modotuximab; (1) mAb1, (2) nivolumab, and (3) futuximab + modotuximab; (1) mAb1, (2) nivolumab, and (3) cetuximab; or (1) mAb1, (2) pembrolizumab, and (3) cetuximab.

[0212] In some embodiments, the combination therapy or composition comprises an antibody or antigen-binding portion having the six CDRs, a VH and a VL, or a HC and LC of that antibody.

[0213] The sequences for the above referenced antibodies can be, for example, those found in Table 4. SEQ ID NOs for these sequences are assigned as shown in Table 1 below. [Table 1]

[0214] In certain embodiments, the combination therapy or composition of the disclosure includes: - anti-NKG2A antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; anti-PD-1 antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 61 to 66, respectively; and anti-EGFR combinations comprising antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; - an anti-NKG2A antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 67 and 68, respectively; and an anti-EGFR combination comprising an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and an anti-EGFR combination comprising an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0215] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 61 to 66, respectively; and an anti-HER2 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 161 to 166, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 67 and 68, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 167 and 168, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 69 and 70, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0216] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 71 to 76, respectively; and an anti-HER2 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 161 to 166, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 77 and 78, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 167 and 168, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 79 and 80, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 169 and 170, respectively.

[0217] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; and an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; and an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; and an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively.

[0218] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and an anti-HER2 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 151 to 156, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 157 and 158, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 159 and 160, respectively.

[0219] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and an anti-HER2 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 151 to 156, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-HER2 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 157 and 158, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and an anti-HER2 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 159 and 160, respectively.

[0220] In certain embodiments, the combination therapy or composition of the disclosure includes: - anti-NKG2A antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; anti-PD-1 antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and anti-EGFR combinations comprising antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; - an anti-NKG2A antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-EGFR combination comprising an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; and an anti-EGFR combination comprising an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0221] In certain embodiments, the combination therapy or composition of the disclosure includes: - anti-NKG2A antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; anti-PD-1 antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and anti-EGFR combinations comprising antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 131 to 136, respectively, and antibodies or antigen-binding portions thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 141 to 146, respectively; - an anti-NKG2A antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-EGFR combination comprising an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 137 and 138, respectively, and an antibody, or antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 147 and 148, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and an anti-EGFR combination comprising an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 139 and 140, respectively, and an antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 149 and 150, respectively.

[0222] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 31 to 36, respectively; and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 111 to 116, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 37 and 38, respectively; and an anti-EGFR antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 117 and 118, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively; and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 119 and 120, respectively.

[0223] In certain embodiments, the combination therapy or composition of the disclosure includes: - an anti-NKG2A antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 1 to 6, respectively; an anti-PD-1 antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 21 to 26, respectively; and an anti-EGFR antibody or an antigen-binding portion thereof comprising the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 111 to 116, respectively; - an anti-NKG2A antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 7 and 8, respectively; an anti-PD-1 antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 27 and 28, respectively; and an anti-EGFR antibody, or an antigen-binding portion thereof, comprising the VH and VL amino acid sequences of SEQ ID NOs: 117 and 118, respectively; or - an anti-NKG2A antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 9 and 10, respectively; an anti-PD-1 antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and an anti-EGFR antibody comprising the HC and LC amino acid sequences of SEQ ID NOs: 119 and 120, respectively.

[0224] multispecific binding molecules In a further aspect, the disclosure provides multispecific binding molecules having the binding specificity of an anti-NKG2A antibody or antigen-binding portion thereof (e.g., comprising an antigen-binding portion, such as an antigen-binding domain comprising six CDRs) in combination with the binding specificity of (1) an anti-PD-1 antibody, (2) an anti-PD-L1 antibody, (3) an anti-EGFR antibody, (4) an anti-HER2 antibody, or (5) any combination thereof. In certain embodiments, the multispecific binding molecules have the binding specificity of the following (e.g., comprising an antigen-binding portion, such as an antigen-binding domain comprising six CDRs): a) an anti-NKG2A antibody or an antigen-binding portion thereof, and b) an anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof;

[0225] In certain embodiments, a polyspecific binding molecule has the following binding specificities (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or an antigen-binding portion thereof; b) an anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof; and c) An anti-EGFR or anti-HER2 antibody, or an antigen-binding portion thereof. The anti-NKG2A antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-EGFR antibody, and / or anti-HER2 antibody may be an antibody against a target as described herein.

[0226] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2), and b) An anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab).

[0227] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2), and b) An anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab).

[0228] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab), and c) An anti-EGFR antibody or antigen-binding portion thereof described herein (e.g., cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab).

[0229] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab), and c) An anti-EGFR antibody or antigen-binding portion thereof described herein (e.g., cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab).

[0230] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-1 antibody or antigen-binding portion thereof described herein (e.g., nivolumab, pembrolizumab, cemiplimab, dostallimab, mAb3, or retifanlimab), and c) An anti-HER2 antibody or antigen-binding portion thereof described herein (e.g., trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine).

[0231] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen binding portion, such as an antigen binding domain comprising six CDRs): a) an anti-NKG2A antibody or antigen-binding portion thereof described herein (e.g., mAb1 or mAb2); b) an anti-PD-L1 antibody or antigen-binding portion thereof described herein (e.g., atezolizumab, avelumab, or durvalumab), and c) An anti-HER2 antibody or antigen-binding portion thereof described herein (e.g., trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine).

[0232] In some embodiments, a polyspecific binding molecule has the following binding specificity (e.g., comprises an antigen-binding portion, such as an antigen-binding domain comprising six CDRs or a VH and a VL): (1) mAb1, and (2) mAb3; (1) mAb1 and pembrolizumab; (1) mAb1, and (2) cetuximab; (1) mAb1, and (2) futuximab + modotuximab; (1) mAb1, and (2) avelumab; (1) mAb1, (2) mAb3, and (3) futuximab + modotuximab; (1) mAb1, (2) mAb3, and (3) margetuximab; (1) mAb1, (2) retifanlimab, and (3) margetuximab; (1) mAb1, (2) pembrolizumab, and (3) trastuzumab; (1) mAb1, (2) nivolumab, and (3) trastuzumab; (1) mAb1, (2) pembrolizumab, and (3) futuximab + modotuximab; (1) mAb1, (2) nivolumab, and (3) futuximab + modotuximab; (1) mAb1, (2) nivolumab, and (3) cetuximab, or (1) mAb1, (2) pembrolizumab, and (3) cetuximab.

[0233] Polyspecific binding molecules are known in the art, and examples of different types of polyspecific binding molecules are provided elsewhere herein. Such polyspecific (e.g., bispecific or trispecific) binding molecules are encompassed by the treatments of the present disclosure.

[0234] Nucleic Acid Molecules and Vectors Also described are the nucleic acid molecules and sequences, antibodies or antigen-binding portions thereof, described herein. In some embodiments, different nucleic acid molecules encode the heavy and light chain amino acid sequences of the antibody or antigen-binding portion. In other embodiments, the same nucleic acid molecule encodes the heavy and light chain amino acid sequences of the antibody or antigen-binding portion.

[0235] A reference to a nucleotide sequence includes its complement unless otherwise specified. Thus, a reference to a nucleic acid having a particular sequence should be understood to include its complementary strand, with its complementary sequence. The term "polynucleotide" as referred to herein means a polymeric form of nucleotides of at least 10 bases in length, either ribonucleotides or deoxynucleotides, or a modified form of either type of nucleotide. The term includes single- and double-stranded forms.

[0236] In some embodiments, the disclosure provides a nucleic acid molecule comprising a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, or a nucleotide sequence encoding the light chain or antigen-binding portion thereof, of an antibody or antigen-binding portion thereof described herein, or both.

[0237] The present disclosure also provides nucleotide sequences that are at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% identical to a nucleotide sequence that encodes an amino acid sequence selected from the group consisting of SEQ ID NOs: 7-10, 17-20, 27-30, 37-40, 47-50, 57-60, 67-70, 77-80, 87-90, 97-100, 107-110, 117-120, 127-130, 137-140, 147-150, 157-160, or 167-170. The term "percent sequence identity" in the context of nucleic acid sequences refers to the residues in two sequences that are identical when aligned for maximum correspondence. The length of sequence identity comparison can be over a stretch of at least about 9 nucleotides, usually at least about 18 nucleotides, more usually at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 32 nucleotides, preferably at least about 36, 48 or more nucleotides. There are many different algorithms known in the art that can be used to measure nucleotide sequence identity. For example, polynucleotide sequences can be compared using FASTA, Gap or Bestfit, which are programs in the Wisconsin Package Version 10.0 Genetics Computer Group (GCG) (Madison, Wisconsin). FASTA, for example, includes the programs FASTA2 and FASTA3, which provide alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (see, e.g., Pearson, Methods Enzymol. (1990) 183:63-98; Pearson, Methods Mol. Biol. (2000) 132:185-219; Pearson, Methods Enzymol. (1996) 266:227-58; and Pearson, J. Mol. Biol. (1998) 276:71-84; incorporated herein by reference). Unless otherwise specified, default parameters for a particular program or algorithm are used.For example, percent sequence identity between nucleic acid sequences can be determined using FASTA with its default parameters (NOPAM coefficient for a word size and scoring matrix of 6) or Gap with its default parameters provided in GCG version 6.1, which is incorporated herein by reference.

[0238] In any of the above embodiments, the nucleic acid molecule may be isolated. Nucleic acid molecules referred to herein as "isolated" or "purified" are (1) separated from their source genomic DNA or cellular RNA nucleic acid; and / or (2) nucleic acids that do not occur in nature.

[0239] In a further aspect, the disclosure provides vectors suitable for expressing one or both chains of an antibody or antigen-binding portion thereof described herein. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. In some embodiments, a vector is a plasmid, i.e., a circular double-stranded piece of DNA to which additional DNA segments can be ligated. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors").

[0240] The present disclosure provides vectors comprising nucleic acid molecules encoding the heavy chain, light chain, or both the heavy and light chains of an antibody or antigen-binding portion thereof described herein. In certain embodiments, the vectors of the present disclosure comprise nucleic acid molecules described herein. The present disclosure further provides vectors comprising nucleic acid molecules encoding fusion proteins, modified antibodies, antibody fragments, and probes thereof. The vectors may further comprise expression control sequences.

[0241] The term "expression control sequences" as used herein refers to polynucleotide sequences necessary to affect the expression and processing of coding sequences to which they are linked. Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals, such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequences); sequences that enhance protein stability; and, if desired, sequences that enhance protein secretion. The nature of such control sequences varies depending on the host organism; in prokaryotes, such control sequences generally include promoters, ribosomal binding sites, and transcription termination sequences; in eukaryotes, such control sequences generally include promoters and transcription termination sequences. The term "control sequences" is intended to include, at a minimum, all components whose presence is essential for expression and processing, and can also include additional components whose presence is advantageous, such as leader sequences and fusion partner sequences.

[0242] In some embodiments, the nucleic acid molecules described herein comprise a nucleotide sequence encoding a VH domain from an antibody or antigen-binding portion described herein linked in-frame to a nucleotide sequence encoding a heavy chain constant region from any source. Similarly, the nucleic acid molecules described herein can comprise a nucleotide sequence encoding a VL domain from an antibody or antigen-binding portion described herein linked in-frame to a nucleotide sequence encoding a light chain constant region from any source.

[0243] In a further aspect of the disclosure, nucleic acid molecules encoding VH and / or VL can be "converted" into full-length antibody genes. In some embodiments, nucleic acid molecules encoding VH or VL domains are converted into full-length antibody genes by insertion into an expression vector already encoding a heavy chain constant (CH) region or a light chain constant (CL) region, respectively, with the VH segment operably linked to a CH segment in the vector and / or the VL segment operably linked to a CL segment in the vector. In another aspect, nucleic acid molecules encoding VH and / or VL domains are converted into full-length antibody genes by linking, e.g., ligating, the nucleic acid molecules encoding the VH and / or VL domains to nucleic acid molecules encoding the CH and / or CL regions using standard molecular biology techniques. The nucleic acid molecules encoding the full-length heavy and / or light chains can then be expressed from a cell into which they have been introduced and the antibody can be isolated.

[0244] In some embodiments, the framework regions are mutated so that the resulting framework regions have the amino acid sequence of the corresponding germline gene. Mutations are made in the framework regions or constant regions, for example, to increase the half-life of the antibody. See, for example, WO 00 / 09560. Mutations in the framework regions or constant regions can also be made to change the immunogenicity of the antibody and / or provide a site for covalent or non-covalent binding to another molecule. According to the present disclosure, the antibody can have mutations in any one or more of the CDRs or framework regions of the variable domain, or in the constant region.

[0245] Host Cells and Methods for Producing Antibodies and Antibody Compositions Also described are methods of producing the combination therapies (e.g., compositions) of the present disclosure. One embodiment is a method of producing an antibody described herein, comprising providing a recombinant host cell capable of expressing the antibody, culturing the host cell under conditions suitable for expression of the antibody, and isolating the resulting antibody. The antibody produced by such expression in such a recombinant host cell is referred to herein as a "recombinant antibody." Also described are the progeny of such host cells, and the antibodies produced thereby.

[0246] The term "recombinant host cell" (or simply "host cell"), as used herein, refers to a cell into which a recombinant expression vector has been introduced. By definition, recombinant host cells do not occur in nature. The present disclosure provides host cells that may, for example, comprise a vector described herein. The present disclosure also provides host cells that comprise a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, a nucleotide sequence encoding the light chain or antigen-binding portion thereof, or both, of an antibody or antigen-binding portion thereof described herein. It should be understood that "recombinant host cell" and "host cell" refer not only to the particular subject cell, but also to the progeny of such a cell. Because certain modifications may occur in successive generations, either due to mutation or environmental influences, such progeny will not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0247] Nucleic acid molecules encoding the antibodies or antigen-binding portions thereof described herein, and vectors containing these nucleic acid molecules, can be used for transfection of suitable mammalian, plant, bacterial or yeast host cells. Transformation can be by any known method for introducing polynucleotides into host cells. Methods for the introduction of heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of polynucleotides in liposomes, and direct microinjection of DNA into the nucleus. Nucleic acid molecules can also be introduced into mammalian cells by viral vectors.

[0248] Antibodies expressed by different cell lines or in transgenic animals are likely to have different glycosylation patterns from each other, however, all antibodies encoded by the nucleic acid molecules provided herein or comprising the amino acid sequences provided herein are part of the present disclosure, regardless of the glycosylation state of the antibody, and more generally, regardless of the presence or absence of post-translational modifications.

[0249] In some embodiments, the disclosure relates to a method for producing an antibody composition comprising an anti-NKG2A antibody, or antigen-binding portion thereof, and an anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, the method comprising: - providing first and second host cells, wherein the first host cell is capable of expressing an anti-NKG2A antibody, or an antigen-binding portion thereof, described herein, and the second host cell is capable of expressing an anti-PD-1 or PD-L1 antibody, or an antigen-binding portion thereof, described herein; - culturing the first and second antibodies and the host cell under conditions suitable for expression of the anti-NKG2A antibody, or an antigen-binding portion thereof, and the anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof; and - isolating the resulting antibody.

[0250] In some embodiments, the disclosure relates to a method for producing an antibody composition comprising an anti-NKG2A antibody, or antigen-binding portion thereof, an anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, and an anti-EGFR or anti-HER2 antibody, or antigen-binding portion thereof, the method comprising: - providing a first, second, and third host cell, wherein the first host cell is capable of expressing an anti-NKG2A antibody, or an antigen-binding portion thereof, described herein, the second host cell is capable of expressing an anti-PD-1 antibody, or an antigen-binding portion thereof, and the third host cell is capable of expressing an anti-PD-L1 antibody, or an antigen-binding portion thereof, described herein; - culturing the first, second, and third host cells under conditions suitable for expression of an anti-NKG2A antibody, or antigen-binding portion thereof, an anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, and an anti-EGFR antibody, or antigen-binding portion thereof; and - isolating the resulting antibody.

[0251] The present disclosure also provides a host cell comprising: - a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, a nucleotide sequence encoding the light chain or antigen-binding portion thereof, or both, of an anti-NKG2A antibody or antigen-binding portion thereof described herein, and a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, light chain or antigen-binding portion thereof, or both, of an anti-PD-1 or PD-L1 antibody described herein; or - a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, a nucleotide sequence encoding the light chain or antigen-binding portion thereof, or both, of an anti-NKG2A antibody or antigen-binding portion thereof described herein; a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, a nucleotide sequence encoding the light chain or antigen-binding portion thereof, or both, of an anti-PD-1 antibody or PD-L1 antibody described herein; a nucleotide sequence encoding the heavy chain or antigen-binding portion thereof, a nucleotide sequence encoding the light chain or antigen-binding portion thereof, or both, of an anti-EGFR or anti-HER2 antibody described herein.

[0252] Pharmaceutical Compositions Another aspect of the disclosure is a pharmaceutical composition that includes as an active ingredient (e.g., as the only active ingredient): - an anti-NKG2A antibody or an antigen-binding portion thereof described herein; - an anti-NKG2A antibody, or an antigen-binding portion thereof, as described herein, and an anti-PD-1 or anti-PD-L1 antibody, or an antigen-binding portion thereof, as described herein; - an anti-NKG2A antibody, or an antigen-binding portion thereof, as described herein, and an anti-EGFR or anti-HER2 antibody, or an antigen-binding portion thereof, as described herein; or - an anti-NKG2A antibody, or an antigen-binding portion thereof, as described herein, an anti-PD-1 or PD-L1 antibody, or an antigen-binding portion thereof, as described herein, and an anti-EGFR or anti-HER2 antibody, or an antigen-binding portion thereof, as described herein.

[0253] In some aspects, the pharmaceutical composition comprises a multispecific binding molecule (e.g., an anti-NKG2A antibody described herein and an anti-PD-1 or anti-PD-L1 antibody described herein; or a multispecific binding molecule having the binding specificities of an anti-NKG2A antibody, an anti-PD-1 or anti-PD-L1 antibody, and an anti-EGFR or anti-HER2 antibody described herein).

[0254] Another aspect of the present disclosure is a pharmaceutical composition comprising as an active ingredient (or as the only active ingredient) a monotherapy or combination therapy of the present disclosure. The pharmaceutical composition may additionally comprise a pharma- ceutical acceptable excipient. In some embodiments, the pharmaceutical composition is intended for the amelioration, prevention, and / or treatment of cancer, such as the cancers described herein. In certain embodiments, the cancer is in a tissue, such as skin, lung, intestine, colon, ovary, brain, prostate, kidney, soft tissue, hematopoietic system, head and neck, liver, bone, bladder, breast, stomach, uterus, cervix, and pancreas.

[0255] The pharmaceutical compositions of the present disclosure will comprise one or more of the antibodies, antigen-binding portions, antibody compositions, or multispecific binding molecules described herein. In some embodiments, the compositions comprise two antibodies or antigen-binding portions thereof described herein. In some embodiments, the compositions comprise three antibodies or antigen-binding portions thereof described herein. In some embodiments, the compositions comprise a monotherapy or combination therapy described herein.

[0256] In some embodiments, the pharmaceutical compositions may include a monotherapy or combination therapy of the present disclosure and one or more additional agents selected from, for example, immune stimulants, vaccines, chemotherapeutic agents, anti-tumor agents, anti-angiogenic agents, and tyrosine kinase inhibitors.

[0257] In some embodiments, the pharmaceutical composition is intended for the amelioration, prevention, and / or treatment of a disorder, disease, or condition that improves or slows its progression by modulating NKG2A, PD-1, PD-L1, EGFR, HER2, or any combination thereof. In some embodiments, the pharmaceutical composition is intended for the amelioration, prevention, and / or treatment of cancer. In some embodiments, the pharmaceutical composition is intended for activation of the immune system.

[0258] Generally, the treatments and compositions of the disclosure will be suitable for administration in one or more formulations in combination with one or more pharma- ceutically acceptable excipients, for example, as described below.

[0259] The term "excipient" is used herein to describe any ingredient other than the compound of the present disclosure. The choice of excipient depends to a large extent on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form. As used herein, "pharmaceutical acceptable excipient" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. Some examples of pharmaceutical acceptable excipients are water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. In many cases, it will be preferable to include isotonic agents, such as sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride in the composition. Additional examples of pharmaceutical acceptable substances are wetting agents or small amounts of auxiliary substances, such as wetting agents or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the antibody.

[0260] Pharmaceutical compositions of the present disclosure and methods for their preparation will be readily apparent to those skilled in the art. Such compositions and methods for their preparation can be found, for example, in Remington's Pharmaceutical Sciences, 19th Edition (Mack Publishing Company, 1995). Pharmaceutical compositions are preferably manufactured under GMP (Good Manufacturing Practice) conditions.

[0261] The pharmaceutical composition of the present disclosure can be prepared, packaged, or sold in bulk as a single unit dose or as a plurality of single unit doses.As used herein, a "unit dose" is a discrete amount of pharmaceutical composition that contains a predetermined amount of active ingredient.The amount of active ingredient is generally equal to the dosage of active ingredient that can be administered to a subject, or a convenient fraction of such a dosage, for example, one-half or one-third of such a dosage.

[0262] A formulation of a pharmaceutical composition suitable for parenteral administration typically comprises the active ingredient in combination with a pharma- ceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and the like. Such formulations may further comprise one or more additional components, including, but not limited to, suspending agents, stabilizing agents, or dispersing agents. In some embodiments of formulations for parenteral administration, the active ingredient is provided in a dry (i.e., powder or granules) form for reconstitution with a suitable solvent (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition. Parenteral formulations also include aqueous solutions which may contain excipients such as salts, carbohydrates, buffers, etc. (preferably pH 3-9), however, for some applications, they may be more suitably formulated as sterile nonaqueous solutions or as dry forms used in combination with suitable solvents such as sterile pyrogen-free water. Exemplary parenteral administration forms include solutions or suspensions in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose. Such dosage forms can be suitably buffered if desired. Other parenterally administrable formulations which are useful include those which contain the active ingredient in microcrystalline form or in a liposomal preparation.

[0263] Therapeutic Uses of the Antibodies and Compositions of the Disclosure In some embodiments, the treatments and compositions of the present disclosure are used to enhance or activate the immune system in a patient (e.g., a mammal, such as a human) in need thereof. In certain embodiments, the patient is immunosuppressed. In certain embodiments, a physician can boost the anti-cancer activity of the patient's own immune system by administering a treatment or composition described herein. For example, a physician can boost anti-tumor activity in a patient by administering a treatment or composition of the present disclosure alone or in combination with other therapeutic agents (sequentially or simultaneously).

[0264] In certain embodiments, the disclosed therapy or composition is for use in the treatment of cancer, which may be in one or more tissues, such as the skin, lung, intestine, colon, ovary, brain, prostate, kidney, soft tissue, hematopoietic system, head and neck, liver, bone, bladder, breast, stomach, uterus, cervix, and pancreas.

[0265] In some embodiments, cancers that are treated by the therapies and compositions of the disclosure include, for example, melanoma (e.g., advanced or metastatic melanoma), basal cell carcinoma of the skin, glioblastoma, glioma, gliosarcoma, astrocytoma, meningioma, neuroblastoma, adrenocortical carcinoma, squamous cell carcinoma of the head and neck, oral cancer, salivary gland cancer, nasopharyngeal cancer, breast cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC), small cell lung cancer, and squamous cell lung cancer), esophageal cancer, gastroesophageal junction cancer, gastric cancer, gastrointestinal cancer, primary peritoneal cancer, liver cancer, hepatocellular carcinoma, biliary tract cancer, colon cancer, rectal cancer, colorectal cancer, ovarian cancer, fallopian tube cancer, bladder cancer, upper urinary tract cancer, urothelial carcinoma, renal cell carcinoma, kidney cancer, genitourinary cancer, uterine cancer, and ovarian cancer. These may include cervical cancer, prostate cancer, fibrosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, histiocytoma, pancreatic cancer, endometrial cancer, appendix cancer, advanced Merkel cell carcinoma, multiple myeloma, sarcoma, choriocarcinoma, erythroleukemia, acute lymphocytic leukemia, acute monocytic leukemia, acute promyelocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, mast cell leukemia, small lymphocytic lymphoma, Burkitt lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, monocytic lymphoma, HTLV-associated T-cell leukemia / lymphoma, mesothelioma, and solid tumors. The cancer may be, for example, at an early, intermediate, late, locally advanced, advanced, or metastatic stage, may recur, and may be refractory and / or intolerant to other therapies (e.g., one or more targets of a treatment or composition for the cancer, a checkpoint inhibitor, or other therapy directed to the standard of care), or there may be no standard of care available. In some embodiments, the cancer will not be amenable to surgical intervention, either due to medical contraindications or inability to resect the tumor.

[0266] In some embodiments, conditions treated by the therapies and compositions of the present disclosure may include, for example, gastric and colorectal cancer, hi some embodiments, the gastric or colorectal cancer is metastatic, locally advanced, or unresectable.

[0267] In some embodiments, the gastric cancer is: (1) unresectable; (2) locally advanced or metastatic; (3) HER2 +or (4) any combination (e.g., all) of (1)-(3). Additionally or alternatively, the gastric cancer patient will have been treated with first-line standard therapy (e.g., cytotoxic chemotherapy, trastuzumab, and / or pembrolizumab). In certain embodiments, the treatment or composition of the disclosure (e.g., targeting NKG2A, PD-1, and HER2, such as mAb1+mAb3+margetuximab) is used to treat locally advanced unresectable or metastatic HER2 + It may be used to treat gastric cancer, for example, where first-line standard therapies, such as cytotoxic chemotherapy, trastuzumab, and / or pembrolizumab, have failed.

[0268] In some embodiments, the treatments or compositions of the disclosure (e.g., targeting NKG2A, PD-1, and HER2, such as mAb1 + pembrolizumab + trastuzumab) are used to treat locally advanced unresectable or metastatic HER2 + It may be used to treat gastroesophageal junction (GEJ) and gastric adenocarcinoma (GA) where, for example, first-line standard therapies, such as cytotoxic chemotherapy, trastuzumab, and / or pembrolizumab, have failed.

[0269] In some embodiments, the colorectal cancer is (1) metastatic, (2) not undergoing surgical intervention due to either medical contraindications or tumor unresectability, (3) has low microsatellite instability status (e.g., MSI-H cancer) according to institutional guidelines or College of American Pathologists guidelines, (4) any combination (e.g., all) of (1)-(3). Additionally or alternatively, the patient with colorectal cancer will (i) not have a RAS (KRAS and NRAS) mutation in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4; and / or (ii) not have a BRAF V600E mutation. In certain embodiments, therapies or compositions of the present disclosure (e.g., mAb1+mAb3+futuximab+modotuximab, etc., targeting NKG2A, PD-1, and EGFR) may be used to treat metastatic colorectal cancer. In certain embodiments, mAb1 and pembrolizumab treatments or compositions may be used to treat colorectal cancer, specifically MSI-H / dMMR locally advanced unresectable or metastatic colorectal cancer (mCRC).

[0270] In some embodiments, therapies or compositions of the disclosure may be used to treat patient populations as described in Example 10.

[0271] In some embodiments, the treatments or compositions described herein may inhibit tumor growth and / or induce tumor growth regression in vivo. In some embodiments, the treatments or compositions described herein may slow or reverse metastasis in cancer patients. In some embodiments, the treatments or compositions described herein may prolong survival of cancer patients. Any combination of the above properties is also contemplated.

[0272] In some embodiments, therapies or compositions of the disclosure may be used in the treatment of immune disorders.

[0273] In some embodiments, the treatments or compositions of the disclosure may be used to treat patients who are or are at risk of being immunocompromised (e.g., due to chemotherapy or radiation therapy), hi some embodiments, the treatments or compositions may be used to expand stem cells in patients following stem cell transplantation.

[0274] In some embodiments, the therapy or composition is for use in treating viral and / or parasitic infections, for example, where the pathogen inhibits the host immune response. The pathogen can be, for example, HIV, Hepatitis (A, B, or C), Human Papillomavirus (HPV), Lymphocytic Choriomeningitis Virus (LCMV), Adenovirus, Flavivirus, Echovirus, Rhinovirus, Coxsackievirus, Coronavirus, Respiratory Syncytial Virus, Mumps Virus, Rotavirus, Measles Virus, Rubella Virus, Parvovirus, Vaccinia Virus, Human T-cell Lymphotrophic Virus (HTLV), Human Cytomegalovirus (HCMV), Dengue Virus, Molluscum Virus, Poliovirus, Rabies Virus, John Cunningham (JC) Virus, Arboviral Encephalitis Virus, Simian Immunodeficiency Virus (SIV), Influenza, Herpes, Giardia, Malaria, Leishmania, Staphylococcus aureus, or Pseudomonas aeruginosa.

[0275] "Treat", "treating" and "treatment" refer to a method of alleviating or arresting at least one of a biological disorder and / or symptoms associated therewith. As used herein, "alleviating" a disease, disorder or condition means reducing the severity and / or frequency of occurrence of the symptoms of the disease, disorder or condition. Furthermore, references herein to "treatment" include references to curative, palliative, and prophylactic treatment.

[0276] "Therapeutically effective amount" refers to the amount of the therapeutic agent being administered that relieves to some extent one or more of the symptoms of the disease being treated. A therapeutically effective amount of an anti-cancer therapeutic agent may result in, for example, delayed tumor growth, tumor shrinkage, increased survival time, elimination of cancer cells, slowed or reduced disease progression, reversal of metastasis, or other clinical endpoints desired by a medical professional. In some embodiments, a therapeutically effective amount of a treatment or composition of the present disclosure results in, for example, improved objective response rate, improved clinical benefit rate, improved duration of response, increased progression-free survival, and increased overall survival, compared to untreated patients.

[0277] In some embodiments, the treatments described herein are administered in a single composition. In other embodiments, the treatments (e.g., combination therapies) are administered in more than one composition. For example, a combination therapy including an anti-NKG2A antibody, an anti-PD-1 or anti-PD-L1 antibody, and an anti-EGFR or anti-HER2 antibody may include administration of a single composition including all three antibodies, a composition including two of the antibodies and a composition including one of the antibodies, or a separate composition for each antibody. In cases where there is more than one composition, the compositions may be administered simultaneously, sequentially, separately, or any combination thereof.

[0278] The treatments or compositions of the present disclosure may be administered without additional therapeutic treatments, i.e., as a sole treatment (monotherapy). Alternatively, the treatment or combination treatment may include at least one additional therapeutic treatment, such as another immune stimulant, an anti-cancer agent (e.g., a chemotherapeutic agent, an anti-tumor agent, an anti-angiogenic agent, or a tyrosine kinase inhibitor), or a vaccine (e.g., a tumor vaccine).

[0279] In some embodiments, the therapy or composition may be co-administered or formulated with another medicament / drug for the treatment of cancer. The additional therapeutic treatment may include, for example, an immune stimulant, a vaccine, a chemotherapeutic agent, an anti-tumor agent, an anti-angiogenic agent, a tyrosine kinase inhibitor, and / or radiation therapy. In some embodiments, the additional therapeutic treatment may include a different anti-cancer antibody.

[0280] Pharmaceutical products containing the treatments or compositions described herein and at least one other agent (e.g., a chemotherapeutic agent, an antitumor agent, or an antiangiogenic agent) can be used as a combination treatment for simultaneous, separate, or sequential administration in cancer therapy. The other agent may be any agent suitable for treating the particular cancer in question, such as an alkylating agent, such as a platinum derivative, such as cisplatin, carboplatin, and / or oxaliplatin; a plant alkoid, such as paclitaxel, docetaxel, and / or irinotecan; an antitumor antibiotic, such as doxorubicin (adriamycin), daunorubicin, epirubicin, idarubicin, mitoxantrone, dactinomycin, bleomycin, actinomycin, luteomycin, and / or mitomycin; a topoisomerase inhibitor, such as topotecan; an antimetabolite, such as fluorouracil and / or other fluoropyrimidines; FOLFOX; osimertinib; cyclophosphamide; anthracyclines; dacarbazine; gemcitabine; or any combination thereof. In some embodiments, the treatment or composition described herein re-establishes responsiveness to the other agent.

[0281] The treatment or composition of the present disclosure may also be used in combination with other anti-cancer therapies, such as vaccines, cytokines, enzyme inhibitors, immune stimulating compounds, and T cell therapy. In the case of a vaccine, it may be, for example, a protein, peptide, or DNA vaccine that contains one or more antigens relevant for the cancer being treated, or a vaccine that contains dendritic cells with antigens. Suitable cytokines include, for example, IL-2, IFN-gamma, and GM-CSF. An example of a type of enzyme inhibitor with anti-cancer activity is an indoleamine-2,3-dioxygenase (IDO) inhibitor, such as 1-methyl-D-tryptophan (1-D-MT). Also contemplated is adoptive T cell therapy, which refers to a variety of immunotherapy techniques that involve expanding or manipulating the patient's own T cells to recognize and attack the tumor.

[0282] It is also contemplated that the treatments or compositions of the present disclosure may be used in adjunctive therapy in conjunction with tyrosine kinase inhibitors, which are synthetic, primarily quinazoline-derived, small molecular weight molecules that interact with the intracellular tyrosine kinase domain of the receptor and inhibit ligand-induced receptor phosphorylation, e.g., by competing for the intracellular Mg-ATP binding site.

[0283] In some embodiments, the treatment or composition includes, but is not limited to, A2AR, A1AR, A2BR, A3AR, ADA, ALP, AXL, BTLA, B7-H3, B7-H4, CTLA-4, CD116, CD123, CD27, CD28, CD39, CD40, CD47, CD55, CD73, CD122, CD137, CD160, CGEN-15049, CHK1, CHK2, CTLA-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), EGFR, FLT3, HER2, NKG2AL, GAL9, GITR, HVEM, LAG-3, LI It may be used in combination with medicines / drugs that mediate immune system activation, including agents that modulate the expression or activity of LRB1, LY108, LAIR1, MET, NKG2A, ICOS, IDO, IL2R, IL4R, KIR, LAIR1, PAP, PD-1 / PD-L1 / PD-L2, OX40, STING, TIGIT, TIM-3, TGFR-β, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR10, TNFR2, VEGFR, VEGF, VISTA, LILRB2, CMTM6 and / or 2B4. In certain embodiments, the agent is a small molecule inhibitor. In certain embodiments, the agent is an antibody or antigen-binding fragment thereof that binds to one of the above molecules. It is also contemplated that the treatments or compositions of the disclosure may be used in combination with cytokines (e.g., IL-1, IL-2, IL-12, IL-15, or IL-21), EGFR inhibitors, VEGF inhibitors, and the like.

[0284] As used herein, the terms "co-administration," "co-administered," and "in combination with" refer to the treatments and compositions of the disclosure with one or more other therapeutic agents, and are intended to mean, and indeed refer to and include, the following: a) simultaneous administration of such a treatment / composition of the present disclosure and a therapeutic agent to a patient in need of treatment, where such components are formulated together in a single dosage form that releases the components to the patient substantially simultaneously; b) the substantially simultaneous administration of such treatments / compositions of the present disclosure and therapeutic agents to a patient in need of treatment, where such components are formulated in separate dosage forms separate from one another, which are ingested by the patient at substantially the same time, such that the components are released to the patient at substantially the same time; c) sequential administration of such treatments / compositions and therapeutic agents of the present disclosure to a patient in need of treatment, where such components are formulated in separate dosage forms separate from one another, which are ingested by the patient at successive times, with a significant time interval between each administration, such that the components are released to the patient at substantially different times; and d) Sequential administration of such treatments / compositions of the present disclosure and therapeutic agents to a patient in need of treatment when such components are formulated together in a single dosage form that releases the components in a controlled manner, where they are released simultaneously, sequentially, and / or overlappingly to the patient at the same and / or different times, and where each portion may be administered by either the same or different routes.

[0285] It is understood that the treatments and compositions of the disclosure may be used in the methods of treatment described herein, may be for use in the treatments described herein, and / or may be for use in the manufacture of a medicament for the treatments described herein.

[0286] Dosage and Route of Administration Therapies and compositions of the present disclosure may be administered in an effective amount for the treatment of the condition in question, i.e., at dosages and for periods of time necessary to achieve the desired result. The therapeutically effective amount may vary according to factors such as the particular condition being treated, the age, sex, and weight of the patient, whether the antibody is administered as the sole treatment or in combination with one or more additional anti-cancer treatments, etc.

[0287] Dosage regimens can be adjusted to provide the optimum desired response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to physically discrete units suitable as unitary dosages for the patient / subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for dosage unit forms of the present disclosure are generally dictated by and directly dependent on (a) the unique characteristics of the therapeutic agent and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of compounding such active compounds for the treatment of susceptibility in individuals.

[0288] Thus, based on the disclosure provided herein, those skilled in the art will understand that the dose and dosing regimen will be adjusted according to methods well known in the therapeutic field. That is, the maximum tolerated dose can be easily established, and the effective amount for providing a detectable therapeutic benefit to the patient can also be determined, as well as the time requirement for administering each agent to provide a detectable therapeutic benefit to the patient. Thus, although specific doses and dosing regimens are exemplified herein, these examples in no way limit the doses and dosing regimens that can be provided to a patient when implementing the present disclosure.

[0289] It should be noted that dosage values ​​may include single or multiple doses, which may vary with the type and severity of the condition to be alleviated. It should be further understood that for any particular subject, a particular dosage regimen should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the composition, and that the dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the embodied compositions. Furthermore, dosage regimens using the compositions of the present disclosure may be based on a variety of factors, including the type of disease, the patient's age, weight, sex, medical condition, the severity of the condition, the route of administration, and the particular antibody used. Thus, dosage regimens may vary widely, but can be routinely determined using standard methods. For example, doses may be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects, such as toxic effects and / or clinical laboratory values. Thus, the present disclosure encompasses intrapatient dose escalation as determined by the skilled artisan. Determining appropriate dosages and regimens is well known in the relevant art and will be understood to be encompassed by the skilled artisan once provided with the teachings disclosed herein.

[0290] The effective amount for tumor treatment can be measured by its ability to stabilize disease progression and / or ameliorate symptoms in patients, preferably by its ability to reverse disease progression, for example by reducing tumor size. The ability of the treatment or composition of the present disclosure to inhibit cancer can be evaluated, for example, by in vitro assays as described in the Examples, as well as in appropriate animal models that are predictive of efficacy in human tumors. The appropriate dosing regimen is selected to provide the optimal therapeutic response in each particular situation, for example, administered as a single bolus or continuous infusion, with possible adjustment of the dosage indicated by the exigencies of each case.

[0291] The treatments and compositions of the present disclosure may be administered by any method accepted in the art for administering peptides, proteins or antibodies, but are typically suitable for parenteral administration. As used herein, "parenteral administration" includes any route of administration characterized by physical disruption of the subject's tissue and administration through tissue disruption, thus generally resulting in direct administration into the bloodstream, muscle, or internal organs. Parenteral administration thus includes, but is not limited to, administration by injection, application through a surgical incision, application through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, subcutaneous, intraperitoneal, intramuscular, intrasternal, intracisternal, intravenous, intraarterial, intrathecal, intraurethral, ​​intracranial, intratumoral, and intrasynovial injection or infusion. Particular embodiments include intravenous and subcutaneous routes. In some embodiments, administration is by IV injection, e.g., IV infusion.

[0292] In some embodiments, the treatments and compositions of the disclosure may be administered according to the exemplary dosing regimens described in Example 10, for example, for parts 1a, 1b, 2a, and 2b of the described clinical trial.

[0293] For example, in certain embodiments, the anti-NKG2A antibody or antigen-binding portion may be administered at a dose of 8, 20, 100, 300, 750, or 1500 mg, or at a dose of 8-20, 20-100, 100-300, 300-750, or 750-1500 mg (e.g., as a monotherapy or as part of a combination therapy, as described herein). In certain embodiments, the anti-NKG2A antibody or antigen-binding portion is administered every 1, 2, 3, 4, 5, or 6 weeks. Further, the anti-NKG2A antibody or antigen-binding portion may be administered in a cycle of 7, 14, 28, 42, 56, 70, or 84 days.

[0294] In certain embodiments, the anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, may be administered (e.g., as part of a combination therapy described herein) at a dose of 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg, or at a dose of 50-100, 100-150, 150-200, 200-250, 250-300, 300-350, 350-400, 400-450, or 450-500 mg. In certain embodiments, the anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, may be administered every 1, 2, 3, 4, 5, or 6 weeks, and in certain embodiments, may be administered after 1, 2, 3, or 4 cycles of the anti-NKG2A antibody, or antigen-binding portion thereof.

[0295] In certain embodiments, the anti-EGFR antibody or antigen-binding portion thereof (e.g., a combination of two anti-EGFR antibodies or antigen-binding portions thereof) may be administered at a dose of 1, 3, 6, 9, 12, 15, or 18 mg / kg (e.g., as part of a combination therapy described herein). In some embodiments, the anti-EGFR or antigen-binding portion thereof may be administered at one of the doses, as a loading dose, at a different dose of the dose, as a maintenance dose, e.g., a loading dose of 9 mg / kg followed by a maintenance dose of 6 mg / kg. In certain embodiments, the anti-EGFR antibody or antigen-binding portion thereof is administered every 1, 2, 3, or 4 weeks.

[0296] In certain embodiments, the anti-HER2 antibody or antigen-binding portion thereof may be administered (e.g., as part of a combination therapy described herein) at a dose of 5, 10, 15, 20, 25, or 30 mg / kg. In certain embodiments, the anti-HER2 antibody or antigen-binding portion thereof is administered every 1, 2, 3, 4, 5, 6, 7, or 8 weeks.

[0297] Manufactured Products and Kits The present disclosure also provides articles of manufacture comprising an anti-NKG2A antibody, or antigen-binding portion thereof, described herein, and optionally an anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, described herein, and / or an anti-EGFR or anti-HER2 antibody, or antigen-binding portion thereof, described herein. For example, the articles of manufacture can include any of the therapeutic antibodies described herein, and can be for use in any of the methods of treatment described herein. Also provided are methods for making the articles.

[0298] The present disclosure also provides kits comprising an anti-NKG2A antibody, or antigen-binding portion thereof, described herein, and, optionally, an anti-PD-1 or anti-PD-L1 antibody, or antigen-binding portion thereof, described herein, and / or an anti-EGFR or anti-HER2 antibody, or antigen-binding portion thereof, described herein. For example, the kits can include any of the therapeutic antibodies described herein, and can be for use in any of the methods of treatment described herein.

[0299] The disclosure also provides articles of manufacture and kits that include one or more containers (e.g., single-use or multi-use containers) containing the treatment or composition described herein, optionally an additional biologically active molecule (e.g., another therapeutic agent), and instructions for use. The antibody or antigen-binding portion of the treatment or composition, and the optional additional biologically active molecule, can be packaged separately or together in any combination in suitable packaging, such as vials or ampoules made from non-reactive glass or plastic. In certain embodiments, the vials or ampoules hold concentrated stocks (e.g., 2x, 5x, 10x or more) of the antibody or antigen-binding portion and / or biologically active molecule. In certain embodiments, the articles of manufacture and kits include medical devices (e.g., syringes and needles) for administering the treatment or composition and / or the additional biologically active molecule; and / or suitable diluents (e.g., sterile water and saline).

[0300] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by those skilled in the art. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this disclosure. In the case of conflict, the present specification, including definitions, will control.

[0301] Generally, the nomenclatures used in connection with and techniques of cell and tissue culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, pharmaceutical and medicinal chemistry, and protein and nucleic acid chemistry and hybridization described herein are well known and commonly used in the art. Enzymatic reactions and purification techniques are performed according to manufacturer's specifications, as commonly accomplished in the art or as described herein.

[0302] Further, unless otherwise required by context, singular terms shall include the plural and plural terms shall include the singular. Throughout this specification and the embodiments, the terms "have" and "comprise", or variations such as "has", "having", "comprises", or "comprising", will be understood to imply the inclusion of a stated integer or group of integers, but not the exclusion of any other integer or group of integers.

[0303] All publications and other references mentioned herein are incorporated by reference in their entirety. Although a number of documents are cited herein, this citation does not constitute an admission that any of these documents form part of the common general knowledge in the art.

[0304] In order that this disclosure may be better understood, the following examples are set forth. These examples are for illustrative purposes only, and are not to be construed as limiting the scope of the disclosure in any way.

[0305] Abbreviation The abbreviations used herein are as follows: ADCC: Antibody-dependent cytotoxicity AE: Adverse event ALT: Alanine aminotransferase ANC: absolute neutrophil count AST: Aspartate aminotransferase AUC: Area under the plasma concentration-time curve BOIN: Bayesian Optimal Intervals CBR: Clinical profit rate Cmax: maximum plasma concentration CNS: central nervous system CPI: Checkpoint inhibitors CPS: Combined Positive Score CR: Completion response CRC: Colorectal cancer CTCAE: Common Terminology Criteria for Adverse Events DLT: Dose-limiting toxicity DOR: duration of response ECG: electrocardiogram ECOG: Eastern Cooperative Oncology Group EGFR: epidermal growth factor receptor HCC: hepatocellular carcinoma HER: Human epidermal growth factor receptor HLA: Human leukocyte antigen IHC: Immunohistochemistry IMP: Investigational medicinal product INR: International Normalized Ratio IRR: infusion-related reaction IS: Included Set ISH: in situ hybridization IV: Intravenous LVEF: left ventricular ejection fraction MAD: Maximum Administered Dose MSI: microsatellite instability MTD: maximum tolerated dose NCI: National Cancer Institute NE: Not assessable. NK: Natural killer (cells) NYHA: New York Heart Association ORR: Objective response rate OS: overall survival PBMC: Peripheral blood mononuclear cells PD: Progressive disease PFS: Progression-free survival PK: Pharmacokinetics PR: Partial response RECIST: Response Evaluation Criteria in Solid Tumors SD: stable disease SRC: Safety Review Committee ULN: upper limit of normal VPC: Visual predictive check WHO: World Health Organization WT: wild type EXAMPLES

[0306] Example 1: NK cell-mediated killing induced by mAb1 in selected cell lines expressing endogenous HLA-E This example describes the expression of endogenous HLA-E on the surface of tumor cell lines (HT-29, CCRF-CEM, A253, Detroit 562, CAL-120, FaDu) and the effect of mAb1 on NK cell-mediated killing of these tumor cell lines in vitro.

[0307] Materials and Methods Expression of endogenous HLA-E in six different cell lines (HT-29, CCRF-CEM, A253, Detroit 562, CAL-120, and FaDu) was examined by flow cytometry. Primary human NK cells isolated from healthy individuals were cloned for endogenous HLA-E (HLA-B * 0701 peptide) and treated with a single concentration of mAb1 or isotype control (IgG1 LALA). Calcein release was measured after 1.5 hours and % specific lysis was calculated.

[0308] result All six human tumor cell lines were shown to express endogenous HLA-E on their surface (Figure 1, Panel A). Treatment with mAb1 induced NK-mediated killing of these tumor cell lines compared to IgG1 LALA treatment (Figure 1, Panel B).

[0309] Example 2: Titration of mAb1 in comparison to BMS anti-NKG2A antibody analogs and monalizumab analogs This example describes the activity of mAb1 in comparison to both monalizumab analogs and BMS anti-NKG2A antibody analogs in a γδ T cell cytotoxicity assay. Primary, expanded human γδ T cells derived from healthy individuals were co-cultured with HLA-E expressing target cells (K562-HLA-E) and treated with the monalizumab analog mAb1, an analog of BMS anti-NKG2A.9 (IgG1.3f), BMS anti-NKG2A.11 (IgG1.3f), or an isotype control (IgG1-LALA).

[0310] Materials and Methods K562-HLA-E cells were incubated overnight with HLA-B * 0701 peptide. The next day, human primary γδ T cells derived from healthy individuals and expanded were isolated and incubated with two-fold titrations of the indicated antibodies starting at 50 μg / mL, followed by addition of calcein-loaded K562-HLA-E targets and incubation for 3 h. The killing capacity of γδ T cells was measured by calcein release into the supernatant. Specific lysis was calculated by subtracting spontaneous lysis (calcein-loaded 562-HLA-E cells only) and normalizing to maximum lysis (Triton X-100 lysis of calcein-loaded K562-HLAE cells).

[0311] result Direct comparison with either the monalizumab analogs (FIG. 2A) or the BMS anti-NKGA antibody analogs (NKG2A.9 and NKG2A.11) (FIG. 2B) demonstrated superior functional activity of mAb1.

[0312] Example 3: mAb1 enhances cetuximab-induced ADCC in FaDu cells This example describes the ability of mAb1 to enhance ADCC induced by EGFR-targeted antibodies in an NK cell cytotoxicity assay using target cells that endogenously express both HLA-E and EGFR. Titrations of cetuximab in combination with fixed concentrations of isotype control antibody, mAb1, or monalizumab analogs were first tested.

[0313] Materials and Methods FaDu cells were incubated overnight with HLA-B * 0701 peptide. The next day, human primary NKG2A+ γδ T cells derived from healthy individuals and expanded were co-cultured with FaDu cells (E:T ratio 1:10) and incubated with a 2-fold titration of cetuximab starting at 1 μg / mL in combination with the indicated antibodies at 25 ug / mL. After 90 minutes, the killing capacity of primary NK cells was measured by calcein release into the supernatant. Specific lysis was calculated by subtracting spontaneous lysis (calcein-loaded 562-HLA-E cells only) and normalizing to maximum lysis (Triton X-100 lysis of calcein-loaded K562-HLA-E cells).

[0314] result As shown in Figure 3, mAB1 substantially enhanced cetuximab-induced cytotoxicity. A more modest effect was observed when cetuximab was combined with the monalizumab analog. These data confirm that mAb1 can enhance ADCC-inducing antibody-mediated NK cell cytotoxicity.

[0315] Example 4: mAb1 enhances cetuximab- and futuximab / modotuximab-induced ADCC in A431 cells This example describes the effect of the combination of mAb1 and the ADCC-inducing monoclonal anti-EGFR antibodies treated with cetuximab or futuximab / modotuximab on the killing of tumor cell lines when co-cultured with primary NK cells.

[0316] Materials and Methods Human primary NKG2A+ NK cells from two healthy human individuals were cultured with calcein-labeled target cells (HLA-B * A431 HLA-E loaded with 0701 peptide + / EGFR + ) and treated with cetuximab or a combination of futuximab and modotuximab with or without mAb 1. Calcein release was measured after approximately 3 hours and % specific lysis was calculated.

[0317] result In combination with either cetuximab (Figure 4A) or the combination of futuximab and modotuximab (Figure 4B), mAb1 enhanced killing of the tumor cell line A431 over cetuximab or futuximab / modotuximab treatment alone or cetuximab + IgG1 LALA or futuximab / modotuximab + IgG1 LALA.

[0318] Example 5: mAb1 alone or in combination with cetuximab and futuximab / modotuximab induced NK cell activation (CD137 expression) This example describes the effect of mAb1, tested at one dose, alone or in combination with either cetuximab or futuximab / modotuximab, on inducing activation of primary NK cells in vitro. Expression of CD137 on NK cells, as a marker of their activation state, was examined by flow cytometry.

[0319] Materials and Methods A431 cells (HLA-E + / EGFR + ) overnight, *0701 peptide. The next day, NKG2A+ NK cells were isolated from fresh PBMCs from three healthy donors and co-cultured with A431 cells at a ratio of 10:1 in the presence of 10 ng / mL IL-2 and the antibodies or antibody combinations shown in Figure 5. After 48 hours of culture, cells were stained with Zombie Dye Live / Dead stain and anti-FcR antibodies, followed by surface staining with anti-CD3 APC-H7 (SK7, BD Biosciences), anti-CD56 BV650 (NCAM-16.2, BD Biosciences), anti-CD16 PE (B73.1, BD Biosciences), and anti-CD137 BV421 (B3, BD Biosciences) antibodies, and analyzed by flow cytometry using a FACScelesta.

[0320] result As shown in FIG. 5, mAb1 as well as cetuximab or futuximab / modotuximab induced NK cell activation alone, but CD137 expression was further induced in the combination of mAb1 with cetuximab or futuximab / modotuximab.

[0321] Example 6: mAb1 alone or in combination with futuximab / modotuximab induced secretion of IFNγ by NK cells This example describes the effect of mAb1, tested at one dose, alone or in combination with futuximab / modotuximab, on inducing IFNγ secretion by primary NK cells in vitro. Supernatants were harvested from treated co-cultures and analyzed by ELISA for IFNγ secretion.

[0322] Materials and Methods A431 cells (HLA-E + / EGFR + ) overnight, * The next day, the cells were pulsed with 0701 peptide. +NK cells were isolated from fresh PBMCs from three healthy donors and co-cultured with A431 cells at a ratio of 10:1 in the presence of 10 ng / mL IL-2 and anti-NKG2A antibody. After 48 h of culture, cell supernatants were harvested and IFNγ concentrations were quantified by ELISA (Invitrogen, 88-7316-88).

[0323] result mAb1 and futuximab / modotuximab treatment each stimulated the secretion of IFNγ alone, however, IFNγ secretion was further induced when the two were combined (FIG. 6).

[0324] Example 7: mAb1 enhances avelumab-induced ADCC in A431 and MDA-MB-231 tumor cell lines This example describes the effect of combining mAb1 with the ADCC-inducing monoclonal anti-PD-L1 antibody avelumab on killing of tumor cell lines when co-cultured with primary NK cells.

[0325] Materials and Methods Primary human NKG2A from two healthy individuals + NK cells were cultured using calcein-labeled A431 and MDA-MB-231 tumor cells (both HLA-B * The mice were co-cultured with 0701 peptide-loaded HLA-E+ / PD-L1+ mice. Calcein release was measured after approximately 3 hours, and the % specific lysis was calculated.

[0326] result In combination with avelumab, mAb1 enhanced killing of two tumor cell lines, A431 and MDA-MB-231, in both donors compared to avelumab treatment alone or velumab + IgG1 LALA.

[0327] Example 8: mAb1 in combination with avelumab induced NK cell activation (CD137) and IFNγ secretion This example describes the effect of a single dose of mAb1 alone or in combination with velumab on inducing CD137 expression and IFNγ secretion by primary NK cells in vitro.

[0328] Materials and Methods A431 cells (HLA-E + / EGFR + ) overnight, * The next day, the cells were pulsed with 0701 peptide. + NK cells were isolated from fresh PBMCs from healthy donors and co-cultured with A431 cells at a ratio of 10:1 in the presence of 10 ng / mL IL-2 and mAb1, avelumab, control antibody IgG1, a combination of mAb1 and avelumab, or a combination of velumab and control IgG1 LALA. After 48 hours of culture, cells were stained with Zombie Dye Live / Dead stain and anti-FcR antibodies, followed by surface staining with anti-CD3 APC-H7 (SK7, BD Biosciences), anti-CD56 BV650 (NCAM-16.2, BD Biosciences), anti-CD16 PE (B73.1, BD Biosciences), and anti-CD137 BV421 (B3, BD Biosciences) antibodies, and analyzed by flow cytometry using a FACScelesta. Supernatants from cell cultures were also harvested and the concentration of IFNγ was quantified by ELISA (Invitrogen, 88-7316-88).

[0329] result NK cell activation (as assessed by CD137 expression) was induced by mAb1 alone, but was further induced by the combination of mAb1 and avelumab (Figure 8A), and IFNγ secretion was induced by mAb1 alone, but was further induced by the combination of mAb1 and avelumab compared to avelumab treatment alone (Figure 8B).

[0330] Example 9: In vivo tumor growth inhibition by mAb1 in combination with avelumab This example demonstrates the in vivo efficacy of mAb1 in combination with avelumab against MDA-MB-231 tumor growth in CD34 humanized NOG mice.

[0331] Materials and Methods The human breast adenocarcinoma cell line, MDA-MB-231, was cloned into human CD34 + Tumors were implanted subcutaneously onto human IL15-boosted NOG mice humanized with umbilical cord blood stem cells. Tumors were measured in two dimensions with calipers three times a week, and tumor volumes (mm 3 ) was calculated according to the formula: (width)2 × length × 0.5. Treatments were randomly assigned to a tumor volume of 65 mm 3 Treatment was started at 0.5 mg / kg. Mice were treated by intraperitoneal injection of vehicle, mAb1 (10 mg / kg), avelumab (10 mg / kg), or a combination of mAb1 and avelumab (10 mg / kg per antibody), 3 times a week for a total of 9 treatments. A two-way ANOVA with Bonferroni's multiple comparison test was applied to compare tumor volumes at each time point between treatment groups. Statistical analysis was performed using GraphPad Prism version 9.1.0 (GraphPad Software, Inc.). At the end of the study, tumors were harvested and analyzed by flow cytometry. Cells were stained with anti-CD45-PE-Cy7, anti-CD3-FITC, anti-CD4-PE, and anti-CD8-APC-Cy7 antibodies, and Zombie Aqua was used for live / dead cell discrimination. Cells were analyzed using a FACSVerse flow cytometer and FacsDiva software. Data analysis was performed using GraphPad Prism version 9.1.0 (GraphPad Software, Inc.).

[0332] result As shown in Figure 9A, mAb1 in combination with avelumab demonstrated antitumor effects against MDA-MB-231 xenograft tumors implanted in CD34 humanized mice. Treatment induced a significant reduction in tumor growth (P<0.05 vs. vehicle control). Flow cytometric analysis of tumors demonstrated that CD4 + Compared with cells, CD8 + A higher percentage of cells are CD3 + An increase in cell invasion was evident (FIG. 9B).

[0333] Example 10: Phase 1a / 1b clinical protocol for mAb1 monotherapy and combination therapy This example describes a clinical investigational protocol for a Phase 1a / 1b, open-label, multicenter study investigating the safety, tolerability, and preliminary antitumor activity of mAb1 (anti-NKG2A) as monotherapy and in combination with mAb3 (anti-PD-1) in patients with advanced solid tumor malignancies. The study also includes an expansion portion with a triple combination of mAb1 and mAb3 and an anti-HER2 mAb or an anti-EGFR mAb (e.g., futuximab / modotuximab) in patients with metastatic gastric or colorectal cancer.

[0334] Maximum study duration for each participating patient: Each patient will participate in the study until confirmed disease progression, loss to follow-up, adverse events leading to discontinuation, significant non-compliance with the study protocol, withdrawal of consent, termination of the study, or death from any cause. For all patients, the maximum duration of treatment will not exceed 1 year for patients with a confirmed CR and 2 years for patients with a confirmed PR. Longer treatment durations may be permitted if, according to the investigator's judgment and after consultation with the sponsor, the patient's benefits outweigh the risks.

[0335] Study design Each of the four arms of the clinical trial is described below.

[0336] A. Part 1a - mAb1 Monotherapy The first primary objective is to evaluate the safety and tolerability of mAb1 as a single agent. The corresponding primary endpoints are: - incidence, severity, and relevance of AEs; - AEs leading to dose interruptions, delayed changes, and permanent treatment discontinuation; - Changes from baseline to the end of the study in safety laboratory values, vital signs, and ECG measurements.

[0337] The second primary objective was to determine the maximum tolerated dose (MTD) (or maximum administered dose [MAD]) of mAb1 as a single agent. The corresponding primary endpoints were: - The incidence of dose-limiting toxicities (DLTs) during Cycle 1, and - Incidence and severity of adverse events (AEs).

[0338] The first secondary objective is to evaluate the preliminary antitumor activity of mAb1 administered in cycle 1, followed by subsequent mAb3, according to investigator assessment using Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Corresponding secondary endpoints are: - ORR as assessed by investigator for antitumor activity (using RECIST v1.1); - Clinical benefit rate (CBR) (CR+PR+Stable disease (SD) ≥ 6 months); - Duration of response (DOR); - progression-free survival (PFS); and - Overall survival (OS).

[0339] The second secondary objective is to evaluate the immunogenicity of mAb1. The corresponding secondary endpoint is the formation of anti-mAb1 antibodies.

[0340] The third secondary objective is to characterize the pharmacokinetic (PK) profile of mAb1. Corresponding secondary endpoints include, but are not limited to, area under the plasma concentration-time curve (AUC), T max , maximum plasma concentration (C max ), and C troughmAb1 PK parameters including:

[0341] The first exploratory objective is to explore potential pharmacodynamic (PD) biomarkers of activity in tumor biopsies (pre- and post-treatment) and / or peripheral blood. Corresponding exploratory endpoints are: - NKG2A receptor occupancy by dose level in tumor biopsies and / or peripheral blood; - Alterations in gene expression profiles, immune cell subsets and activation status in tumor biopsies and / or peripheral blood; - changes in HLA-E and NKG2A expression in tumor biopsies and / or peripheral blood; and - Changes in plasma concentrations of soluble HLA-E and potentially other soluble factors.

[0342] The second exploratory objective is to explore potential PK / PD relationships through population modeling. The corresponding exploratory endpoint is the relationship between mAb1 PK and PD parameters and simulated outcomes in the PK / PD model.

[0343] The third exploratory objective is to evaluate potential predictive biomarkers of response to mAb1 from baseline tumor and / or peripheral blood samples. Corresponding exploratory endpoints are: - HLA-E expression and NKG2A positive immune infiltrate in tumor biopsies; - the plasma concentration of soluble HLA-E or potentially other soluble factors; and - Other potential predictive biomarkers in tumor biopsies associated with immunomodulatory agents or specific to the anti-NKG2A mechanism of action.

[0344] B. Part 1b - mAb1 + mAb3 combination therapy The first primary objective is to evaluate the safety and tolerability of mAb1 when administered in combination with mAb3. The corresponding primary endpoints are: - incidence, severity, and association of AEs; - Adverse events leading to dose interruptions, modifications, delays, and permanent discontinuation of treatment; and - Changes from baseline to the end of the study in safety laboratory values, vital signs, and ECG measurements.

[0345] The second primary objective is to determine the MTD or MAD and / or RP2D of mAb1 when administered in combination with mAb3. The corresponding primary endpoints are: - incidence of DLT during cycle 1; and - Incidence and severity of AEs.

[0346] The first secondary objective is to evaluate the preliminary antitumor activity of mAb1 in combination with mAb3 according to investigator assessment using RECIST v1.1. Corresponding secondary endpoints are: - ORR as assessed by investigator for antitumor activity (using RECIST v1.1); - Clinical profit rate (CBR) (CR+PR+SD≧6 months); - DOR; - PFS; and - OS.

[0347] The second secondary objective was to evaluate the immunogenicity of mAb1 alone or in combination with mAb3. The corresponding secondary objectives were: - formation of anti-mAb1 antibodies; and - Formation of anti-mAb3 antibodies.

[0348] The third secondary objective is to characterize the PK profile of mAb1 in combination with mAb3 and to explore potential PK interactions between mAb1 and mAb3. Corresponding secondary endpoints are: - Including but not limited to AUC, T max , C max , and C trough mAb1 PK parameters including; - External visual predictive checks (VPCs) performed on individual concentrations of mAb3 using an internal population PK model.

[0349] The first exploratory objective is to explore potential PD biomarkers of activity in combination with mAb3 in tumor biopsies (pre- and post-treatment) and peripheral blood. Corresponding exploratory endpoints are: - NKG2A receptor occupancy by dose level in tumor biopsies and / or peripheral blood; - Alterations in gene expression profiles, immune cell subsets and activation status in tumor biopsies and / or peripheral blood; - changes in HLA-E and NKG2A expression in tumor biopsies and / or peripheral blood; and - Changes in plasma concentrations of soluble HLA-E and potentially other soluble factors.

[0350] The second exploratory objective was to explore the relationship between PD-L1 tumor status (CPS) and response. The corresponding exploratory endpoints were: - PD-L1 expression in tumor biopsies; and - The relationship between PD-L1 expression in tumor biopsies and patient response.

[0351] The third exploratory objective is to explore any potential PK / PD relationships via population modeling that may support the selection of the RP2D. The corresponding exploratory endpoint is the relationship between mAb1 PK and PD parameters and simulated outcomes in the PK / PD model.

[0352] The four exploratory objectives are to evaluate potential predictive biomarkers of response to mAb1 in combination with mAb3 from baseline tumor and peripheral blood samples. The corresponding exploratory endpoints are: - HLA-E expression and NKG2A positive immune infiltrate in tumor biopsies; - plasma concentration of soluble HLA-E or other potential soluble factors; and - Other potential predictive biomarkers associated with immunomodulatory agents or specific to the anti-NKG2A mechanism of action in tumor biopsies.

[0353] C. Part 2a - mAb1 + mAb3 + margetuximab triple combination In this arm, margetuximab, an anti-HER2 monoclonal antibody, is added to the combination therapy. The primary objective is to evaluate the antitumor activity and efficacy of the triple combination (mAb1+mAb3+margetuximab) in HER2-positive patients with locally advanced unresectable or metastatic gastric cancer. The corresponding primary endpoint is ORR as assessed by investigator for antitumor activity using RECIST v1.1.

[0354] The first secondary objective is to evaluate the safety and tolerability profile of mAb1 in combination with mAb3 and margetuximab. Corresponding secondary endpoints are: - incidence and severity of AEs; - Adverse events leading to dose interruptions, modifications, delays, and permanent discontinuation of treatment; and - Changes from baseline to the end of the study in safety laboratory values, vital signs, and ECG measurements.

[0355] The second secondary objective is to confirm the RP2D of mAb1 in combination with mAb3 and margetuximab. Corresponding secondary endpoints are the overall safety profile, PK profile, and relationship between exposure and PD (i.e., safety, efficacy, and biomarkers).

[0356] The third secondary objective is to evaluate additional efficacy parameters to assess the antitumor activity of mAb1 in combination with mAb3 and margetuximab. Corresponding secondary endpoints are CBR, DOR, PFS, and OS.

[0357] The fourth secondary objective is to characterize the PK profile of mAb1 in combination with mAb3 and margetuximab and to investigate potential PK interactions between mAb1, mAb3 and margetuximab. Corresponding secondary endpoints are: - Including but not limited to AUC, T max , C max, and C trough mAb1 PK parameters including; - an external VPC performed for each concentration of mAb3 using an internal population PK model; and - External VPC performed for individual concentrations of margetuximab using literature population PK models.

[0358] The fifth secondary objective was to evaluate the immunogenicity of mAb1 in combination with mAb3 and margetuximab. The corresponding secondary endpoints were: - formation of anti-mAb1 antibodies; - formation of anti-mAb3 antibodies; and - Formation of anti-margetuximab antibodies.

[0359] The sixth secondary objective is to explore the relationship between PD-L1 (CPS) or HER2 tumor status and response. Corresponding secondary endpoints are: - PD-L1 and HER2 status in tumor biopsy; - The relationship between PD-L1 expression in tumor biopsies and patient response; and - Correlation between HER2 expression / amplification status in tumor biopsies and patient response.

[0360] The first exploratory objective is to further explore potential PD biomarkers of activity in combination with mAb3 and margetuximab and their relationship to PK and / or antitumor activity. Corresponding exploratory endpoints are: - NKG2A receptor occupancy in tumor biopsies and / or peripheral blood; - Alterations in gene expression profiles, immune cell subsets and activation status in tumor biopsies and / or peripheral blood; - Alterations in HLA-E and NKG2A expression in tumor biopsies and / or peripheral blood; - plasma concentrations of soluble HLA-E and potentially other soluble factors; and - Correlation between PD biomarkers and PK and / or patient response.

[0361] The second exploratory objective is to evaluate predictive biomarkers of response to mAb1 in combination with mAb3 and margetuximab from baseline tumor and peripheral blood samples and their relationship to antitumor activity. - HLA-E expression and NKG2A positive immune infiltrate in tumor biopsies; - plasma concentration of soluble HLA-E or potentially other soluble factors; - other potential predictive biomarkers associated with immunomodulatory agents or specific to the anti-NKG2A mechanism of action in tumor biopsies; and - Correlation between potential predictive biomarkers and patient response.

[0362] D. Part 2b - mAb1 + mAb3 + futuximab / modotuximab triple combination The primary objective is to evaluate the antitumor activity and efficacy of the triple combination (mAb1+mAb3+futuximab / modotuximab) in patients with metastatic colorectal cancer. The corresponding primary endpoint is ORR as assessed by investigator for antitumor activity using RECIST v1.1.

[0363] The first secondary objective is to evaluate the safety and tolerability profile of mAb1 in combination with mAb3 and futuximab / modotuximab. Corresponding secondary endpoints are: - incidence and severity of AEs; - Adverse events leading to dose interruptions, modifications, delays, and permanent discontinuation of treatment; and - Changes from baseline to the end of the study in safety laboratory values, vital signs, and ECG measurements.

[0364] The second secondary objective is to confirm the RP2D of mAb1 in combination with mAb3 and futuximab / modotuximab. Corresponding secondary endpoints are the overall safety profile, PK profile, and relationship between exposure and PD (i.e., safety, efficacy, and biomarkers).

[0365] The third secondary objective is to evaluate additional efficacy parameters to assess the antitumor activity of mAb1 in combination with mAb3 and futuximab / modotuximab. Corresponding secondary endpoints are CBR, DOR, PFS, and OS.

[0366] The fourth secondary objective is to characterize the PK profile of mAb1 in combination with mAb3 and futuximab / modotuximab and to investigate potential PK interactions between mAb1, mAb3 and futuximab / modotuximab. Corresponding secondary endpoints are: - Including but not limited to AUC, T max , C max , and C trough mAb1 PK parameters including; - An external VPC performed for each concentration of mAb3 using an internal population PK model; and - External VPC performed on individual concentrations of futuximab / modotuximab PK using an internal population PK model.

[0367] The fifth secondary objective was to evaluate the immunogenicity of mAb1 in combination with mAb3 and futuximab / modotuximab. Corresponding secondary endpoints were: - formation of anti-mAb1 antibodies; - formation of anti-mAb3 antibodies; and - Formation of anti-futuximab / modotuximab antibodies.

[0368] The sixth secondary objective was to explore the relationship between PD-L1 tumor status (CPS) and response. The corresponding secondary endpoints were: - PD-L1 status in tumor biopsy; and - Correlation between PD-L1 expression in tumor biopsies and patient response.

[0369] The first exploratory objective is to further explore potential PD biomarkers of activity in combination with mAb3 and futuximab / modotuximab and their relationship to PK and / or antitumor activity. Corresponding exploratory endpoints are: - NKG2A receptor occupancy in tumor biopsies and / or peripheral blood; - Alterations in gene expression profiles in tumor biopsies and / or peripheral blood; - Changes in immune cell subsets and activation status in tumor biopsies and peripheral blood; - Alterations in HLA-E and NKG2A expression in tumor biopsies and / or peripheral blood; - plasma concentrations of soluble HLA-E and potentially other soluble factors; and - Correlation between PD biomarkers and PK and / or patient response.

[0370] The second exploratory objective is to evaluate predictive biomarkers of response to mAb3 and mAb1 in combination with futuximab / modotuximab from baseline tumor and peripheral blood samples, and their relationship to antitumor activity. - HLA-E expression and NKG2A positive immune infiltrate in tumor biopsies; - plasma concentration of soluble HLA-E or potentially other soluble factors; - other potential predictive biomarkers associated with immunomodulatory agents or specific to the anti-NKG2A mechanism of action in tumor biopsies; and - Correlation between potential predictive biomarkers and patient response.

[0371] methodology This study will determine the safety and antitumor activity of mAb1 (anti-NKG2A) in combination with mAb3 (anti-PD1) (Part 1) and in triple combination with margetuximab or futuximab / modotuximab (Part 2) in two disease-specific cohorts in patients with advanced solid tumor malignancies.

[0372] To establish the MTD or MAD of mAb1 alone and in combination with mAb3, the safety of mAb1 monotherapy will be evaluated first in Part 1a and then in combination with mAb3 in Part 1b. The RP2D defined in Part 1b will be used in combination with margetuximab or futuximab / modotuximab in the dose expansion part (Part 2). The RP2D may be chosen at any time during Part 1b based on the overall safety profile, including PK / PD, whether or not the MTD has been characterized. The RP2D will not exceed the MTD (if characterized).

[0373] The trial will begin with a dose escalation of mAb1 monotherapy starting at a 20 mg dose (Part 1a). The DLT observation period will be 28 days (Cycle 1). Upon completion of the DLT evaluation period at the second dose level (100 mg), if deemed safe by the Safety Review Committee (SRC), the combination mAb1 (starting at 100 mg) + mAb3 (200 mg) dose escalation in Part 1b will begin. After this, escalation to the next mAb1 monotherapy dose level (300 mg) and beyond in Part 1a will be performed simultaneously with the combination dose escalation in Part 1b. There are five planned dose levels for the monotherapy dose escalation (20, 100, 300, 750, 1500 mg), plus an 8 mg dose de-escalation level, if needed. There are four planned dose levels for the combination dose escalation (100, 300, 750, 1500 mg), plus a 20 mg dose de-escalation level, if needed. mAb3 will be used as a fixed dose of 200 mg in the dose escalation of the Part 1b combination (see Table 2). [Table 2]

[0374] All patients in the mAb1 monotherapy dose escalation cohort in Part 1a will receive mAb3 (200 mg) monotherapy after completing one cycle of mAb1 and the DLT evaluation period. These patients will continue to receive mAb3 monotherapy until disease progression, unacceptable toxicity, or until 12 months after confirmed CR or 24 months after confirmed PR.

[0375] The triple combination addresses unmet medical needs (i.e., metastatic colorectal cancer and locally advanced unresectable or metastatic HER2 + The RP2D of the dual (mAb1+mAb3) in indications with monoclonal antibodies (gastric cancer) and potentially augmented by mAb1, such as monoclonal antibodies inducing ADCC (e.g., anti-HER2 margetuximab or other anti-HER2 mAbs and anti-EGFR mAbs futuximab / modotuximab), will be initiated as an expansion cohort (parts 2a and 2b). The RP2D of the mAb1+mAb3 combination will be combined with margetuximab (part 2a) or anti-EGFR (futuximab / modotuximab) (part 2b).

[0376] Part 1 (dose escalation) Part 1a will commence with a single-patient dose escalation (accelerated titration). The accelerated titration design will be switched to a Bayesian optimal interval (BOIN) design if a patient experiences a mAb1-related grade 2 AE or DLT. Up to three patients may be recruited as backfill patients to generate additional data to further define / characterize the RP2D and PK / PD relationships. The dose of mAb1 administered to backfill patients will be determined by the sponsor in agreement with the SRC and will not be higher than the MTD; these cohorts will follow the treatment schedule in Part 1a and will not be subject to a DLT window. Data from backfill patients will not contribute to the DLT analysis and will be considered in the overall safety analysis.

[0377] In Part 1b, 20 mg mAb1 or intermediate dose levels as well as tapers to higher dose levels may be explored. Patients at each mAb1 dose level will receive a mAb1 dose in combination with mAb3 (200 mg).

[0378] During Part 1, dosing will be staggered by at least 24 hours between all patients at each dose level. The first patient dosed at each dose level will be observed for safety for 24 hours; if no safety issues are noted, then a second patient will be enrolled at the same dose level and a third patient after another 24-hour safety observation. All patients enrolled in Parts 1a and 1b must have progressive or metastatic disease.

[0379] Other objectives of the study include preliminary evaluation of the antitumor activity of mAb1 in combination with mAb3, assessment of immunogenicity, characterization of the PK profile of mAb1 alone and in combination with mAb3, and evaluation of potential pharmacodynamic and other predictive biomarkers supporting mechanistic evidence in both blood and tumor tissue (tumor biopsies are optional in Part 1a and mandatory in Part 1b, with the exception of backfill patients).

[0380] Part 2 (dose expansion) Part 2 will be conducted in two expansion cohorts (Figure 10): (i) Part 2a in patients with HER2-positive metastatic gastric cancer with the addition of a third component anti-HER2 therapy (margetuximab); (ii) Part 2b in patients with metastatic colorectal cancer with the addition of a third component anti-EGFR therapy (futuximab / modotuximab).

[0381] Part 2a will be in patients with HER-2 positive gastric cancer who have failed first-line standard of care, including cytotoxic chemotherapy and trastuzumab and pembrolizumab, and are eligible to start second-line therapy. In patients with colorectal cancer (Part 2b), third and fourth lines of therapy will be allowed if the patient has previously received standard of care.

[0382] Part I Screening Criteria Patients are male or female patients aged ≥ 18 years. Medical and therapeutic criteria include: - Patients with histologically or cytologically confirmed unresectable locally advanced or metastatic solid tumor malignancies; - Patients with malignant lesions currently not amenable to surgical intervention, either due to medical contraindications or due to unresectable tumors; - Patients with measurable disease according to RECIST v1.1 and must have had radiological assessment of progression on previous therapy prior to study entry; - Patients who are resistant or intolerant to existing treatments known to provide clinical benefit or considered standard of care; - Estimated life expectancy ≥ 12 weeks. - ECOG performance status of 0 or 1; and - Documented radiological progression on previous line of treatment.

[0383] Inclusion criteria included: - Adequate hematological function based on the last assessment performed within 7 days prior to the first investigational medicinal product (IMP) dose, defined as: absolute neutrophil count (ANC) ≥ 1.5 x 10 9 / L; hemoglobin ≥ 8 g / dL (in the case of transfusion, hemoglobin assessment should be performed 2 weeks or more after transfusion); platelet count ≥ 75 × 10 9 / L; adequate coagulation function for all patients (adequate therapeutic INR levels should be confirmed for patients receiving anticoagulant therapy (excluding platelet antiaggregants)); - Adequate renal function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: creatinine clearance ≥ 40 mL / min, assessed using the Cockcroft & Gault formula; and - Adequate liver function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: total serum bilirubin ≤ 1.5 x upper limit of normal (ULN) (unless Gilbert's disease is confirmed), and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤ 3 x ULN (AST and ALT ≤ 5 x ULN, unless liver function abnormalities are due to underlying hepatic metastases).

[0384] Exclusion criteria included: - Patients with any other unresolved grade >1 toxicity associated with previous antineoplastic treatment, with the exception of persistent grade 2 alopecia or vitiligo, peripheral neuropathy and / or endocrine end-organ failure, adequately controlled with hormone replacement therapy; - Patients have had major surgery or have not recovered from side effects of surgery within 4 weeks prior to the first IMP dose. - Patients with any other serious / active / uncontrolled infection, infection requiring parenteral antibiotics, within 2 weeks prior to the first IMP dose; - Active hepatitis B virus infection determined as HBsAg positivity or active hepatitis C virus infection determined as detection of HCV RNA in serum or plasma by sensitive quantitative molecular methods; - HIV antibody carriers (patients with controlled HIV RNA are allowed if stable on medical therapy and undetectable by standard management); - Patients with a history of organ transplantation (e.g. stem cell transplant or solid organ transplant); - Patients with active thrombosis or a history of deep vein thrombosis or pulmonary embolism within 4 weeks prior to the first IMP dose who have not been adequately treated and are considered stable by the investigator; - Patients with active uncontrolled bleeding or known bleeding diathesis; - Patients with known clinically significant cardiovascular disease or conditions, including: need for antiarrhythmic drug therapy for ventricular arrhythmias or other uncontrolled arrhythmias (patients with uncontrolled atrial fibrillation (heart rate < 90) for > 30 days prior to enrollment); severe conduction disorders (e.g., third degree heart block); uncontrolled hypertension; congestive heart failure currently requiring treatment; class III or intravenous (IV) cardiovascular disease according to the New York Heart Association (NYHA) functional classification; history of acute coronary syndrome (including myocardial infarction and unstable angina), coronary angioplasty, stent placement, or bypass graft surgery within 6 months; - History of gastrointestinal perforation or intra-abdominal abscess within 28 days of enrollment; history of liver cirrhosis (defined as a known diagnosis of Child-Pugh B or C) or chronic liver disease; history of pulmonary fibrosis or related uncontrolled chronic pulmonary conditions; patients with non-healing wounds in any part of the body; - Patients who have previously received: small molecule inhibitors and / or other similar investigational agents: ≤ 2 weeks or 5 half-lives, whichever is shorter, chemotherapy, other monoclonal antibodies, antibody-drug conjugates, or other similar experimental treatments ≤ 3 weeks or 5 half-lives, whichever is shorter, or radioimmunoconjugates or other similar experimental treatments ≤ 6 weeks or 5 half-lives, whichever is shorter; - Patients must have resolved from an AE (from a previous anticancer drug treatment) to Common Terminology Criteria for Adverse Events (CTCAE) v5.0 grade 1 or less, with tolerable grade 2 neuropathy (patients receiving replacement hormone therapy due to a previous AE are not excluded from participation in the study if the associated AE resolved to grade 1 with the replacement therapy prior to the first dose on the study; - Patients who have previously received anti-NKG2A mAb; - Patients with known, untreated central nervous system (CNS) or leptomeningeal metastases, or spinal cord compression; patients with any of these not controlled by previous surgery or radiation therapy, or patients with symptoms suggestive of CNS involvement requiring treatment (only patients with clinically and radiologically stable CNS metastases for 4 weeks and with low corticosteroid treatment will be allowed to participate (≤10 mg / day prednisone or equivalent administered for 4 weeks will be allowed); - Other malignancies, including those that have been curatively treated and have been in remission for less than 5 years at screening. This exception to the minimum required remission period may apply for cervical intraepithelial neoplasia and basal cell skin cancer that are considered cured by sufficient treatment; - Treatment with systemic immunosuppressive therapy (except for steroids given in a prophylactic setting or in chronic low doses [≤10 mg prednisone equivalent / day]), inhaled, intranasal, intraocular, topical & intra-articular injections are permitted; - Prior radiation therapy if completed within 4 weeks prior to the first dose of IMP, unless given as a short course for symptomatic relief only; - Active autoimmune disease that may worsen if receiving immunostimulants (patients with type I diabetes, vitiligo, psoriasis, hypo- or hyperthyroidism not requiring immunosuppressive treatment are eligible); - Administration of a live vaccine within 28 days prior to enrollment; and - Known severe hypersensitivity (NCI CTCAE v. 5.0 grade ≥ 3) to the investigational products or any components in their formulations, including known severe hypersensitivity reactions to monoclonal antibodies.

[0385] Part 2a Screening Criteria Medical and therapeutic criteria include: - Histologically proven unresectable locally advanced or metastatic HER2 as determined by IHC as 3+ or 2+ by IHC and in situ hybridization (ISH-) amplification (≥ 2.0) in tumor biopsy collected at screening + Patients with cancer; - Patients must be eligible for second-line treatment and have been treated with first-line standard of care, including cytotoxic chemotherapy and trastuzumab and pembrolizumab; - Patients who received checkpoint inhibitor (CPI) treatment within the past 6 months must have documented confirmed radiological progression by iRECIST prior to study entry; - Patients with measurable disease according to RECIST v1.1 must have had radiological assessment of progression on prior therapy prior to study entry; - Estimated life expectancy ≥ 12 weeks. - ECOG performance status 0 or 1.

[0386] Inclusion criteria included: - Adequate hematologic function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: ANC ≥ 1.5 x 10 9 / L, hemoglobin ≥ 8g / dL, platelet count ≥ 75 × 10 9 / L, adequate coagulation function for all patients (adequate therapeutic levels of INR should be confirmed for patients receiving anticoagulant therapy (excluding platelet antiaggregants) that affect INR levels); - Adequate renal function based on the last assessment performed within 7 days prior to the first IMP dose, defined as creatinine clearance ≥ 30 mL / min, assessed using the Cockcroft & Gault formula; and - Adequate liver function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: total serum bilirubin < 1.5 x ULN (unless Gilbert's disease is confirmed), AST and alanine aminotransferase (ALT) ≤ 2.5 x ULN (unless liver function abnormalities are attributable to underlying liver metastases, AST and ALT ≤ 5 x ULN).

[0387] Exclusion criteria include those listed in Part 1 above, plus the following: - Left ventricular ejection fraction (LVEF) < 50% by echocardiogram or multi-gated acquisition scan; and - Any contraindications present in the margetuximab prescribing information.

[0388] Part 2b Screening Criteria Medical and therapeutic criteria include: - Patients must have histologically or cytologically confirmed adenocarcinoma of metastatic colorectal cancer (mCRC) (all other histological types are excluded), cannot undergo surgical intervention due to either medical contraindications or unresectability of the tumor, and have low microsatellite instability status according to institutional guidelines or guidelines from the College of American Pathologists; - Based on the ctDNA screening blood test analysis, patients should: (i) not have RAS (KRAS and NRAS) mutations in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4, and (ii) not have BRAF V600E mutations; - Patients who have received checkpoint inhibitor (CPI) treatment within the past 6 months must have documented confirmed radiological progression according to iRECIST prior to study entry; - Patients with measurable disease according to RECIST v1.1 must have had radiological evaluation of progression on prior therapy prior to study entry; - Life expectancy ≥ 12 weeks; and - ECOG performance status 0 or 1.

[0389] Inclusion criteria included: - Adequate hematologic function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: ANC ≥ 1.5 x 10 9 / L, hemoglobin ≥ 8 g / dL (in the case of transfusion, hemoglobin assessment should be performed 2 weeks or more after transfusion), platelet count ≥ 75 × 10 9 / L, and adequate coagulation function for all patients (adequate therapeutic levels of INR should be confirmed for patients receiving anticoagulant therapy (excluding platelet antiaggregating agents)); - Adequate renal function based on the last assessment performed within 7 days prior to the first IMP dose, defined as creatinine clearance ≥ 30 mL / min, assessed using the Cockcroft & Gault formula; - Adequate liver function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: total serum bilirubin < 1.5 x ULN (unless Gilbert's disease has been confirmed), and AST and alanine aminotransferase (ALT) ≤ 3.0 x ULN (AST and ALT ≤ 5 x ULN if liver function abnormalities are not due to underlying hepatic metastases); and - Serum potassium, serum phosphate, and serum magnesium within normal limits, with or without supplementation, based on the last assessment performed within 7 days prior to the first IMP dose.

[0390] Exclusion criteria included the following: - Patients with significant gastrointestinal abnormalities, including: grade >1 diarrhea at the time of enrollment and need for nutritional support; - Patients with grade >1 skin rash from prior anti-EGFR at the time of enrollment, or any other skin toxicity that would preclude participation in the study, as determined by the investigator; and - Known or suspected hypersensitivity to any of the excipients in formulated futuximab / modotuximab, cetuximab or panitumumab (grade 3 or 4 hypersensitivity reaction during previous treatment with either cetuximab or panitumumab).

[0391] Study drug The agents tested in this clinical trial are shown in Table 3 below: [Table 3]

[0392] Part 1 Drugs mAb1 is administered at the doses shown in Table 2 above, or adjusted if needed. Administration is via IV infusion every 2 weeks (Q2W) (± 1 day) on days 1 and D15 of each 28-day cycle. This 4-week (28-day) period constitutes one treatment cycle. mAb1 is infused over approximately 30 minutes. The duration of the infusion may be extended to 30 minutes or longer if indicated in the event of an infusion-related reaction (IRR). If a patient experiences an IRR during any cycle, the observation period should be extended to 2 hours during that cycle and all subsequent cycles for that patient.

[0393] mAb3 will be administered at a dose of 200 mg as an IV infusion every 2 weeks starting with C2D1 in part 1a or C1D1 in part 1b. If coadministered with mAb1 in part 1b, mAb1 will be administered first.

[0394] Part 2 Drugs Administration of mAb1 and mAb3 is as discussed above for part 1.

[0395] For Part 2a, margetuximab will be administered at 15 mg / kg every 3 weeks as a 120 minute (2 hour) IV infusion for the first dose, then over a minimum of 30 minutes for all subsequent doses. Margetuximab should not be administered as an IV push or bolus. Administration will be through an IV line containing a sterile, nonpyrogenic, low protein binding polyethersulfone (PES) 0.2 micron in-line or add-on filter. After the first 12 weeks, the investigator may decide to administer margetuximab on a Q4 weekly schedule.

[0396] For Part 2b, futuximab / modotuximab will be administered by IV infusion at a dose of 9 mg / kg on day 1 of cycle 1 (C1D1) (loading dose), then at a dose of 6 mg / kg weekly (± 2 days) starting on C1D8 for all subsequent administrations (maintenance dose). Dose adjustments should be made in the event of significant weight changes (± 10%) as measured at the beginning of the dosing cycle (CxD1). After the first 12 weeks, the investigator may decide to administer futuximab / modotuximab on a Q2W schedule.

[0397] Criteria for evaluation Standard response criteria will be applied for disease and response assessment (RECIST v1.1 and iRECIST v1). Assessments should be performed at intervals specified in the study schedule and at events of suspected progressive disease. The same disease assessment methods and techniques should be used as baseline throughout the study. Responses will be assessed by the investigator or qualified designee and described as CR, PR, SD, PD, or not evaluable (NE) at each endpoint. Disease assessments will be performed at baseline, at the end of cycle 2, and at the end of even-numbered cycles thereafter (approximately every 8 weeks unless a delay is required due to a delay in dosing), and at disease progression.

[0398] Per RECIST v1.1, responses should be confirmed by repeat imaging assessment within 4 weeks from the date the response was first documented. A scan to confirm the response may be performed as early as 4 weeks after the first documented response or at the next scheduled scan (8 week interval from the last scan), whichever is clinically indicated.

[0399] iRECIST will be used by investigators to assess tumor response and progression after first radiological progression according to RECIST v1.1; treatment decisions can be made accordingly.

[0400] Investigators should consider all lesions (target and non-target) when assessing tumor burden with repeat imaging before deciding whether to continue treatment. Patients will continue treatment (unless clinically unstable) until radiological confirmation of progression thereafter (at least 4 weeks after initial assessment of progression) as immune-related pseudoprogression may occur. Patients who are clinically unstable are not required to undergo repeat imaging for confirmation of progressive disease.

[0401] AEs will be graded using NCI CTCAE version 5.0. DLTs are defined as AEs that occur during the 28-day DLT observation period, are assessed as unrelated to disease progression, intercurrent illness, or concomitant medications or other etiology, and are considered to be related to IMP.

[0402] Pharmacokinetic measurements include: individual PK parameters, such as AUC or Cmax, are derived using a population PK modeling approach; and, an exploratory evaluation of the relationship between individual PK parameters and PD endpoints for efficacy or safety may be performed.

[0403] Pharmacodynamic assessments include: peripheral NKG2A receptor occupancy assessment and immunophenotyping, peripheral gene expression signature of target engagement, soluble HLA-E and potentially other soluble factors, and tumor biopsy for pharmacodynamic and predictive biomarker evaluation.

[0404] Example 11: In vivo efficacy of mAb1 in combination with anti-PD1 in a syngeneic MC38-HLAE tumor model in hNKG2A / hCD94 KI mice This example demonstrates the in vivo efficacy of mAb1 in combination with anti-PD1 mAb3 in hNKG2A / hCD94 KI mice (Biocytogen, China) engrafted with murine MC38-HLAE colon cancer cells.

[0405] Materials and Methods The mouse MC38-HLAE colon cancer cell line was subcutaneously implanted into a human NKG2A / CD94 double knock-in mouse model (Biocytogen, China). Treatment consisted of 100-mL IgG1-associated leukocytes with a tumor volume of 60 mm 3 Starting at 100 mg / kg, mice were treated three times a week by intraperitoneal injection of vehicle buffer, mAb1, anti-PD1 mAb3, or a combination of mAb1 and anti-PD1 mAb3 for a total of nine treatments (n=10 / group). All mAbs were dosed at 10 mg / kg. Tumors were measured in two dimensions by calipers three times a week, and tumor volumes (mm 3 ) was calculated according to the formula: (width)2 x length x 0.5.

[0406] Two-way analysis of variance with Bonferroni's multiple comparison test was applied to compare tumor volumes at each time point between treatment groups.

[0407] result As shown in FIG. 11, combination therapy with mAb1 and anti-PD1 mAb3 induced greater and earlier tumor eradication compared to single treatment with anti-PD1 mAb3.

[0408] Example 12: Titration of Trastuzumab with MIP-1b This example describes the effect of adding mAb1 to titrated trastuzumab on the expression of the pro-inflammatory cytokine MIP-1β in primary NK cells in vitro.

[0409] Materials and Materials WT SKOV3 cells were incubated overnight in a 30-well plate with HLA-B * The next day, these cells were pulsed with NKG2A peptide isolated from fresh PBMCs in the presence of 10 ng / ml IL-2 and anti-NKG2A antibodies. +The data show MIP-1β levels in cocultures with single donor NK cells. Similarly, HLA-E-transduced N87, BxPC3, SKOV3, A375, A549 and JIMT-1 cells for stable surface expression of HLA-E were incubated with 10 ng / ml IL-2 and anti-NKG2A and / or anti-HER2 antibodies or control antibodies in the presence of NKG2A cells isolated from fresh PBMCs. + NK cells were co-cultured at a ratio of 10:1. Data show MIP-1β levels in co-cultures with a single donor. After 48 hours of culture, supernatants were collected and MIP-1β concentrations were quantified by ELISA (Invitrogen, 88-7034-88).

[0410] result 25 μg / mL of mAb1 induced NK cell MIP-1β secretion when co-cultured with HLA-E+ cancer cells. Addition of titrated trastuzumab to IgG1 LALA (isotype control) antibody induced a dose-dependent induction of MIP-1β secretion, and this effect was increased when titrated trastuzumab was combined with mAb1 (Figure 12A). For all six cell lines transduced with HLA-E, there is a trastuzumab dose-dependent increase in MIP-1β when combining mAb1 with trastuzumab, even at low concentrations, compared to the control antibody IgG1 LALA (Figure 12B).

[0411] Example 13: Combined effects of dual PD-1 and NKG2A blockade on peripheral blood lymphocytes Materials and Methods Peripheral blood lymphocytes (PBLs) from three healthy donors were isolated by depleting CD14+ cells from peripheral blood mononuclear cells. 50,000 PBLs were seeded in round-bottom 96-well plates with 5,000 SKOV3 cells transduced with lentiviral vectors overexpressing HLA-E and PD-L1 and activated with trastuzumab 1 μg / mL or trastuzumab 1 μg / mL plus zoledronate 1 μM. PBLs were also treated with mAb1 (10 μg / mL), pembrolizumab (10 μg / mL), a combination of mAb1 (10 μg / mL) and pembrolizumab (10 μg / mL), or with the respective isotype controls IgG1 LALA (10 μg / mL), IgG4 (10 μg / mL), or a combination of both. Supernatants were collected after 3 days of incubation and concentrations of soluble mediators were measured by electrochemiluminescence using a multi-cytokine panel U-Plex kit (K151AEL-4, Mesoscale Discovery) on a Meso QuickPlex SQ120 plate reader (Mesoscale Discovery).

[0412] result PBLs were activated by trastuzumab alone, or by trastuzumab and phosphoantigens upregulated on the surface of SKOV3 cells by the addition of zoledronate, in the presence of HER2-positive SKOV3 cells engineered to overexpress HLA-E and PD-L1. The effect of PD-1 and NKG2A checkpoint inhibition was assessed individually or in combination in both activation contexts by measuring secretion of IFN-γ (a cytokine with multiple antitumor activities); granzyme B (mediates direct cytotoxicity); and MIP-1β / CCL4 (a proinflammatory chemokine). Trastuzumab activation, alone or in combination with mAb1 or pembrolizumab single agents, induced little or no secretion of soluble mediators. However, in the case of donor 1, the triple combination of trastuzumab, mAb1, and pembrolizumab resulted in a dramatic increase in the secretion of IFN-γ, granzyme B, and MIP-1β (Figure 13A). When PBLs were activated with both trastuzumab and zoledronic acid-induced phosphoantigen, single NKG2A blockade with mAb1 generally resulted in increased secretion of soluble mediators, whereas single PD-1 blockade had no or little effect compared to the corresponding single isotype controls. The combination of both mAb1 and pembrolizumab also resulted in an increase in IFN-γ, granzyme B, and MIP-1β secretion in all donors compared to mAb1 single treatment and compared to the combination of IgG1-LALA and IgG4 isotype controls. These results show the combined effect of NKG2A / PD-1 dual blockade in the context of trastuzumab stimulation.

[0413] Example 14: Phase 1b / 2 Clinical Protocol for mAb1 Combination Therapy This example describes an open-label, non-randomized, multi-arm, multi-center, Phase 1b / 2 study investigating the safety, tolerability, and anti-tumor activity of mAb1 in combination with pembrolizumab in MSI H / dMMR locally advanced unresectable or metastatic colorectal cancer (mCRC) and in the triple combination of mAb1, pembrolizumab, and trastuzumab in HER2-positive locally advanced unresectable or metastatic gastroesophageal junction (GEJ) and gastric adenocarcinoma (GA).

[0414] Maximum study duration for each participating patient: Each patient will participate in the study and have approximately 4 months of treatment plus 3 months of safety follow-up. The maximum duration of treatment for all patients will not exceed 2 years.

[0415] Study design Each of the two arms of the clinical trial, mAb1 + pembrolizumab (Figure 14) and mAb1 + pembrolizumab + trastuzumab (Figure 15), are described below.

[0416] The first primary objective is to evaluate the antitumor activity and efficacy of the combination of mAb1 and pembrolizumab in patients with MSI-H / dMMR (mismatch repair deficient) colorectal cancer (CRC) and the preliminary efficacy of the combination of mAb1, pembrolizumab, and trastuzumab in patients with HER2-positive gastroesophageal junction (GEJ) and gastric adenocarcinoma (GA) by assessing the overall response rate (ORR) by central review using Response Evaluation Criteria in Solid Tumors (RECIST) v1.1.

[0417] The second primary objective is to evaluate the safety and tolerability profile of the combination of mAb1 and pembrolizumab in patients with MSI-H / dMMR CRC and the safety and tolerability of the triple combination of S095029, pembrolizumab, and trastuzumab in HER2-positive gastroesophageal junction (GEJ) and gastric adenocarcinoma (GA).

[0418] The first secondary objective is to characterize the pharmacokinetic (PK) profile of mAb1 in combination with pembrolizumab in patients with CRC and in combination with pembrolizumab and trastuzumab in patients with HER2-positive disease.

[0419] The second secondary objective is to investigate potential PK / PD interactions between mAb1, pembrolizumab and trastuzumab in gastroesophageal and gastric cancer, and between mAb1 and pembrolizumab in colorectal cancer.

[0420] The third secondary objective is to continue evaluating the PK / PD profiles and further characterize the recommended Phase 2 doses (RP2D) of each combination.

[0421] A fourth secondary objective is to evaluate the immunogenicity of each antibody in the combination by assessing their potential for anti-drug antibody (ADA) formation.

[0422] A fifth secondary objective is to evaluate additional efficacy parameters and assess the antitumor activity of each combination.

[0423] methodology This study will determine the safety, tolerability, and antitumor activity of mAb1 in combination with pembrolizumab in MSI-H / dMMR locally advanced unresectable or mCRC, and the triple combination of mAb1, pembrolizumab, and trastuzumab in HER2-positive locally advanced unresectable gastroesophageal and gastric cancer.

[0424] mAb1 + pembrolizumab For the expansion Phase 2 cohort, patients will be treated with S095029 at RP2D via IV infusion on day 1 of each 21-day cycle every 3 weeks and pembrolizumab 200 mg via IV infusion on day 1 of each 21-day cycle every 3 weeks.

[0425] Screening Criteria Patients are male or female aged ≥ 18 years. Medical and therapeutic criteria include: - Patients must have histologically or cytologically confirmed locally advanced unresectable adenocarcinoma or mCRC; - Patients must have microsatellite instability-high status (MSI-H) or DNA mismatch repair deficient (dMMR) at the time of diagnosis according to institutional guidelines or according to the College of American Pathologists; - Patients must have progressed on at least two prior lines, including approved anti-PD(L)-1 agents such as pembrolizumab or nivolumab in combination with ipilimumab, and fluoropyrimidine-based standard of care chemotherapy regimens with oxaliplatin or irinotecan agents; - Patients with BRAF(V600E) mutations could have been included and treated with the BRAF inhibitor encorafenib in second-line; - Patients with measurable disease according to RECIST v1.1; - Patients must provide a suitable archival or freshly collected tumor biopsy; - Estimated life expectancy ≥ 12 weeks; - ECOG performance status 0 or 1.

[0426] Inclusion criteria included: - Adequate hematologic function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: ANC ≥ 1.5 x 10 9 / L, hemoglobin ≥ 8 g / dL. For transfusion, platelet count ≥ 75 × 10 9 / L, adequate coagulation function for all patients; - Adequate renal function based on last assessment, defined as creatinine clearance ≥ 30 mL / min; - Adequate liver function, defined as: total serum bilirubin <1.5 x ULN (<3 x ULN except in cases of confirmed Gilbert's disease), AST and ALT ≤3.0 x ULN; - Serum potassium, serum phosphate, and serum magnesium within normal limits, with or without supplementation, based on last evaluation.

[0427] Exclusion criteria included: - Major surgery within 4 weeks prior to first IMP dose; - Patients with other serious / active / uncontrolled infections; - Active hepatitis B virus infection; - HIV antibody carriers; - Patients with a history of organ transplantation; - Patients with active thrombosis or a history of deep vein thrombosis or pulmonary embolism; - Patients with active uncontrolled bleeding; - Patients with non-healing wounds on any part of the body; - Patients who have previously received anti-NKG2A mAb; - Patients with known, untreated central nervous system (CNS) disease; - treatment with systemic immunosuppressive therapy; - Prior radiation therapy if completed within 4 weeks prior to the first IMP dose; - Active autoimmune disease, which may worsen when receiving immune stimulants; - Administration of live vaccines within 28 days prior to inclusion; - Known severe hypersensitivity to the investigational products or any components in their formulations; - Any contraindications present in the pembrolizumab prescribing information.

[0428] mAb1 + pembrolizumab + trastuzumab Patients at the head of the cohort were treated with mAb1 at the recommended phase 2 dose (RP2D) via IV infusion on day 1 of each 21-day cycle every 3 weeks, trastuzumab at 6 mg / kg via IV infusion on day 1 of cycle 1, then every 3 weeks, and pembrolizumab 200 mg via IV infusion every 3 weeks. For the expansion phase 2 cohort, patients were treated with mAb1 at the RP2D via IV infusion on day 1 of each 21-day cycle every 3 weeks, trastuzumab at 8 mg / kg via IV infusion on day 1 of cycle 1, then every 3 weeks, and pembrolizumab 200 mg via IV infusion every 3 weeks.

[0429] Screening Criteria Patients are male or female aged ≥ 18 years. Medical and therapeutic criteria include: - Patients with locally advanced unresectable or metastatic gastroesophageal and gastric cancer; - Patients with histologically proven unresectable locally advanced or metastatic HER2+ cancer; - Patients who have progressed on two prior lines of treatment, including first-line and second-line standard of care regimens with a fluoropyrimidine and platinum-based combination, including trastuzumab with or without pembrolizumab; - Patients with measurable disease according to RECIST v1.1; - Estimated life expectancy ≥ 12 weeks; - ECOG performance status 0 or 1.

[0430] Inclusion criteria included: - Adequate hematological function based on the last assessment performed within 7 days prior to the first investigational medicinal product (IMP) dose, defined as: absolute neutrophil count (ANC) ≥ 1.5 x 10 9 / L, hemoglobin ≥ 8 g / dL, platelet count ≥ 75 × 10 9 / L; - Adequate coagulation function for all patients; - Adequate renal function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: creatinine clearance ≥ 30 mL / min; - Adequate liver function based on the last assessment performed within 7 days prior to the first IMP dose, defined as: serum total bilirubin ≤ 1.5 x upper limit of normal (ULN); aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤ 3 x ULN.

[0431] Exclusion criteria included: - Left ventricular ejection fraction (LVEF) <50% by echocardiogram; - Any existing contraindication to trastuzumab or pembrolizumab; - Major surgery within 4 weeks prior to the first IMP dose; - Patients with any other serious / active / uncontrolled infection; - Active hepatitis B virus infection; - HIV antibody carriers; - Patients with a history of organ transplantation; - Patients with active thrombosis or a history of deep vein thrombosis or pulmonary embolism; - Patients with active uncontrolled bleeding; - Patients with known clinically significant cardiovascular disease; - History of gastrointestinal perforation or intraperitoneal abscess; - History of cirrhosis or chronic liver disease; - History of pulmonary fibrosis or related uncontrolled chronic lung disease; - Patients with non-healing wounds on any part of the body; - Patients who have previously received anti-NKG2A mAb; - Patients with known untreated central nervous system (CNS) disease; - treatment with systemic immunosuppressive therapy; - Prior radiation therapy if completed within 4 weeks prior to the first IMP dose; - Active autoimmune disease.

[0432] Criteria for evaluation.

[0433] For both groups, tumor assessments will be performed every 9 weeks (+ / - 7 days) by medical imaging, and in case of response, confirmation will be required at least 4 weeks later. Patients will be followed up every 90 days (+ / - 7 days) for assessment of survival status after disease progression. Survival follow-up will last up to 2 years. [Table 4] TIFF2024537840000005.tif249169 TIFF2024537840000006.tif249169 TIFF2024537840000007.tif249169 TIFF2024537840000008.tif249169 TIFF2024537840000009.tif249169 TIFF2024537840000010.tif249169 TIFF2024537840000011.tif249169 TIFF2024537840000012.tif249169 TIFF2024537840000013.tif249169 TIFF2024537840000014.tif249169 TIFF2024537840000015.tif249169 TIFF2024537840000016.tif249169

Claims

1. a) an anti-NKG2A antibody or its antigen-binding portion, and an anti-PD-1 antibody or its antigen-binding portion; b) Anti-NKG2A antibody or its antigen-binding portion, and anti-PD-L1 antibody or its antigen-binding portion; c) Anti-NKG2A antibody or its antigen-binding portion, anti-PD-1 antibody or its antigen-binding portion, and anti-EGFR component; d) Anti-NKG2A antibody or its antigen-binding portion, anti-PD-1 antibody or its antigen-binding portion, and anti-HER2 antibody or its antigen-binding portion; e) Anti-NKG2A antibody or its antigen-binding portion, anti-PD-L1 antibody or its antigen-binding portion, and anti-EGFR component; or f) Anti-NKG2A antibody or its antigen-binding portion, anti-PD-L1 antibody or its antigen-binding portion, and anti-HER2 antibody or its antigen-binding portion A pharmaceutical composition for enhancing immunity in human patients in need thereof, including, Here, the anti-NKG2A antibody or its antigen-binding moiety competes or cross-competes with antibodies containing the heavy chain and light chain amino acid sequences of SEQ ID NOs. 9 and 10, respectively, or SEQ ID NOs. 19 and 20, respectively, for binding to human NKG2A, or binds to the same human NKG2A epitope as said antibody; and Herein, the anti-EGFR antibody component comprises one or two anti-EGFR antibodies or their antigen-binding moieties in the pharmaceutical composition.

2. The heavy chain complementarity determining regions (H-CDRs) 1-3 and light chain complementarity determining regions (L-CDRs) 1-3 of the anti-NKG2A antibody are a) Sequence IDs 1-6, respectively; or b) Sequence numbers 11-16 The pharmaceutical composition according to claim 1, comprising the amino acid sequence.

3. The heavy chain variable domain (VH) and light chain variable domain (VL) of the anti-NKG2A antibody are a) Sequence IDs 7 and 8, respectively; or b) Sequence IDs 17 and 18 The pharmaceutical composition according to claim 2, comprising the amino acid sequence.

4. The heavy chain (HC) and light chain (LC) of the anti-NKG2A antibody are a) Sequence IDs 9 and 10, respectively; or b) Sequence IDs 19 and 20, respectively The pharmaceutical composition according to claim 3, comprising the amino acid sequence.

5. The heavy chain complementarity determining regions (H-CDRs) 1-3 and light chain complementarity determining regions (L-CDRs) 1-3 of the anti-PD-1 antibody are a) Sequence numbers 21-26, respectively; b) Sequence IDs 31-36, respectively; c) Sequence IDs 41-46, respectively; d) Sequence numbers 51-56, respectively; e) Sequence numbers 61-66, respectively; or f) Sequence numbers 71-76 The pharmaceutical composition according to claim 1, comprising the amino acid sequence.

6. The heavy chain variable domain (VH) and light chain variable domain (VL) of the anti-PD-1 antibody are a) Sequence IDs 27 and 28, respectively; b) Sequence IDs 37 and 38, respectively; c) Sequence IDs 47 and 48, respectively; d) Sequence IDs 57 and 58, respectively; e) Sequence IDs 67 and 68, respectively; or f) Sequence IDs 77 and 78 The pharmaceutical composition according to claim 5, comprising the amino acid sequence.

7. The heavy chain (HC) and light chain (LC) of the anti-PD-1 antibody are a) Sequence IDs 29 and 30, respectively; b) Sequence IDs 39 and 40, respectively; c) Sequence IDs 49 and 50, respectively; d) Sequence IDs 59 and 60, respectively; e) Sequence IDs 69 and 70, respectively; or f) Sequence IDs 79 and 80, respectively The pharmaceutical composition according to claim 6, comprising the amino acid sequence.

8. The pharmaceutical composition according to claim 1, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, semiprimab, dostallimab, or retifanlimab.

9. The heavy chain complementarity determining regions (H-CDRs) 1-3 and light chain complementarity determining regions (L-CDRs) 1-3 of the anti-PD-L1 antibody are a) Sequence numbers 81-86, respectively; b) Sequence IDs 91-96, respectively; or c) Sequence numbers 101 to 106 The pharmaceutical composition according to claim 1, comprising the amino acid sequence.

10. The heavy chain variable domain (VH) and light chain variable domain (VL) of the anti-PD-L1 antibody are a) Sequence IDs 87 and 88, respectively; b) Sequence IDs 97 and 98, respectively; or c) Sequence IDs 107 and 108, respectively The pharmaceutical composition according to claim 9, comprising the amino acid sequence.

11. The heavy chain (HC) and light chain (LC) of the anti-PD-L1 antibody are a) Sequence IDs 89 and 90, respectively; b) Sequence IDs 99 and 100, respectively; or c) Sequence IDs 109 and 110, respectively The pharmaceutical composition according to claim 10, comprising the amino acid sequence.

12. The pharmaceutical composition according to claim 1, wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

13. The anti-EGFR component is a) Sequence numbers 111-116, respectively; b) Sequence numbers 121-126, respectively; c) Sequence IDs 131-136, respectively; or d) Sequence numbers 141-146 The pharmaceutical composition according to claim 1, comprising an anti-EGFR antibody or its antigen-binding moiety having heavy chain complementarity-determining regions (H-CDR) 1 to 3 and light chain complementarity-determining regions (L-CDR) 1 to 3 containing the amino acid sequence.

14. The anti-EGFR component is a) Sequence IDs 117 and 118, respectively; b) Sequence IDs 127 and 128, respectively; c) Sequence IDs 137 and 138, respectively; or d) Sequence IDs 147 and 148, respectively The pharmaceutical composition according to claim 13, comprising an anti-EGFR antibody or its antigen-binding moiety having a heavy chain variable domain (VH) and a light chain variable domain (VL) containing the amino acid sequence.

15. The anti-EGFR component is a) Sequence IDs 119 and 120, respectively; b) Sequence IDs 129 and 130, respectively; c) Sequence IDs 139 and 140, respectively; or d) Sequence IDs 149 and 150, respectively The pharmaceutical composition according to claim 14, comprising an anti-EGFR antibody having a heavy chain (HC) and a light chain (LC) containing the amino acid sequence.

16. The pharmaceutical composition according to claim 1, wherein the anti-EGFR component comprises an anti-EGFR antibody or its antigen-binding moiety having heavy chain complementarity-determining regions (H-CDRs) 1 to 3 and light chain complementarity-determining regions (L-CDRs) 1 to 3, each containing the amino acid sequences of SEQ ID NOs. 131 to 136, and an anti-EGFR antibody or its antigen-binding moiety having H-CDRs 1 to 3 and L-CDRs 1 to 3, each containing the amino acid sequences of SEQ ID NOs. 141 to 146.

17. The pharmaceutical composition according to claim 16, wherein the anti-EGFR component comprises an anti-EGFR antibody or its antigen-binding moiety having a heavy chain variable domain (VH) and a light chain variable domain (VL) containing the amino acid sequences of SEQ ID NOs. 137 and 138, respectively, and an anti-EGFR antibody or its antigen-binding moiety having VH and VL containing the amino acid sequences of SEQ ID NOs. 147 and 148, respectively.

18. The pharmaceutical composition according to claim 17, wherein the anti-EGFR component comprises an anti-EGFR antibody having a heavy chain (HC) and a light chain (LC) containing the amino acid sequences of SEQ ID NOs. 139 and 140, respectively, and an anti-EGFR antibody having an HC and LC containing the amino acid sequences of SEQ ID NOs. 149 and 150, respectively.

19. The pharmaceutical composition according to claim 1, wherein the anti-EGFR component is cetuximab, panitumumab, futuximab, modotuximab, or futuximab + modotuximab.

20. The heavy chain complementarity determining regions (H-CDRs) 1-3 and light chain complementarity determining regions (L-CDRs) 1-3 of the anti-HER2 antibody are a) Sequence numbers 151-156, respectively; or b) Sequence numbers 161-166 The pharmaceutical composition according to claim 1, comprising the amino acid sequence.

21. The heavy chain variable domain (VH) and light chain variable domain (VL) of the anti-HER2 antibody are a) Sequence IDs 157 and 158, respectively; or b) Sequence IDs 167 and 168, respectively The pharmaceutical composition according to claim 20, comprising the amino acid sequence.

22. The heavy chain (HC) and light chain (LC) of the anti-HER2 antibody are a) Sequence IDs 159 and 160, respectively; or b) Sequence IDs 169 and 170, respectively The pharmaceutical composition according to claim 21, comprising the amino acid sequence.

23. The pharmaceutical composition according to claim 20, wherein an anti-HER2 antibody or antigen-binding moiety is conjugated to DXd or DM1.

24. The pharmaceutical composition according to claim 1, wherein the anti-HER2 antibody is trastuzumab, margetuximab, trastuzumab dexrutecan, or trastuzumab emtansine.

25. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; and anti-PD-1 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 31-36, respectively; b) Anti-NKG2A antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; and anti-PD-1 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 37 and 38, respectively; or c) The pharmaceutical composition according to claim 1, comprising administering an anti-NKG2A antibody containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively, and an anti-PD-1 antibody containing the HC and LC amino acid sequences of SEQ ID NOs. 39 and 40, respectively.

26. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; Anti-PD-1 antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 61-66, respectively; and Anti-EGFR antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 131-136, respectively, and anti-EGFR components, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 141-146, respectively, and their antigen-binding moieties; b) Anti-NKG2A antibodies or their antigen-binding moieties, each containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; Anti-PD-1 antibodies or their antigen-binding moieties, each containing the VH and VL amino acid sequences of SEQ ID NOs. 67 and 68, respectively; and Anti-EGFR antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 137 and 138, respectively, and anti-EGFR antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 147 and 148, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; Anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 69 and 70, respectively; and Anti-EGFR components comprising anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 139 and 140, respectively, and anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 149 and 150, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

27. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 61-66, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 161-166, respectively; b) Anti-NKG2A antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 67 and 68, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 167 and 168, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 69 and 70, respectively; and anti-HER2 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 169 and 170, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

28. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 71-76, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 161-166, respectively; b) Anti-NKG2A antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 77 and 78, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 167 and 168, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 79 and 80, respectively; and anti-HER2 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 169 and 170, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

29. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 31-36, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 151-156, respectively; b) Anti-NKG2A antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 37 and 38, respectively; and anti-HER2 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 157 and 158, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 39 and 40, respectively; and anti-HER2 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 159 and 160, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

30. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; Anti-PD-1 antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 31-36, respectively; and Anti-EGFR antibodies or their antigen-binding moieties, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 131-136, respectively, and anti-EGFR components, each containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 141-146, respectively, and their antigen-binding moieties; b) Anti-NKG2A antibodies or their antigen-binding moieties, each containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; Anti-PD-1 antibodies or their antigen-binding moieties, each containing the VH and VL amino acid sequences of SEQ ID NOs. 37 and 38, respectively; and Anti-EGFR antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 137 and 138, respectively, and anti-EGFR antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 147 and 148, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; Anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 39 and 40, respectively; and Anti-EGFR components comprising anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 139 and 140, respectively, and anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 149 and 150, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

31. To the patient, a) Anti-NKG2A antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 1-6, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 31-36, respectively; and anti-EGFR antibodies or their antigen-binding moieties containing the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs. 111-116, respectively; b) Anti-NKG2A antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 7 and 8, respectively; anti-PD-1 antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 37 and 38, respectively; and anti-EGFR antibodies or their antigen-binding moieties containing the VH and VL amino acid sequences of SEQ ID NOs. 117 and 118, respectively; or c) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 39 and 40, respectively; and anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 119 and 120, respectively. A pharmaceutical composition according to claim 1, comprising administering the following:

32. The pharmaceutical composition according to claim 1, wherein the antibody or antigen-binding portion is administered to the patient simultaneously.

33. The pharmaceutical composition according to claim 1, wherein the antibody or antigen-binding portion is administered sequentially to the patient.

34. The pharmaceutical composition according to claim 1, wherein the patient has cancer.

35. The pharmaceutical composition according to claim 34, wherein the cancer is a hematological malignant lesion.

36. The pharmaceutical composition according to claim 34, wherein the cancer is a solid tumor.

37. The pharmaceutical composition according to claim 36, wherein the cancer is colorectal cancer, gastric cancer, or gastroesophageal cancer.

38. The pharmaceutical composition according to claim 1, wherein an anti-NKG2A antibody or its antigen-binding portion is administered in a dose of 8, 20, 100, 300, 750, or 1500 mg.

39. The pharmaceutical composition according to claim 38, wherein the antibody or portion is administered every two or three weeks.

40. The pharmaceutical composition according to claim 1, wherein an anti-PD-1 or anti-PD-L1 antibody or its antigen-binding portion is administered in a dose of 200 mg.

41. The pharmaceutical composition according to claim 40, wherein the antibody or portion is administered every two or three weeks.

42. The pharmaceutical composition according to claim 1, wherein an anti-NKG2A antibody or its antigen-binding portion is administered during a 21-day or 28-day cycle.

43. The pharmaceutical composition according to claim 1, wherein an anti-PD-1 or anti-PD-L1 antibody or its antigen-binding portion is administered simultaneously with or after one cycle of the anti-NKG2A antibody.

44. The pharmaceutical composition according to claim 1, wherein the anti-EGFR component is administered in a dose of 6 mg / kg, 9 mg / kg, or 6 mg / kg following a loading dose of 9 mg / kg.

45. The pharmaceutical composition according to claim 44, wherein the component is administered weekly or every two weeks.

46. The pharmaceutical composition according to claim 1, wherein an anti-HER2 antibody or its antigen-binding portion is administered at a dose of 15 mg / kg.

47. The pharmaceutical composition according to claim 46, wherein the antibody or portion is administered every three weeks or every four weeks.

48. The pharmaceutical composition according to claim 1, wherein the antibody or antigen-binding portion is formulated for intravenous injection.

49. A pharmaceutical composition for treating progressive solid tumor malignancies in a patient, comprising administering an anti-NKG2A antibody containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively, to the patient, wherein the antibody is administered by IV infusion every two weeks at doses of 8, 20, 100, 300, 750, or 1500 mg.

50. To the patient, a) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; and b) Anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs: 39 and 40, respectively. A pharmaceutical composition for treating progressive solid tumor malignant lesions in a patient, comprising administering the following: Anti-NKG2A antibody is administered by IV infusion at doses of 20, 100, 300, 750, or 1500 mg every three weeks, and A pharmaceutical composition comprising administering an anti-PD-1 antibody at a dose of 200 mg via IV infusion every three weeks, after completing a 21-day cycle of anti-NKG2A antibody administration.

51. To the patient, a) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; b) Anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 39 and 40, respectively; and c) Anti-HER2 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 159 and 160, respectively. This includes administering a drug to patients with metastatic HER2 ＋ A pharmaceutical composition for treating gastric cancer.

52. The pharmaceutical composition according to claim 51, wherein the cancer is locally advanced and / or unresectable.

53. The pharmaceutical composition according to claim 51, wherein the patient has failed first-line standard treatment.

54. Anti-NKG2A antibody is administered every two weeks in doses of 20, 100, 300, 750, or 1500 mg. After completing a 21-day cycle of anti-NKG2A antibody administration, anti-PD-1 antibody is administered at a dose of 200 mg every three weeks. Anti-HER2 antibodies are administered at a dose of 15 mg / kg every 3 or 4 weeks. The pharmaceutical composition according to claim 51, wherein the antibody is administered via IV infusion.

55. To the patient, a) Anti-NKG2A antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 9 and 10, respectively; b) Anti-PD-1 antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 69 and 70, respectively; and c) Anti-EGFR components comprising anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 139 and 140, respectively, and anti-EGFR antibodies containing the HC and LC amino acid sequences of SEQ ID NOs. 149 and 150, respectively. A pharmaceutical composition for treating metastatic colorectal cancer in a patient, comprising administering the following:

56. The pharmaceutical composition according to claim 55, wherein the patient has a low microsatellite instability state.

57. The pharmaceutical composition according to claim 55, wherein the patient is (i) free from RAS mutations in any of the following codons: codons 12 and 13 in exon 2, codons 59 and 61 in exon 3, and codons 117 and 146 in exon 4; and / or (ii) free from BRAF V600E mutations.

58. Anti-NKG2A antibody is administered every two weeks in doses of 20, 100, 300, 750, or 1500 mg. After completing a 28-day cycle of anti-NKG2A antibody administration, anti-PD-1 antibody is administered at a dose of 200 mg every two weeks. The anti-EGFR component is administered as a loading dose of 9 mg / kg, followed by doses of 6 mg / kg every one or two weeks. The pharmaceutical composition according to claim 55, wherein the antibody is administered by IV injection.

59. The pharmaceutical composition according to claim 1, further comprising administering to the patient radiotherapy, or at least one of a chemotherapeutic agent, an antitumor agent, and an anti-angiogenic agent.

60. The above procedure is as follows: a) Improved objective response rate; b) Improved clinical return; c) Duration of the improved response; d) Increased progression-free survival; and e) Increased overall survival A pharmaceutical composition according to any one of claims 34 to 59, which provides one or more of the following:

61. a) Human NKG2A and human PD-1; b) Human NKG2A and human PD-L1; c) Human NKG2A, human PD-1, and human EGFR; d) Human NKG2A, human PD-1, and human HER2; e) Human NKG2A, human PD-L1, and human EGFR; or f) Human NKG2A, human PD-L1, and human HER2 A multispecific antibody that specifically binds to a target.

62. a) an antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, and an antigen-binding domain of an anti-PD-1 antibody as defined in any one of claims 5 to 8; b) an antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, and an antigen-binding domain of an anti-PD-L1 antibody as defined in any one of claims 9 to 12; c) an antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, an antigen-binding domain of an anti-PD-1 antibody as defined in any one of claims 5 to 8, and an antigen-binding portion of one or two anti-EGFR antibodies as defined in any one of claims 13 to 16; d) an antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, an antigen-binding domain of an anti-PD-1 antibody as defined in any one of claims 5 to 8, and an antigen-binding moiety of an anti-HER2 antibody as defined in any one of claims 20 to 24; e) an antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, an antigen-binding domain of an anti-PD-L1 antibody as defined in any one of claims 9 to 12, and an antigen-binding moiety of one or two anti-EGFR antibodies as defined in any one of claims 13 to 16; or f) The antigen-binding domain of an anti-NKG2A antibody as defined in any one of claims 1 to 4, the antigen-binding domain of an anti-PD-L1 antibody as defined in any one of claims 9 to 12, and the antigen-binding portion of an anti-HER2 antibody as defined in any one of claims 20 to 24. A multispecific antibody according to claim 61, comprising:

63. The pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable excipient.

64. The pharmaceutical composition according to claim 63 for treating a human patient.

65. a) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8; b) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12; c) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-EGFR component as defined in any one of claims 13 to 19; d) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24; e) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-EGFR component as defined in any one of claims 13 to 19; or f) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24 A pharmaceutical composition for enhancing immunity in human patients requiring such enhancement, comprising an anti-NKG2A antibody as defined in any one of claims 1 to 4 or an antigen-binding portion thereof.

66. a) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8; b) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12; c) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-EGFR component as defined in any one of claims 13 to 19; d) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24; e) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-EGFR component as defined in any one of claims 13 to 19; or f) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24 A pharmaceutical composition for treating a human patient in combination with the above, comprising an anti-NKG2A antibody or its antigen-binding moiety as defined in any one of claims 1 to 4.

67. For the manufacture of pharmaceuticals to enhance immunity in human patients who require it, a) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8; b) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12; c) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-EGFR component as defined in any one of claims 13 to 19; d) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24; e) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-EGFR component as defined in any one of claims 13 to 19; or f) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24 The use of an anti-NKG2A antibody or its antigen-binding moiety as defined in any one of claims 1 to 4, in combination with the above.

68. For the manufacture of a pharmaceutical product for treating a human patient, a) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8; b) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12; c) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-EGFR component as defined in any one of claims 13 to 19; d) an anti-PD-1 antibody or its antigen-binding moiety as defined in any one of claims 5 to 8, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24; e) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-EGFR component as defined in any one of claims 13 to 19; or f) an anti-PD-L1 antibody or its antigen-binding moiety as defined in any one of claims 9 to 12, and an anti-HER2 antibody or its antigen-binding moiety as defined in any one of claims 20 to 24 The use of an anti-NKG2A antibody or its antigen-binding moiety as defined in any one of claims 1 to 4, in combination with the above.

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