Humanized anti-CLEC-1A antibody, its antigen-binding fragment, and its mimetic

Abstract

The present invention relates to the field of immunotherapy. The present invention provides novel specific humanized anti-CLEC-1A antibodies, their antigen-binding fragments and mimetics thereof, in particular antibodies. The compounds of the present invention can specifically bind to the CLEC-1A receptor and antagonize the binding of CLEC-1A to its endogenous ligand. The use of the compounds of the present invention can be useful in the treatment of deleterious conditions.

Description

[Technical Field]

[0001] The present invention relates to the field of immunotherapy. The present invention provides a novel, specific humanized anti-CLEC-1A antibody. The antibody of the present invention is capable of specifically binding to CLEC-1A and is an antagonist of human CLEC-1A, particularly antagonizing CLEC-1A binding to at least one of its ligands, particularly its endogenous ligand. The use of the compound of the present invention may be useful in the treatment of adverse conditions, including, but not limited to, cancer. [Background technology]

[0002] Immunotherapy treatments that harness a patient's immune system herald a new era of personalized medicine, offering hope for curative responses in patients with serious illnesses. Cellular immunity can eliminate or prevent diseases such as, but not limited to, cancer, autoimmune diseases, and allergic disorders. Recent advances in treatment, including cell engineering, disease targeting, and modulation of a patient's immune system, have led to more focused and effective responses to disease. Among these strategies, immunotherapy using immune checkpoint inhibitors or activators has become an essential weapon for the treatment of these diseases, particularly cancer. These molecules are frequently expressed by immune system cells, such as T cells or dendritic cells, but are also expressed by some cancer cells, enhancing immune responses against patients and maintaining or initiating immune cell responses against pathogenic cells. Immune checkpoints, which are inherently overly suppressive pathways in the immune system, are important for maintaining self-tolerance and minimizing collateral tissue damage.

[0003] C-type lectin receptors (CLRs) are a large family of transmembrane and soluble receptors that contain one or more carbohydrate recognition domains that can recognize a wide variety of glycans on pathogens or self-proteins. In these receptors, glycan recognition is mediated by Ca 2+ However, many related CLRs are dependent on Ca 2+These receptors are called C-type lectin-like receptors (CTLRs). These receptors are of particular interest due to their role in both innate and adaptive immunity. CTLRs are primarily expressed by myeloid lineage cells, such as monocytes, macrophages, dendritic cells (DCs), and neutrophils. CTLRs not only serve as antigen uptake receptors for internalization and presentation to T cells, but also trigger multiple signaling pathways, leading to NF-κB, type I interferon (IFN), and / or inflammasome activation. Due to their ability to present antigens and ensure a balance between cellular activation and inhibition, CTLRs have emerged as challenging pharmacological targets for treating a wide variety of diseases, including cancer, autoimmune diseases, and allergies. Although efforts to identify their endogenous ligands as well as to elucidate their role in immunity remain warranted, CTLR modulation appears to be a promising strategy for disease management.

[0004] Among these CTLRs, the CLEC1 receptor, CLEC1A receptor, and CLEC-1A receptor, also referred to by the acronyms CLEC1, CLEC1A, and CLEC-1A, are of particular interest. Although C-type lectin-like receptor-1 (CLEC-1) was identified several years ago, its downstream signaling and ligand remain uncharacterized. In humans and rodents, CLEC-1 is expressed by myeloid cells, such as monocytes, dendritic cells, and macrophages, but also by endothelial cells. CLEC-1 expression is downregulated by proinflammatory stimuli and upregulated by TGFβ. Interestingly, CLEC-1 has been found to be predominantly expressed intracellularly, particularly in human endothelial cells and neutrophils, suggesting that specific conditions are required for cell surface expression.

[0005] The present inventors have shown for the first time that CLEC-1A is expressed on the cell surface by conventional dendritic cells (cDCs) and by a small proportion of human blood monocytes and DCs, and is upregulated by the immunosuppressive cytokine TGFβ (see International Application Publication No. WO2018073440). The present inventors have shown that human CLEC-1A is expressed by M2-type tumor-promoting macrophages and by myeloid cells derived from pleural effusion melanoma and ovarian tumor ascites. In both humans and rodents, CLEC-1 has been demonstrated to act as an inhibitory receptor in myeloid cells, preventing IL12p40 expression and downstream Th1 and Th17 responses in vivo.

[0006] Furthermore, the use of anti-hCLEC-1A antibodies as CLEC-1A antagonists has been shown to increase human T cell proliferation and human IFN-gamma. Furthermore, mice lacking CLEC-1 have been demonstrated to be more resistant to tumor growth and exhibit increased survival rates in a mouse model of liver cancer. Therefore, CLEC-1A as a cell surface receptor may be a useful therapeutic tool for enhancing antitumor immunity in clinical settings. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2018073440 [Patent Document 2] U.S. Patent Publication No. 2007 / 0254295 [Non-patent literature]

[0008] [Non-Patent Document 1] Robles et al. (Blood advances, 2017) Summary of the Invention [Means for solving the problem]

[0009] In this regard, the inventors now provide for the first time humanized anti-CLEC-1A antibodies that recognize and specifically bind to the extracellular domain of human CLEC-1A and are antagonists of human CLEC-1A, in particular suitable for antagonizing the binding of CLEC-1A to at least one of its ligands, in particular endogenous ligands, and that, when used in vitro, correlate with a modulation, in particular an increase, of phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages. In a particular embodiment of the present invention, humanized anti-CLEC-1A antibodies are provided that recognize and specifically bind to the extracellular domain of human CLEC-1A and are antagonists of human CLEC-1A, in particular suitable for antagonizing the binding of CLEC-1A to at least one of its ligands, in particular endogenous ligands, and that, when used in vitro, correlate with a modulation, in particular an increase, of phagocytosis of tumor cells by macrophages.

[0010] As shown in the examples of the present invention, for the first time, humanized anti-CLEC-1A antibodies and antigen-binding fragments thereof and mimetics thereof, particularly anti-CLEC-1A antibodies, are provided that, when used in vivo and / or in vitro, correlate with modulation, particularly enhancement, of phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, particularly dendritic cells and / or macrophages. In contrast to the anti-CLEC-1A antibodies disclosed in the prior art (WO 2018 / 073440 and Robles et al., Blood Advances 2017) that bind to CLEC-1A and are used in some of the examples of the present invention and are antagonists of human CLEC-1A, it is demonstrated herein that humanized antibodies according to any of the embodiments of the present invention, when used in vitro, correlate with modulation, particularly enhancement, of phagocytosis of tumor cells by cells of the immune system. Tumor cells and / or secondary necrotic cells that interact with CLEC-1A escape phagocytosis by myeloid cells expressing CLEC-1A. The antibodies of the present invention interact with CLEC-1A in a manner that prevents functional interaction between CLEC-1A and tumor cells and / or secondary necrotic cells that normally interact with cells expressing CLEC-1A, and such interaction prevents tumor cell escape from phagocytosis. As exemplified herein, the antagonistic anti-CLEC-1A antibodies disclosed in the prior art (WO 2018 / 073440 and Robles et al. (Blood Advances 2017)) and used in some of the examples of the present invention do not correlate with modulation of tumor cell phagocytosis by myeloid cells, in particular by dendritic cells and / or macrophages. Modulation of tumor cell phagocytosis is merely illustrative (when antibodies according to the present invention are present in the examples). CLEC-1A-expressing myeloid cells, in particular CLEC-1A-expressing dendritic cells and / or macrophages, are not prevented from exerting their phagocytic ability, particularly by macrophages, of tumor cells and / or secondary necrotic cells, when an antibody, antigen-binding fragment thereof or mimetic thereof according to the invention is present. When the compounds of the invention are administered, several highly advantageous biological effects are achieved, in particular associated with the phagocytic ability of myeloid cells, including dendritic cells and / or macrophages.Antibodies of the present invention that are suitable antagonists of CLEC-1A correlate with modulation, particularly enhancement, of the phagocytic capacity of dendritic cells and / or macrophages, e.g., activated macrophages. Administration of anti-CLEC1A antibodies and antigen-binding fragments thereof, particularly anti-CLEC-1A antibodies, of the present invention correlates with enhanced phagocytosis of tumor cells and / or cancer cells and / or secondary necrotic cells by dendritic cells and / or macrophages by antagonizing the binding of CLEC-1A to its targets (at least one of its ligands) expressed by tumor cells and / or secondary necrotic cells. When macrophages or dendritic cells expressing CLEC-1A interact with cells expressing a CLEC-1A ligand, the phagocytic capacity of those macrophages or dendritic cells is inhibited or reduced. Tumor cells and secondary necrotic cells expressing a CLEC-1A ligand escape phagocytosis exerted by macrophages and dendritic cells. As shown in the examples of the present invention, when the anti-CLEC1A antibodies disclosed herein are administered, the inhibition of the phagocytic ability of macrophages and dendritic cells, particularly macrophages, is eliminated by antagonizing the interaction of CLEC-1A with tumor cells, thereby resulting in phagocytosis of tumor cells by macrophages and dendritic cells, particularly macrophages.

[0011] In addition to their effect on the phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, the humanized antibodies of the invention may further modulate, in particular improve or increase, T cell proliferation and / or T cell activation.

[0012] The antibodies described herein can be efficiently produced in recombinant production systems, making it possible to provide chimeric or (fully) humanized antibodies exhibiting the functional characteristics disclosed herein above in sufficient quantities for further development.

[0013] Furthermore, the humanized antibodies and antigen-binding fragments thereof and mimetics thereof, particularly antibodies, of the present invention have specific affinity for human CLEC-1A compared to their mouse orthologs and chimeric equivalents, because the antibodies of the present invention do not cross-react with mouse CLEC-1A protein in vitro. Furthermore, as shown in the examples of the present invention, the anti-CLEC-1A compounds of the present invention, particularly anti-CLEC-1A antibodies, specifically bind to the extracellular domain of CLEC-1A expressed on the plasma membrane of human cells in vitro.

[0014] In one embodiment of the present invention, the antibodies and antigen-binding fragments thereof and mimetics thereof of the invention disrupt the interaction of CLEC-1A, which is expressed by myeloid cells, in particular by dendritic cells and / or macrophages, with secondary necrotic cells and / or tumor cells, for example tumor cells present in a host having or expressing cancer, and / or with the intracellular contents of secondary necrotic cells and / or tumor cells. The inventors have found that ligands of CLEC-1A can be expressed or overexpressed by damaged or tumor cells (not necessarily on the membrane of those cells) and therefore can be involved in anti-tumor immunity and can ameliorate immune cell-induced tumor cell death.

[0015] Thus, the following: - specifically binds to human CLEC-1A, in particular CLEC-1A expressed on the plasma membrane of human cells; - an antagonist of human CLEC-1A, particularly suitable for antagonizing the binding of CLEC-1A to at least one of its ligands, in particular to one of its endogenous ligands, and having a superior antagonistic ability with respect to the binding of human CLEC-1A to one of its ligands than a chimeric antibody; - can be recovered in significant yields, allowing the provision of antibodies exhibiting the functional characteristics disclosed herein above in sufficient quantities for further development; and - when used in vivo and / or in vitro, correlates with a modulation, particularly an increase, of phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, particularly by dendritic cells and / or macrophages Humanized antibodies, antigen-binding fragments thereof and mimetics thereof for which we provide evidence are provided.

[0016] Such antibodies and their antigen-binding fragments and mimetics thereof are particularly suitable for their use in the prevention and / or treatment of several diseases or deleterious conditions in which the phagocytosis exerted by dendritic cells and / or macrophages needs to be improved, in particular for modulating the phagocytic activity of tumor cells and / or secondary necrotic cells, preferably by myeloid cells, in particular for improving the phagocytic capacity of dendritic cells and / or macrophages, and improving the outcome of the disease by increasing the phagocytosis of tumor cells by myeloid cells, in particular by dendritic cells and / or macrophages.

[0017] Such compounds may furthermore be particularly suitable for their use in the prevention and / or treatment of several diseases, in particular for modulating T cell responses, in particular by enhancing T cell activation and / or proliferation.

[0018] In one particular embodiment of the present invention, the humanized anti-CLEC-1A antibodies and their antigen-binding fragments and mimetics are suitable for reducing the total number of myeloid-derived suppressor cells, thereby resulting in a reduction of immunosuppressive cells, such as, but not limited to, immunosuppressive myeloid cells.

[0019] Thus, in a first aspect of the present invention, there is provided a method for treating a leukemia cell line comprising administering to a patient a therapeutically effective amount of ... an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 10, a VHCDR2 of SEQ ID NO: 11, and a VHCDR3 of SEQ ID NO: 12; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 13, a VLCDR2 of SEQ ID NO: 14, and a VLCDR3 of SEQ ID NO: 15; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 16, a VHCDR2 of SEQ ID NO: 17, and a VHCDR3 of SEQ ID NO: 18; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 19, a VLCDR2 of SEQ ID NO: 20, and a VLCDR3 of SEQ ID NO: 21; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 22, a VHCDR2 of SEQ ID NO: 23, and a VHCDR3 of SEQ ID NO: 24; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 25, a VLCDR2 of SEQ ID NO: 26, and a VLCDR3 of SEQ ID NO: 27, Antibodies or antigen-binding fragments thereof or mimetics thereof are disclosed.

[0020] The antibodies or antigen-binding fragments thereof or mimetics thereof according to this embodiment are suitable for antagonizing human CLEC-1A while exhibiting specific binding properties for its receptor. Furthermore, production in different cell lines, including but not limited to mammalian cell lines, is achieved in yields suitable for drug candidate development purposes. Furthermore, the antibodies or antigen-binding fragments thereof according to this embodiment may enhance phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, particularly dendritic cells and / or macrophages.

[0021] The present inventors have synthesized several anti-CLEC-1A humanized antibodies, each of which comprises a combination of heavy and light chain variable domains. Thus, in a second aspect of the present invention, the antibody heavy chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:3; and the antibody light chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:4, or the antibody heavy chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:5; and the antibody light chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:7, or the antibody heavy chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:6; and the antibody light chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO:7, or the antibody heavy chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO: 8; and the antibody light chain variable domain comprises or consists of the amino acid sequence set forth in SEQ ID NO: 9; Antibodies or antigen-binding fragments thereof are provided.

[0022] In another aspect, the present invention relates to a humanized antibody or an antigen-binding fragment thereof or a mimetic thereof that specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) and that, when used in vivo and / or in vitro, correlates with modulation of phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, particularly by dendritic cells and / or macrophages, particularly an increase of at least 10%, particularly at least 20% compared to a negative control.

[0023] In another aspect, the invention relates to a humanized anti-CLEC-1A antibody or an antigen-binding fragment thereof or a mimetic thereof as disclosed herein for use in the treatment of a disease or deleterious condition in which, inter alia, improved phagocytosis by dendritic cells and / or macrophages is required and / or improved phagocytic capacity of dendritic cells and / or macrophages is required to treat the disease or deleterious condition.

[0024] In another aspect, the present invention relates to the above-mentioned humanized anti-CLEC-1A antibodies and antigen-binding fragments and mimetics thereof for use in the prevention and / or treatment of diseases or disorders in which modulation of the phagocytic capacity of myeloid cells, in particular dendritic cells and / or macrophages, through modulation of phagocytosis of tumor cells and / or secondary necrotic cells, can improve the outcome of the disease or disorder, wherein the anti-CLEC-1A antibodies or antigen-binding fragments or mimetics thereof are antagonists of the interaction between human CLEC-1A and CLEC-1A ligand-expressing cells, in particular tumor or cancer cells and / or secondary necrotic cells expressing a CLEC-1A ligand. Such antibodies or antigen-binding fragments thereof can be identified using phagocytosis assays, including flow cytometry or microscopy, as described in the examples of the present invention. In a particular embodiment of the invention, the humanized antibody or its antigen-binding fragment or mimetic thereof may enhance phagocytosis of cancer cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular by at least 10%, in particular by at least 20% compared to a negative control. In a particular embodiment, phagocytosis may be assessed by the following experiment. Macrophages (MΦ) were generated from monocytes using M-CSF (100 ng / mL) for 5 days; then, the macrophages (MΦ) were preincubated with anti-CLEC1 compounds for 2 hours, and then non-Hodgkin's lymphoma (Raji; CD20+) and anti-CD20 mAb (rituximab) were added at 10 ng / mL each for 4 hours to provide an "Eat-me" signal. Phagocytosis analysis is performed by microscopy, and the percentage of phagocytosis is calculated based on the percentage of pHrodo (pHrodo-SE, Thermofisher) positive Raji cells among total macrophages.

[0025] In another aspect, the present invention relates to a humanized anti-CLEC-1A antibody, or an antigen-binding fragment thereof, or a mimetic thereof, as described above, for use in the prevention and / or treatment of a disease or disorder in which T cells have a deleterious effect, wherein the anti-CLEC-1A antibody, or an antigen-binding fragment thereof, or a mimetic thereof is an antagonist of the interaction between human CLEC-1A and secondary necrotic cells, and / or tumor cells, and / or tumor cells present in a host having or expressing cancer, and / or in the intracellular contents of permeabilized secondary necrotic cells and / or in the intracellular contents of permeabilized tumor cells.

[0026] In another aspect, the present invention relates to a method for increasing the phagocytic capacity of bone marrow cells, in particular dendritic cells and / or macrophages, comprising administering to a patient in need thereof an effective amount of a humanized anti-CLEC-1A antibody of the present invention or an antigen-binding fragment thereof or a mimetic thereof, in particular an anti-CLEC1A antibody, according to any embodiment disclosed herein; in particular, said anti-CLEC-1A antibody or an antigen-binding fragment thereof or a mimetic thereof is administered simultaneously, separately or sequentially with a conventional treatment or at least one second therapeutic agent, as defined herein.

[0027] In another aspect, the invention relates to a humanized anti-CLEC-1A antibody or an antigen-binding fragment thereof or a mimetic thereof as described above for use in the treatment of cancer, in particular for the treatment of liquid or solid cancer, and in particular for the treatment of lymphoma, colorectal cancer, mesothelioma or liver cancer.

[0028] Another aspect of the present invention relates to a combination of therapeutic compounds comprising a CLEC-1A antagonist antibody or its antigen-binding fragment or mimetic thereof, in particular an anti-CLEC-1A antibody, as a first therapeutic compound, and at least one second therapeutic compound selected from the group consisting of immunotherapeutic agents, in particular tumor-targeting antibodies, in particular anti-tumor-targeting antibodies suitable for activating and / or enhancing the phagocytic capacity of macrophages, in particular M1 macrophages, or chemotherapeutic agents, or radiotherapeutic agents. The inventors have shown that such combinations are particularly suitable for the treatment of cancer. As illustrated in the examples of the present invention, these combinations exert a synergistic effect in the treatment of cancer, resulting in a significant reduction in tumor growth, tumor volume, and / or improved survival rates.

[0029] Detailed Description of the Invention The phrase "secondary necrotic cells" or "cells undergoing secondary necrosis" therefore defines cells (including the cell lines disclosed herein) that have progressed to a stage of cellular change characterized by hypercondensed chromatin (pyknosis) and nuclear fragmentation (karyolysis), and possibly further features of cytoplasmic membrane rupture, release of activated caspase-3, and possibly cytoplasmic swelling and permeabilization of lysosomal membranes. Cells undergoing secondary necrosis are cells in which the apoptotic process has progressed to an autolytic necrotic event, i.e., the autolytic process of cell disintegration. The phrase "secondary necrotic cells" or "cells undergoing secondary necrosis" can also be appropriately defined in relation to markers of that specific stage of apoptotic cells, and markers that may further enable secondary necrotic cells to be distinguished from early apoptotic or primary necrotic cells are known and used. Such markers include annexin V and propidium iodide (PI) conjugated labels: early apoptotic cells are known to be annexin V positive and PI negative (annexin+ / PI-), while late apoptotic cells are known to be annexin V positive and PI positive, i.e., annexin / PI double positive (annexin+ / PI+). These markers are sometimes used in the art to indicate late apoptotic cells. As used herein, permeabilized cells are cells that allow access to intracellular or organelle antigens. Permeabilization allows entry of antibodies across the cell membrane, thereby allowing binding of the anti-CLEC1A antibodies, antigen-binding fragments, and mimetics thereof of the present invention to CLEC-1A, which is expressed in the intracellular compartment of the cell but not on the cell membrane, into the intracellular contents of those cells.

[0030] By "endogenous ligand" is understood a ligand that originates from the same species or in the same organism as the CLEC-1A receptor; for example, an endogenous human CLEC-1A ligand is the human ligand for the human CLEC-1A receptor; an endogenous mouse CLEC-1A ligand is the mouse ligand for the mouse CLEC-1A receptor.

[0031] The term "antibody" as used herein relates to a polyclonal antibody, a monoclonal antibody, or a recombinant antibody.

[0032] As used herein, "monoclonal antibody" is intended to refer to a preparation of antibody molecules, antibodies, that share a common heavy and light chain amino acid sequence, in contrast to "polyclonal" antibody preparations, which contain a mixture of antibodies of different amino acid sequences. Monoclonal antibodies can be produced by several known techniques, e.g., phage, bacterial, yeast, or ribosome display, as well as by classical methods exemplified by hybridoma-derived antibodies. Thus, the term "monoclonal" is used in reference to all antibodies derived from a single nucleic acid clone.

[0033] Antibodies of the present invention include recombinant antibodies. As used herein, the term "recombinant antibody" refers to antibodies produced, expressed, generated, or isolated by recombinant means, e.g., antibodies expressed using a recombinant expression vector transfected into a host cell; antibodies isolated from a recombinant combinatorial antibody library; antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes; or antibodies in which a particular immunoglobulin gene sequence (e.g., a human immunoglobulin gene sequence) has been assembled with other DNA sequences, produced, expressed, generated, or isolated in any other way. Recombinant antibodies include, for example, chimeric and humanized antibodies.

[0034] As used herein, "chimeric antibody" refers to an antibody in which variable domain sequences derived from the germline of a mammalian species, such as a mouse, have been grafted onto constant domain sequences derived from the germline of another mammalian species, such as a human.

[0035] As used herein, "humanized antibody" refers to antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0036] The antibody of the present invention is a humanized antibody. In one embodiment, the antibody of the present invention is a recombinant antibody. In one embodiment, the antibody of the present invention is a recombinant humanized antibody. The antibody of the present invention may be a deimmunized antibody. By "deimmunized", it is understood that the antibody shares a similar structure to the antibody of the present invention, but the structure of the antibody has been modified to remove known epitopes recognized by T cells in the antibody structure, thereby reducing the possibility of unwanted T cell responses.

[0037] As used herein, "antigen-binding fragment of an antibody" refers to a portion of an antibody, i.e., a molecule corresponding to a portion of the structure of an antibody of the present invention, which exhibits antigen-binding ability for CLEC-1A, possibly in its native form; such fragments in particular exhibit the same or substantially the same antigen-binding specificity for CLEC-1A compared to the antigen-binding specificity of the corresponding four-chain antibody. Advantageously, the antigen-binding fragment has a similar binding affinity to the corresponding four-chain antibody. However, antigen-binding fragments having reduced antigen-binding affinity relative to the corresponding four-chain antibody are also included within the scope of the present invention. Antigen-binding ability can be determined by measuring the affinity between the antibody and the target fragment. These antigen-binding fragments may also be referred to as "functional fragments" of antibodies.

[0038] As used herein, the term "mimetic" of an antibody refers to an antigen-binding antibody mimetic. Antigen-binding antibody mimetics are organic compounds that specifically bind to antigens but are not related to antibodies. They are typically artificial peptides or small proteins with a molar mass of approximately 3 to 20 kDa. Nucleic acids and small molecules are also sometimes considered antibody mimetics, but artificial antibodies, antibody fragments, and fusion proteins made from them are not. Common advantages over antibodies include better solubility, tissue penetration, heat and enzymatic stability, and relatively low production costs. Antibody mimetics are being developed as therapeutic and diagnostic agents. Antigen-binding antibody mimetics can also be selected from the group including affibodies, affilins, affimers, affitins, DARPins, and monobodies.

[0039] The term "CLEC-1" as used herein has its general meaning in the art and particularly relates to C-type lectin-like receptor-1 derived from mammalian species, particularly human CLEC-1. CLEC-1 belongs to the DECTIN-1 cluster of C-type lectin-like receptors (CTLRs), which also includes CLEC-2, DECTIN-1, CLEC-9A, MICL, MAH, and LOX-1.

[0040] The term "CLEC-1A" as used herein relates to CLEC-1A derived from a mammalian species, preferably human CLEC-1A. The reference sequence for human CLEC-1A corresponds to the sequence associated with accession number Q8NC01 Uniprot. Preferably, the term "human CLEC-1" or "human CLEC-1A" or "human CLEC-1 receptor" or "human CLEC-1A receptor" relates to the protein of amino acid sequence referenced by the Q8NC01 Uniprot accession number and encoded by the CLEC1A gene referenced by the NCBI accession number 51267. As used herein, the terms CLEC-1A, CLEC1A, CLEC1, CLEC-1, Clec1, Clec-1, Clec1A, and Clec-1A are used interchangeably and all refer to a mammalian CLEC1 receptor, its orthologous protein, or a homologous protein thereof, corresponding to the human CLEC-1A receptor corresponding to the sequence associated with accession number Q8NC01 Uniprot. In particular, CLEC-1A is a protein having the amino acid sequence of SEQ ID NO: 1. In particular, the extracellular domain of CLEC-1A is a protein having the amino acid sequence of SEQ ID NO: 2.

[0041] The term "CLEC-1 antagonist" as used herein has its general meaning in the art and relates to a compound, either natural or synthetic, that blocks, inhibits, or reduces the biological activity of CLEC-1. In particular, a CLEC-1 antagonist inhibits the interaction between CLEC-1 and at least one of its ligands. In particular, a CLEC-1 antagonist enhances T-cell responses, in particular increasing T-cell proliferation and / or cytokine synthesis such as IFN-gamma. Furthermore, the present invention relates to a compound, either natural or synthetic, that blocks, inhibits, or reduces the biological activity of CLEC-1. In particular, a CLEC-1 antagonist inhibits the interaction between the CLEC-1 receptor and at least one of its ligands, in particular all of its ligands. In particular, a CLEC-1 antagonist can bind to the CLEC-1 receptor or any one of its ligands.

[0042] As used herein, a "CLEC-1 antagonist" or "antagonist of CLEC-1" may correspond to a compound that binds to CLEC-1A and is selected from the group of antibodies, antigen-binding fragments of antibodies, antigen-binding mimetics of antibodies, antigen-binding fragments of antibodies or whole antibodies or mimetics.

[0043] The antagonistic potential of an antibody or its antigen-binding fragment or a mimetic thereof can be evaluated by suitable experiments as disclosed in the Examples of the present invention, in particular in Example 5. In particular, an antibody or its antigen-binding fragment or a mimetic thereof can be considered an antagonist of CLEC-1A, in particular of human CLEC-1A, if it (i) reduces the binding of the extracellular domain of CLEC-1A to secondary necrotic cells and / or tumor cells and / or to the intracellular contents of secondary necrotic cells, in particular to permeabilized RAJI cells and / or apoptotic PBMCs, compared to the same binding experiments in the absence of the antagonist antibody candidate, in particular if it reduces the binding of a fusion protein comprising the extracellular domain of the human CLEC-1A receptor fused to the Fc fragment of human immunoglobulin, in particular human IgG; and (ii) increases the phagocytosis of tumor cells by bone marrow cells compared to the same experiments in the absence of the antagonist compound. A decrease in binding is considered to be at least 1 log, particularly at least 2 log, particularly at least 3 log, compared to a negative experiment. An increase in phagocytosis of tumor cells is considered to be at least 10%, preferably at least 20%; most preferably at least 30%.

[0044] The antibodies and antigen-binding fragments of the present invention can be defined by structural characteristics: An antigen-binding fragment of an antibody is a fragment comprising its hypervariable domains, called CDRs (complementarity-determining regions), or a portion thereof comprising the recognition site for the antigen (i.e., the extracellular domain of CLEC-1A).

[0045] Each light and heavy chain variable domain (VL and VH, respectively) of a four-chain immunoglobulin has three CDRs, designated VLCDR1 (or LCDR1), VLCDR2 (or LCDR2), VLCDR3 (or LCDR3), and VHCDR1 (or HCDR1), VHCDR2 (or HCDR2), VHCDR3 (or HCDR3), respectively.

[0046] Those skilled in the art can determine the location of various regions / domains of an antibody relative to standard definitions in that regard, including reference numbering systems, as specified by reference to the KABAT numbering system, or by applying the IMGT "string of pearls" algorithm. In that regard, with regard to the definition of sequences in the present invention, it is specified that the boundaries of regions / domains may vary from one reference system to another. Thus, regions / domains defined in the present invention include sequences that exhibit approximately + / - 10% variation in length and position of the relevant sequence within the full-length sequence of an antibody variable domain.

[0047] In a specific embodiment of the present invention, the CDR domains of an antibody are designated by the Kabat nomenclature. In another specific embodiment of the present invention, the CDR domains of an antibody are designated by the IMGT nomenclature. In other words, some or all of the CDR domains of an antibody or antigen-binding fragment thereof of the present invention may be defined by the Kabat nomenclature; some or all of the CDR domains of an antibody or antigen-binding fragment thereof of the present invention may be defined by the IMGT nomenclature. In particular, all of the CDR domains of an antibody or antigen-binding fragment thereof of the present invention are defined by the Kabat nomenclature.

[0048] Therefore, based on the structure of the four-chain immunoglobulin, antigen-binding fragments can be defined by comparison with antibody sequences in available databases and prior art, in particular by comparison of the location of functional domains in those sequences, noting that the location of framework and constant domains is well defined for various classes of antibodies, in particular IgG, especially mammalian IgG, and such comparison also includes data on the three-dimensional structure of the antibody.

[0049] To illustrate specific embodiments of the present invention, antigen-binding fragments of antibodies containing variable domains comprising the antibody CDRs include Fv, dsFv, scFv, Fab, Fab', and F(ab')2. Fv fragments consist of the VL and VH domains of an antibody associated with each other by hydrophobic interactions; in dsFv fragments, the VH:VL heterodimer is stabilized by disulfide bonds; and in scFv fragments, the VL and VH domains are connected to each other via a flexible peptide linker, thereby forming a single-chain protein. Fab fragments are monomeric fragments obtained by papain digestion of antibodies and contain the entire L chain and the VH-CH1 fragment of the H chain, which are linked to each other by disulfide bonds. F(ab')2 fragments can be produced by pepsin digestion of antibodies below the hinge disulfide and contain two Fab' fragments and, additionally, a portion of the hinge region of an immunoglobulin molecule. Fab' fragments can be obtained from F(ab')2 by cleavage of the disulfide bond in the hinge region. F(ab')2 fragments are bivalent, i.e., contain two antigen-binding sites, like native immunoglobulin molecules; on the other hand, Fv (the VH:VL dimer comprising the variable region of Fab), dsFv, scFv, Fab, and Fab' fragments are monovalent, i.e., contain a single antigen-binding site. These basic antigen-binding fragments of the invention can be combined with each other to obtain multivalent antigen-binding fragments, such as diabodies, triabodies, or tetrabodies. Multivalent antigen-binding fragments are also part of the present invention.

[0050] As used herein, the term "bispecific" antibody refers to an antibody that recognizes two different antigens by having at least one region specific for a first antigen (e.g., derived from the variable region of a first antibody) and at least one second region specific for a second antigen (e.g., derived from the variable region of a second antibody). Bispecific antibodies are a type of multispecific antibody because they specifically bind to two target antigens. Multispecific antibodies that recognize two or more different antigens include, but are not limited to, antibodies that can be produced by recombinant DNA methods or produced chemically by any convenient method. Bispecific antibodies include any antibody, or conjugate of an antibody, or multimeric form of an antibody, that can recognize two different antigens. Bispecific antibodies include antibodies that have been reduced and modified to retain their bivalent character, and antibodies that have been chemically conjugated so that they may have several antigen recognition sites for each antigen, e.g., BiME (bispecific macrophage-enhancing antibody), BiTE (bispecific T-cell engager), DART (dual affinity retargeting); DNL (dock-and-lock).In particular, the bispecific antibodies according to the present invention can recognize and bind to CLEC-1A and comprise any combination of CDRs disclosed herein or any combination of heavy and light chain variable domains disclosed herein, and can exhibit the same functions and capabilities as the humanized anti-CLEC-1A antibodies of the present invention, and can also recognize and bind to CLEC-1A, including SIRP alpha, SIRP beta, SIRP gamma, CD47, CTLA-4, CD86 (B7.2), CD28, CD40, CD40L, ICOS, ICOS-L, OX40L, GITR, HVEM, BTLA, CD160, LIGHT, TNFRSF25, 2B4, CD48, Tim1, Tim3, Tim4, Gal9, LAG-3, CD40, CD40L, CD70, CD27, VISTA, B7H3, B7 and capable of recognizing and binding to at least one second compound selected from H4 (B7x), TIGIT, CD112, HHLA2 (B7-H7), TMIGD2 (CD28H), butyrophilin-like 2 (BTNL2), SIGLEC, AXL, B7.1, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, CD19, CD20, CD22, CD24, CD137 (4-1BB), CD137L (4-1BBL), CEA, CXCR3, CXCR4, EGFR, EGFRvIII, ELTD1, EMR1, EMR2, EMR3, EMR4P, ENG, EPCAM, EPHR, PD-L1, TLR1, TLR10, TLR2, TLR3, TLR4, VEGFR, VEGFR2, VIPR1, and VIPR2. A bispecific antibody may comprise two different paratopes, one recognizing human CLEC-1A corresponding to any combination of CDRs disclosed herein or any combination of heavy and light chain variable domains disclosed herein, and a second, different paratope recognizing another compound as listed herein above. Alternatively, the bispecific antibody may be a humanized anti-CLEC-1A antibody disclosed herein or an antigen-binding fragment thereof or a mimetic thereof linked to another compound selected from the lists herein above, or a fragment thereof, such as the extracellular domain of such a compound, if any.

[0051] All embodiments disclosed herein relating to antibodies are interchangeable mutatis mutandis to any compound according to the invention, in particular antigen-binding antibody fragments, antibody mimetics, in particular humanized recombinant antibodies.

[0052] In the following description of the invention, the term anti-CLEC-1A compound refers to either an antibody, or an antigen-binding fragment, or an antibody mimetic, whether recombinant or not, or a macromolecule comprising such an antibody or an antigen-binding fragment thereof. When the term anti-CLEC-1A antibody is used, the same compound is included by that term, except where specified in relation to a particular embodiment of the invention.

[0053] A "specific anti-CLEC-1A antibody or antigen-binding fragment thereof or mimetic thereof" is a compound that exhibits specific binding to CLEC-1A and not to other compounds, where binding is detectable in each case by methods known in the art, such as, but not limited to, Biacore analysis, Blitz analysis, ELISA assay, or Scatchard plot. Nevertheless, a specific "anti-CLEC-1A antibody or antigen-binding fragment thereof" may cross-react with compounds other than CLEC-1A, and the concept of specificity does not exclude that an antibody may cross-react with polypeptides other than CLEC-1A, but with lower affinity. Thus, a specific anti-CLEC-1A antibody or antigen-binding fragment thereof or mimetic thereof may also be defined as an antibody that exhibits high binding affinity to CLEC-1A but low binding affinity to other compounds.

[0054] Antibodies and antigen-binding fragments thereof In a first aspect, a compound that specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) is provided, which is: an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 10, a VHCDR2 of SEQ ID NO: 11, and a VHCDR3 of SEQ ID NO: 12; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 13, a VLCDR2 of SEQ ID NO: 14, and a VLCDR3 of SEQ ID NO: 15; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 16, a VHCDR2 of SEQ ID NO: 17, and a VHCDR3 of SEQ ID NO: 18; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 19, a VLCDR2 of SEQ ID NO: 20, and a VLCDR3 of SEQ ID NO: 21; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 22, a VHCDR2 of SEQ ID NO: 23, and a VHCDR3 of SEQ ID NO: 24; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 25, a VLCDR2 of SEQ ID NO: 26, and a VLCDR3 of SEQ ID NO: 27 Disclosed is a humanized antibody or antigen-binding fragment thereof or mimetic thereof, comprising: The CDRs located within the heavy and light chain variable domains of the exemplified antibodies are provided using Kabat numbering, and these combinations of CDR domains correspond to the CDR domains present on the heavy and light chains of each of the exemplified antibodies 11H11, 14H9, and 6C5, respectively.

[0055] In another aspect of the invention, there is provided an antibody that specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), comprising: an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 34, a VHCDR2 of SEQ ID NO: 35, and a VHCDR3 of SEQ ID NO: 36; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 37, a VLCDR2 of SEQ ID NO: 38, and a VLCDR3 of SEQ ID NO: 39; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 40, a VHCDR2 of SEQ ID NO: 41, and a VHCDR3 of SEQ ID NO: 42; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 43, a VLCDR2 of SEQ ID NO: 44, and a VLCDR3 of SEQ ID NO: 45; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 46, a VHCDR2 of SEQ ID NO: 47, and a VHCDR3 of SEQ ID NO: 48; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 49, a VLCDR2 of SEQ ID NO: 50, and a VLCDR3 of SEQ ID NO: 51 Disclosed is an antibody or antigen-binding fragment thereof or mimetic thereof comprising: The CDRs located within the heavy and light chain variable domains of the exemplified antibodies are provided using IMGT numbering, and these combinations of CDR domains correspond to the CDR domains present on the heavy and light chains of each of the exemplified antibodies 11H11, 14H9, and 6C5, respectively. In a specific embodiment of the present invention, a humanized antibody or antigen-binding fragment thereof or mimetic thereof that specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) comprises a combination of six CDR domains disclosed herein and shares at least 80%, particularly at least 85%, particularly at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of its framework regions with those of an antibody selected from 11H11, 14H9, and 6C5 and having the same six CDR domains. CDR domains may be defined by KABAT numbering or IMGT numbering. Framework regions correspond to amino acid residues located outside the CDR domains in the heavy and light chain variable domains.

[0056] Antibodies according to these embodiments (i.e., antibodies and antigen-binding fragments thereof having CDRs defined by Kabat numbering or IMGH numbering) are particularly suitable for enhancing phagocytosis of tumor cells by dendritic cells. Antibodies according to this definition have affinity for human CLEC-1A suitable for use in therapy, and at the same concentration, have a superior effect on the phagocytosis of tumor cells by dendritic cells compared to other anti-CLEC-1A antibodies, particularly compared to the control anti-CLEC-1A antibodies used in the examples of the present invention (see Figures 1 to 4). Furthermore, these antibodies and antigen-binding fragments elicit antagonistic potency against human CLEC-1A that is superior to that of their chimeric equivalents.

[0057] In one particular embodiment of the present invention, there is provided an antibody that specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), comprising: an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 10, a VHCDR2 of SEQ ID NO: 11, and a VHCDR3 of SEQ ID NO: 12; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 19, a VLCDR2 of SEQ ID NO: 20, and a VLCDR3 of SEQ ID NO: 21; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 10, a VHCDR2 of SEQ ID NO: 11, and a VHCDR3 of SEQ ID NO: 12; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 25, a VLCDR2 of SEQ ID NO: 26, and a VLCDR3 of SEQ ID NO: 27; an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 16, a VHCDR2 of SEQ ID NO: 17, and a VHCDR3 of SEQ ID NO: 18; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 25, a VLCDR2 of SEQ ID NO: 26, and a VLCDR3 of SEQ ID NO: 27; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 16, a VHCDR2 of SEQ ID NO: 17, and a VHCDR3 of SEQ ID NO: 18; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 13, a VLCDR2 of SEQ ID NO: 14, and a VLCDR3 of SEQ ID NO: 15; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 22, a VHCDR2 of SEQ ID NO: 23, and a VHCDR3 of SEQ ID NO: 24; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 19, a VLCDR2 of SEQ ID NO: 20, and a VLCDR3 of SEQ ID NO: 21; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 22, a VHCDR2 of SEQ ID NO: 23, and a VHCDR3 of SEQ ID NO: 24; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 13, a VLCDR2 of SEQ ID NO: 14, and a VLCDR3 of SEQ ID NO: 15 A humanized antibody or antigen-binding fragment thereof or mimetic thereof is provided, comprising:

[0058] In one particular embodiment of the present invention, there is provided an antibody that specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), comprising: an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 34, a VHCDR2 of SEQ ID NO: 35, and a VHCDR3 of SEQ ID NO: 36; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 43, a VLCDR2 of SEQ ID NO: 44, and a VLCDR3 of SEQ ID NO: 45; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 34, a VHCDR2 of SEQ ID NO: 35, and a VHCDR3 of SEQ ID NO: 36; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 49, a VLCDR2 of SEQ ID NO: 50, and a VLCDR3 of SEQ ID NO: 51; an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 40, a VHCDR2 of SEQ ID NO: 41, and a VHCDR3 of SEQ ID NO: 42; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 49, a VLCDR2 of SEQ ID NO: 50, and a VLCDR3 of SEQ ID NO: 51; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 40, a VHCDR2 of SEQ ID NO: 41, and a VHCDR3 of SEQ ID NO: 42; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 37, a VLCDR2 of SEQ ID NO: 38, and a VLCDR3 of SEQ ID NO: 39; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 46, a VHCDR2 of SEQ ID NO: 47, and a VHCDR3 of SEQ ID NO: 48; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 43, a VLCDR2 of SEQ ID NO: 44, and a VLCDR3 of SEQ ID NO: 45; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 46, a VHCDR2 of SEQ ID NO: 47, and a VHCDR3 of SEQ ID NO: 48; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 37, a VLCDR2 of SEQ ID NO: 38, and a VLCDR3 of SEQ ID NO: 39 A humanized antibody or antigen-binding fragment thereof or mimetic thereof is provided, comprising:

[0059] In one particular embodiment, the humanized antibody of the invention, or antigen-binding fragment thereof, or mimetic thereof, The amino acid sequence is - VHCDR1 of SEQ ID NO: 10; SEQ ID NO: 16, and SEQ ID NO: 22; - VHCDR2 of SEQ ID NO: 11; SEQ ID NO: 17, and SEQ ID NO: 23; and - VHCDR3 of SEQ ID NO: 12; SEQ ID NO: 18, and SEQ ID NO: 24 an antibody heavy chain variable domain comprising three VH CDRs selected from: The amino acid sequence is - VLCDR1 of SEQ ID NO: 13; SEQ ID NO: 19, and SEQ ID NO: 25; and - VLCDR2 of SEQ ID NO: 14; SEQ ID NO: 20, and SEQ ID NO: 26; and - VLCDR3 of SEQ ID NO: 15; SEQ ID NO: 21, and SEQ ID NO: 27 The antibody light chain variable domain comprises three VLCDRs selected from: Those CDRs located within the heavy and light chain variable domains of the exemplified antibodies are provided using Kabat numbering.

[0060] In one particular embodiment, the humanized antibody of the invention, or antigen-binding fragment thereof, or mimetic thereof, The amino acid sequence is - VHCDR1 of SEQ ID NO: 34; SEQ ID NO: 40, and SEQ ID NO: 46; - VHCDR2 of SEQ ID NO: 35; SEQ ID NO: 41, and SEQ ID NO: 47; and - SEQ ID NO: 36; SEQ ID NO: 42, and VHCDR3 of SEQ ID NO: 49 an antibody heavy chain variable domain comprising three VH CDRs selected from: The amino acid sequence is - VLCDR1 of SEQ ID NO: 37; SEQ ID NO: 43, and SEQ ID NO: 49; and - VLCDR2 of SEQ ID NO: 38; SEQ ID NO: 44, and SEQ ID NO: 50; and - VLCDR3 of SEQ ID NO: 39; SEQ ID NO: 45, and SEQ ID NO: 51 The antibody light chain variable domain comprises three VLCDRs selected from: Those CDRs located within the heavy and light chain variable domains of the exemplified antibodies are provided using Kabat numbering.

[0061] In one particular embodiment, the humanized antibody of the invention, or antigen-binding fragment thereof, or mimetic thereof, an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:3; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:4, or an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:5; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:7, or an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:6; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:7, or an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:8; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:9. Includes.

[0062] These combinations of heavy and light chain variable domains correspond, respectively, to the exemplary antibodies 11H11, the heavy and light chain variable domains of 14H9, a mutated version of the humanized heavy chain of 14H9 (where the mutations are located within the framework of the chain) and the light chain variable domain of 14H9, and 6C5. Antibodies according to this definition may be particularly suitable for modulating, in particular for improving, the phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells or macrophages, in particular in vitro and / or in vivo. Furthermore, antibodies according to this definition are particularly suitable, in particular compared to chimeric antibodies, for antagonizing the binding of human CLEC-1A to at least one of its ligands.

[0063] In one particular embodiment, the humanized antibody of the invention, or antigen-binding fragment thereof, or mimetic thereof, an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:3; SEQ ID NO:5; SEQ ID NO:6, and SEQ ID NO:8; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:4; SEQ ID NO:7, and SEQ ID NO:9 Includes.

[0064] The various antibody molecules and fragments may be derived from any of the commonly known immunoglobulin classes (isotypes), including, but not limited to, IgA, secretory IgA, IgE, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. In a specific embodiment of the invention, the variable region of the antibody may be associated with an antibody constant region, e.g., an IgG1, IgG2, IgG3, or IgG4 constant region. These constant regions may be further mutated or modified by methods known in the art to modify their binding ability to Fc receptors.

[0065] In one particular embodiment, the antibody or antigen-binding fragment or mimetic thereof according to the invention is a humanized monoclonal antibody, and the antibody light chain constant domain is derived from a human kappa light chain constant domain, in particular the light chain constant domain comprises or consists of the sequence of SEQ ID NO: 33.

[0066] In a specific embodiment, the antibody or antigen-binding fragment or mimetic thereof according to the invention is a humanized monoclonal antibody, and the antibody heavy chain constant domain is derived from a human IgG1, IgG2, IgG3, or IgG4 heavy chain constant domain, in particular the antibody heavy chain constant domain comprises or consists of the amino acid sequence of SEQ ID NO: 28 (human Fc IgG1), SEQ ID NO: 29 (human Fc IgG2), SEQ ID NO: 30 (human Fc IgG4), SEQ ID NO: 97 (Fc IgG1 N297A), SEQ ID NO: 100 (FcG1 LALA), and SEQ ID NO: 101 (Fc IgG1 LALAPG). Chimeric antibodies (e.g., those used as controls in the examples of the present invention) may comprise the heavy chain constant domain of SEQ ID NO: 31 (mouse FcG1) or SEQ ID NO: 32.

[0067] In other embodiments, the antibody or antigen-binding fragment or mimetic thereof binds to human CLEC-1A with an affinity of at least about 1x10-6M, 1x10-7M, 1x10-8M, 1x10-9M, 1x10-10M, 1x10-11M, 1x10-12M, or better, and / or binds to a target with an affinity that is at least 2-fold higher than its affinity for other compounds other than the human CLEC-1A receptor. In a particular embodiment, the antibody or antigen-binding fragment thereof binds to human CLEC-1A with an affinity constant (KD) of at least 1E-07M, particularly at least 1E-08M. In a particular embodiment, the antibody or antigen-binding fragment or mimetic thereof binds to human CLEC-1A with an affinity that is greater than 1-log, particularly greater than 2-log, and most preferably greater than 3-log, compared to the binding of a control anti-CLEC-1A antibody to CLEC-1a under the same binding conditions. The binding experiment can be carried out by any one of the binding experiments disclosed in the examples of the present invention.

[0068] In a particular embodiment, an anti-CLEC-1A antibody or its antigen-binding fragment or mimetic thereof according to the present invention is CLEC-1A-specific if the effective dose (ED50) of the compound to reach 50% of the maximum signal has an ED50 value for human CLEC-1A of less than 1500 ng / ml. ED50 can be determined by methods known in the art or by methods disclosed in the Examples of the present invention, for example, by cytometry as shown in Figure 11. In a particular embodiment, the binding between an anti-CLEC-1A antibody and human CLEC-1A, as defined herein above, can be considered specific if the effective dose (EC50) of the compound to reach 50% of the maximum signal in a binding assay is less than 1200 ng / ml, particularly less than 800 ng / ml, and in particular less than 400 ng / ml. Such ability can be assessed, for example, by methods exemplified in the Examples of the present invention.

[0069] In another particular embodiment, a specific anti-CLEC-1A antibody or antigen-binding fragment thereof or mimetic thereof according to the invention has an ED50 value (also referred to as EC50 value) for human CLEC-1A comprised between 1 ng / ml and 1000 ng / ml, particularly comprised between 5 ng / ml and 1500 ng / ml, particularly 800 ng / ml. EC50 can be determined by methods known in the art or by methods disclosed in the examples of the present invention, such as the method disclosed with respect to the data shown in Figure 11 and derived from Example 7.

[0070] The term "ED50" as used herein refers to the measurement of the effectiveness of a compound (e.g., an anti-CLEC-1A antibody or its antigen-binding fragment) in inducing 50% of a biological or biochemical function (e.g., a function or activity of CLEC-1A). For example, EC50 indicates how much of an anti-CLEC-1A antibody, its antigen-binding fragment, or a mimetic thereof is required to induce half of the activity of CLEC-1A. That is, EC50 is the 50% effective concentration (50% ED, or ED50) of an anti-CLEC-1A antibody, its antigen-binding fragment, or a mimetic thereof. ED50 is the concentration of a drug required for 50% efficacy in vitro. ED50 can be determined by techniques known in the art, for example, by constructing a dose-response curve to examine the effect of different concentrations of an anti-CLEC-1A compound on CLEC-1A binding to Fc-CLEC. Methods for this determination are disclosed, for example, in the Examples section of the present invention.

[0071] In the present invention, an anti-CLEC-1A antibody, or an antigen-binding fragment thereof, or a mimetic thereof can be considered to be an antagonist of CLEC-1A when it induces an increase in the KD value of an Fc-CLEC-1A protein for CLEC-1A by more than 1 log, preferably more than 2 log, more preferably more than 3 log, and most preferably more than 4 log in a binding competition assay in the presence of an antagonist antibody. This experiment can be performed by the Blitz method or ELISA under the experimental conditions exemplified in the examples of the present invention, for example.

[0072] Humanized antibodies, or antigen-binding fragments thereof, or mimetics thereof, can be obtained by substituting amino acid residues present in the constant regions of the variable chains (VH and / or VL) for human amino acid residues having the corresponding positions in human antibodies according to standard definitions and numbering, with the substitution level being 1% to 80%, more preferably 1% to 50%, even more preferably 1% to 20%, and especially 1% to 18% of the residues in the framework regions. The constant regions include, in particular, constant regions of framework regions (FR) defined in four-chain antibodies identified according to Kabat numbering or IMGT numbering, in particular according to Kabat numbering.

[0073] Anti-CLEC-1A antibodies can be humanized by known methods. For example, different combinations of CDRs disclosed herein can be grafted onto human heavy and / or light chain variable domains. Chimeric, humanized, and / or deimmunized antibodies of the present invention can belong to any immunoglobulin class, for example, unmodified antibodies. Preferably, they belong to the IgG subclass, for example, IgG1, IgG2, IgG3, or IgG4.

[0074] Methods for preparing recombinant antibodies (or antigen-binding fragments or mimetics thereof) by combining the variable regions of an antibody with a suitable linker or with the constant regions of other antibodies are well known in the art.

[0075] Furthermore, the present invention includes antibodies or antigen-binding fragments thereof or mimetics thereof, in particular humanized antibodies or antigen-binding fragments thereof, which compete with antibodies comprising the amino acid sequence of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6 or SEQ ID NO:8 as its variable heavy chain domain and the amino acid sequence of SEQ ID NO:4, SEQ ID NO:7 or SEQ ID NO:9 as its light chain variable domain, in particular the humanized antibodies 11H11, 14H9 or 6C5 which bind to the CLEC-1A receptor, as exemplified in the examples of the present invention, and which antagonize the binding of CLEC-1A to its target, and which enhance phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to negative controls, in particular compared to antibodies comprising the amino acid sequence of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6 or SEQ ID NO:8 as its variable heavy chain domain and the amino acid sequence of SEQ ID NO:4, SEQ ID NO:7 or SEQ ID NO:9 as its light chain variable domain.

[0076] In particular, the antibody of the present invention, or an antigen-binding fragment thereof, or a mimetic thereof, specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) and / or to an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 52, and further competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 3 and a light chain variable domain comprising or consisting of SEQ ID NO: 4, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 3 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33, and is an antagonist of human CLEC-1A, and is an antagonist of human CLEC-1A, in particular with an antibody comprising a heavy chain variable domain comprising or consisting of SEQ ID NO: 3 and a light chain variable domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 3 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 3 and a light chain domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33. It particularly antagonizes the binding of the extracellular domain of CLEC-1A, or of an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set out in SEQ ID NO: 52, to at least one of its ligands (particularly its target) which is particularly expressed by secondary necrotic cells and / or tumor cells, and particularly enhances the phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 3 and a light chain variable domain comprising or consisting of SEQ ID NO: 4, in particular compared to an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 3 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33.

[0077] In particular, the antibody of the present invention, or an antigen-binding fragment thereof, or a mimetic thereof, specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) and / or to an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 52, and further competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 5 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 5, and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, and is an antagonist of human CLEC-1A, and is an antagonist of human CLEC-1A, in particular with an antibody comprising a heavy chain variable domain comprising or consisting of SEQ ID NO: 5, and a light chain variable domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, in particular with an antibody comprising a heavy chain variable domain comprising or consisting of SEQ ID NO: 5, and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain variable domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33. It particularly antagonizes the binding of the extracellular domain of CLEC-1A, or of an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set out in SEQ ID NO: 52, to at least one of its ligands (particularly its target) expressed particularly by secondary necrotic cells and / or tumour cells, and particularly enhances phagocytosis of tumour cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 5 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular compared to an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 5, and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33.

[0078] In particular, the antibody of the present invention, or an antigen-binding fragment thereof, or a mimetic thereof, specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) and / or to an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 52, and further competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 6 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 6 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, and is an antagonist of human CLEC-1A, and is an antagonist of human CLEC-1A, in particular with an antibody comprising a heavy chain variable domain comprising or consisting of SEQ ID NO: 6 and a light chain variable domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, in particular with an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 6 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33. It particularly antagonizes the binding of the extracellular domain of CLEC-1A, or of an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set out in SEQ ID NO: 52, to at least one of its ligands (particularly its target) expressed particularly by secondary necrotic cells and / or tumor cells, and particularly enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 6 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular compared to an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 6 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33.

[0079] In particular, the antibody of the present invention, or an antigen-binding fragment thereof, or a mimetic thereof, specifically binds to the extracellular domain of human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor) and / or to an Fc-CLEC-1A protein comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 52, and further competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 8 and a light chain variable domain comprising or consisting of SEQ ID NO: 9, in particular an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 8 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101, and a light chain domain comprising or consisting of SEQ ID NO: 9 and SEQ ID NO: 33, and is an antagonist of human CLEC-1A, and is an antagonist of human CLEC-1A, in particular human CLEC-1A. and particularly enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 8 and a light chain variable domain comprising or consisting of SEQ ID NO: 9, in particular compared to an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 8 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 9 and SEQ ID NO: 33.

[0080] In a particular embodiment of the invention, the antibody or antigen-binding fragment or mimetic thereof may further enhance phagocytosis of cancer cells and / or secondary necrotic cells by myeloid cells, particularly dendritic cells and / or macrophages, compared to a negative control, particularly by at least 10%, particularly at least 20% compared to a negative control. In particular, the antibody or antigen-binding fragment or mimetic thereof, when used in vivo and / or in vitro, correlates with a modulation of phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, particularly dendritic cells and / or macrophages, particularly an increase compared to a negative control, particularly an increase in phagocytosis of tumor cells and / or secondary necrotic cells compared to a negative control, particularly by at least 10%, particularly at least 20%.

[0081] Cross-competing antibodies (or compounds) and antibodies (or compounds) recognizing the CLEC-1A receptor can be identified using routine techniques, e.g., immunoassays, e.g., by demonstrating the ability of one antibody to block the binding of another antibody to a target antigen, e.g., by competitive binding assays. Competitive binding can be determined using assays such as those described in the Examples of the present invention. In particular, competitive binding can be determined using the method illustrated in Example 7, in which the interaction and competition of antibodies for His-CLEC1 is tested by ELISA. Cross-competition exists when the anti-CLEC-1A compound being tested reduces the binding of the other antibody by at least 50%, at least 60%, particularly at least 70%, and particularly at least 80% (or vice versa) compared to a positive control lacking one of the antibodies (or compounds).

[0082] The present invention further relates to genetic constructs encoding at least a portion of the specific anti-CLEC-1A antibodies and antigen-binding fragments thereof and mimetics thereof described herein.

[0083] To that end, the present invention further relates to a nucleic acid molecule or a combination of nucleic acid molecules encoding an antibody or antigen-binding fragment thereof or a mimetic thereof according to any one of the definitions disclosed herein, in other words, the nucleic acid molecule encodes at least six CDR domains of an antibody or antigen-binding fragment thereof or a mimetic thereof.

[0084] The present invention may further relate to a combination of a first nucleic acid molecule encoding at least one variable heavy chain domain of an antibody and a second nucleic acid molecule encoding at least one variable light chain domain of an antibody, wherein the combination of the first and second nucleic acid molecules encodes at least six CDR domains of an antibody or antigen-binding fragment thereof or mimetic thereof according to any embodiment disclosed herein.

[0085] In one particular embodiment of the invention, a nucleic acid molecule or combination of nucleic acid molecules encoding an antibody or antigen-binding fragment thereof or a mimetic thereof according to any one of the definitions disclosed herein, comprising: (a) the nucleic acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 68, or SEQ ID NO: 69 encoding CDR1 of the light chain variable domain; and / or (b) the nucleic acid sequence of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 70, or SEQ ID NO: 71 encoding CDR2 of the light chain variable domain; and / or (c) the nucleic acid sequence of SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 72, or SEQ ID NO: 73 encoding the CDR3 of the light chain variable domain; and / or (d) the nucleic acid sequence of SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 89, or SEQ ID NO: 90 encoding CDR1 of the heavy chain variable domain; and / or (e) the nucleic acid sequence of SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 91, or SEQ ID NO: 92 encoding CDR2 of the heavy chain variable domain; and / or (f) the nucleic acid sequence of SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 93, or SEQ ID NO: 94 encoding the CDR3 of the heavy chain variable domain There is provided a nucleic acid molecule or combination of nucleic acid molecules comprising or consisting of:

[0086] In one particular embodiment of the invention, a nucleic acid molecule or combination of nucleic acid molecules encoding an antibody or antigen-binding fragment thereof or a mimetic thereof according to any one of the definitions disclosed herein, comprising: a) the nucleic acid sequence of SEQ ID NO: 53, SEQ ID NO: 60, or SEQ ID NO: 67 encoding the variable domain of the light chain; and / or b) the nucleic acid sequence of SEQ ID NO: 74, SEQ ID NO: 81, SEQ ID NO: 88, or SEQ ID NO: 95 encoding the variable domain of the heavy chain There is provided a nucleic acid molecule or combination of nucleic acid molecules comprising or consisting of:

[0087] In one particular embodiment of the invention, the nucleic acid molecule or combination of nucleic acid molecules encoding an antibody of the invention or an antigen-binding fragment thereof or a mimetic thereof further encodes a light chain constant domain, in particular comprising the nucleic acid sequence of SEQ ID NO: 96, and / or encodes a heavy chain constant domain, in particular comprising the nucleic acid sequence of SEQ ID NO: 98 or SEQ ID NO: 99.

[0088] Any nucleic acid molecule according to the invention can be inserted into an expression vector, such as a plasmid, suitable for expression of the encoded sequence in a host cell.

[0089] Compound combinations The present invention further relates to a combination of compounds comprising a humanized antibody or antigen-binding fragment thereof as a first therapeutic agent and at least one second therapeutic agent.

[0090] The first therapeutic agent is a humanized anti-CLEC-1A antibody or an antigen-binding fragment thereof or a mimetic thereof according to any embodiment disclosed herein. The at least one second therapeutic agent is a chemotherapeutic agent, a radiotherapeutic agent, an immunotherapeutic agent, in particular a tumor-targeting antibody or an antigen-binding fragment thereof, including anti-hCD20-hIgG1, anti-hEGFR-hIgG1, anti-hHER2-hIgG1, in particular a tumor-targeting monoclonal antibody or an antigen-binding fragment thereof, in particular a tumor-targeting monoclonal antibody or an antigen-binding fragment thereof that activates and / or enhances the phagocytic ability of macrophages, in particular alemtuzumab, atezolizumab, bevacizumab, anti-hEGFR-hIgG1 monoclonal tumor-targeting antibodies, such as cetuximab, herceptin, panitumumab, anti-hCD20-hIgG1 monoclonal tumor-targeting antibodies, such as , rituximab, anti-hHER2-hIgG1 monoclonal tumor-targeted trastuzumab, anti-PDL-1 antibodies, and anti-CD47 antibodies, or other antibodies or monoclonal antibodies selected from the group consisting of anti-PD1 antibodies, anti-CTLA4 antibodies, agonist anti-CD137 antibodies, anti-CD28 antibodies, anti-CD127 antibodies, anti-bcl2 antibodies, and anti-SIRPa antibodies; and / or chemotherapeutic agents, and / or cell therapy agents (e.g., CAR-T cells), and / or radiotherapy agents, in particular cytotoxic agents with anti-proliferative, pro-apoptotic, cell cycle arresting, and / or differentiation-inducing effects, in particular cytotoxic agents selected from the group consisting of cytotoxic antibodies, alkylating agents, anthracyclines, antimetabolites, anti-microtubule agents, topoisomerase inhibitors, alkaloids, bleomycin, anti-neoplastic agents, cyclophosphamide. In particular, said immunotherapeutic agents include anti-CD3 agents, particularly anti-CD3 antibodies, anti-PD1 agents (particularly anti-PD1 antibodies), particularly PD1 antagonists, particularly antagonistic anti-PD1 antibodies, anti-PDL1 agents (particularly anti-PDL1 antibodies), particularly PDL1 antagonists, particularly antagonistic anti-PDL1 antibodies, anti-CTLA4 agents (particularly anti-CTLA4 antibodies), particularly CTLA4 antagonists, particularly antagonistic anti-CTLA4 antibodies, CD137 agonists, particularly agonist anti-CD137 antibodies, anti-CLEC-1 agents (particularly anti-CLEC-1 antibodies), anti-VEGF agents, particularly anti-VEGF antibodies, anti-CD19 agents, In particular, selected from the group consisting of anti-CD19 antibodies, and anti-CD47 agents (particularly anti-CD47 antibodies), particularly CD47 antagonists, particularly anti-CD47 antagonist antibodies, anti-SIRPa agents (particularly anti-SIRPa antibodies), particularly anti-SIRPa antagonists, particularly anti-SIRPa antagonist antibodies, anti-CD28 agents (particularly anti-CD28 antibodies), particularly anti-CD28 antagonists, particularly anti-CD28 antagonist antibodies, anti-Bcl-2 agents (particularly venetoclax, also referred to as ABT199 or GDC-0199), tyrosine / kinase pathway inhibitors, such as venetoclax.

[0091] In one particular embodiment, the first therapeutic agent is a humanized anti-CLEC-1A antibody or an antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein, and the at least one second therapeutic agent is cetuximab.

[0092] In one particular embodiment, the first therapeutic agent is a humanized anti-CLEC-1A antibody or an antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein, and the at least one second therapeutic agent is rituximab.

[0093] In one particular embodiment, the first therapeutic agent is a humanized anti-CLEC-1A antibody or an antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein, and the at least one second therapeutic agent is trastuzumab.

[0094] Tumor-targeting antibodies can be defined as therapeutic monoclonal antibodies that recognize tumor-specific membrane proteins, block cell signaling, and induce tumor killing via an Fc-driven innate immune response.

[0095] In one particular embodiment of the invention, the first therapeutic agent is an antibody defined by its CDR domains disclosed herein, or an antigen-binding fragment thereof or a mimetic thereof, and the second therapeutic agent is rituximab, trastuzumab, cetuximab, or an antibody or monoclonal antibody selected from the group consisting of an anti-PD1 antibody, an anti-PDL-1 antibody, an anti-CD47 antibody, and an anti-SIRPa antibody.

[0096] In one particular embodiment, the anti-CLEC-1A antibody, or antigen-binding fragment thereof, or mimetic thereof, - specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 3 and a light chain variable domain comprising or consisting of SEQ ID NO: 4, in particular with an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 3 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33, and is an antagonist of human CLEC-1, in particular an antibody or an antigen-binding fragment or a mimetic thereof which enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 3 and a light chain variable domain comprising or consisting of SEQ ID NO: 4, in particular compared to an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 3 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 4 and SEQ ID NO: 33; or - specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 5 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular with an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 5 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, and is an antagonist of human CLEC-1, in particular an antibody or an antigen-binding fragment or a mimetic thereof which enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 5 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular compared to an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 5 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33; or - specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 6 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular with an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 6 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33, and is an antagonist of human CLEC-1, in particular an antibody or an antigen-binding fragment or a mimetic thereof which enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 6 and a light chain variable domain comprising or consisting of SEQ ID NO: 7, in particular compared to an antibody comprising a heavy chain domain comprising or consisting of SEQ ID NO: 6 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 7 and SEQ ID NO: 33; or - specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor), competes for binding to the human CLEC-1A receptor with an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 8 and a light chain variable domain comprising or consisting of SEQ ID NO: 9, in particular with an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 8 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100 or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 9 and SEQ ID NO: 33, and is an antagonist of human CLEC-1; an antibody or an antigen-binding fragment or a mimetic thereof which particularly enhances phagocytosis of tumor cells and / or secondary necrotic cells by myeloid cells, in particular by dendritic cells and / or macrophages, compared to a negative control, in particular compared to an antibody comprising or consisting of a heavy chain variable domain comprising or consisting of SEQ ID NO: 8 and a light chain variable domain comprising or consisting of SEQ ID NO: 9, in particular compared to an antibody comprising or consisting of a heavy chain domain comprising or consisting of SEQ ID NO: 8 and SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO: 101 and a light chain domain comprising or consisting of SEQ ID NO: 9 and SEQ ID NO: 33; and a second therapeutic agent selected from the list defined herein above or below.

[0097] The term "chemotherapeutic agent" includes chemical compounds that are effective in inhibiting tumor growth. Chemotherapeutic agents may be conventional cytotoxic agents, i.e., compounds that cause irreversible lethal damage following exposure by disrupting DNA replication, mitosis, etc. These agents may have antiproliferative, proapoptotic, cell cycle arresting, and differentiation-inducing effects. These agents are preferentially selected from the group consisting of alkylating agents (cisplatin, chlorambucil, procarbazine, carmustine), anthracyclines and other cytotoxic antibiotics, antimetabolites (i.e., methotrexate, cytarabine, gemcitabine), antimicrotubule agents (i.e., vinblastine, paclitaxel, docetaxel), topoisomerase inhibitors (i.e., etoposide, doxorubicin), alkaloids (i.e., vincristine, vinblastine, vinorelbine, camptothecin), or bleomycin (which inhibits the incorporation of thymidine into DNA chains).

[0098] The inventors demonstrate that the combinatorial use of CLEC-1A antagonists, in particular humanized CLEC-1A antagonist antibodies, in combination with other therapeutic agents, in particular rituximab, improves the phagocytic capacity of macrophages, in particular M1 macrophages, and therefore that anti-CLEC-1A antagonist compounds are suitable for improving the therapeutic effect of a second therapeutic agent administered simultaneously, separately or sequentially.

[0099] In one particular embodiment, the therapeutic agents may be administered simultaneously, separately or sequentially in the treatment of a disease.

[0100] The present invention further relates to a pharmaceutical composition comprising an anti-CLEC-1A humanized antibody or antigen-binding fragment thereof or mimetic thereof, or nucleic acid molecule or combination of nucleic acid molecules according to any embodiment disclosed herein as a first therapeutic agent, either alone or in combination with a second therapeutic agent disclosed herein, together with a pharmaceutically suitable vehicle that is pharmaceutically acceptable for formulation to be administered to a patient in need thereof, which may in particular be a sterile isotonic saline solution (such as mono- or di-sodium phosphate, sodium chloride, potassium chloride, calcium chloride, or magnesium chloride, or a mixture of such salts), or a dried composition, in particular a lyophilized composition, which can be constituted into an injectable solution upon addition of sterile water or sterile saline, as the case may be.

[0101] The present invention further relates to a pharmaceutical composition comprising a first therapeutic agent as defined herein, in particular a humanized anti-CLEC-1A antagonist antibody or its antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein, either alone or in combination with a second therapeutic agent, and / or together with a pharmaceutically suitable vehicle as defined herein, for simultaneous, separate, or sequential administration to a patient in need thereof, in particular in combination therapy with other treatments, including the use of medicines including chemotherapeutic agents, radiotherapeutic agents, immunotherapeutic agents (e.g., tumor-targeting monoclonal antibodies), cell therapy agents (e.g., CAR-T cells), immunosuppressants, proapoptotic agents, antibiotics, targeted cancer therapies, and / or probiotics. Radiation therapy may involve radiation or the concomitant administration of a radiopharmaceutical to the patient. The source of radiation may be either external or internal to the patient being treated (radiation treatment may be, for example, in the form of external beam radiation therapy (EBRT) or brachytherapy (BT)). Targeted cancer therapy is a drug or other substance that blocks the growth and spread of cancer by interfering with specific molecules ("molecular targets") involved in cancer growth, progression, and spread. Targeted cancer therapy may also be referred to as a "molecularly targeted drug," "molecularly targeted therapy," "precision medicine," or similar names. In some embodiments, the targeted therapy consists of administering a tyrosine kinase inhibitor to a subject. The term "tyrosine kinase inhibitor" refers to any of a variety of therapeutic agents or drugs that act as selective or non-selective inhibitors of receptor tyrosine kinases and / or non-receptor tyrosine kinases. Tyrosine kinase inhibitors and related compounds are well known in the art and are described in U.S. Patent Publication No. 2007 / 0254295, which is incorporated herein by reference in its entirety.

[0102] The present invention further relates to a method of treating cancer in a human subject in need thereof, comprising the step of administering to the subject a therapeutically effective amount of a first therapeutic agent as defined herein, in particular a humanized anti-CLEC-1A antagonist antibody or an antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein, wherein said first therapeutic agent is used in combination with a conventional treatment, in particular a conventional treatment for cancer. The present invention further relates to a humanized anti-CLEC-1A antagonist antibody or an antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein for use in the treatment of cancer in combination with a conventional treatment for cancer.

[0103] The term "standard or conventional treatment" as used herein relates to any treatment for cancer (drugs, radiation therapy, etc.) that is usually administered to subjects with cancer.

[0104] In particular, the humanized anti-CLEC-1A antagonist antibody or its antigen-binding fragment or mimetic thereof is used in combination with a chemotherapeutic agent, a radiotherapeutic agent, an immunotherapeutic agent (e.g., a tumor-targeted monoclonal antibody), a cellular therapy agent (e.g., CAR-T cells), an immunosuppressant, a proapoptotic agent, an antibiotic, a targeted cancer therapy, and / or a probiotic.

[0105] The present invention further relates to the use of the anti-CLEC1A antibodies and antigen-binding fragments and mimetics thereof disclosed herein for the treatment of cancer. The term "cancer" has its general meaning in the art and relates to a group of diseases involving abnormal cell growth that has the potential to invade or spread to other parts of the body. The term "cancer" further includes both primary and metastatic cancers. Examples of cancers that can be treated by the methods and compositions of the present invention include, but are not limited to, bladder, blood, bone, bone marrow, brain, breast, colon, esophageal, gastrointestinal, gingival, head, kidney, liver, lung, nasopharyngeal, cervical, ovarian, prostate, skin, stomach, testis, tongue, or uterine cancers. Furthermore, cancer specifically includes the following histological types: malignant neoplasm; carcinoma; undifferentiated carcinoma; giant cell carcinoma and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; malignant gastrinoma; cholangiocarcinoma; hepatocellular carcinoma; mixed hepatocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyps; familial polyposis coli adenocarcinoma; solid tumor; malignant carcinoid tumor; bronchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; eosinophilic carcinoma; eosinophilic adenocarcinoma; basophilic carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma. adenocarcinoma); non-encapsulated sclerosing carcinoma; adrenocortical carcinoma; endometrioid carcinoma; adnexal carcinoma; apocrine gland carcinoma; sebaceous gland carcinoma; ceruminous gland carcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; invasive ductal carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; Paget's disease of the breast; acinic cell carcinoma; adenosquamous carcinoma; adenocarcinoma with squamous metaplasia; malignant thymoma; malignant ovarian stromal tumor; malignant theca cell tumor; malignant granulosa cell tumor; and malignant neuroblastoma tumor;Sertoli cell carcinoma;malignant Leydig cell tumor;malignant lipocytoma;malignant paraganglioma;malignant extramammary paraganglioma;pheochromocytoma;angioangiosarcoma;malignant melanoma;amelanotic melanoma;superficial spreading melanoma;malignant melanoma in giant pigmented nevus;epithelioid cell melanoma;malignant blue nevus;sarcoma;fibrosarcoma;malignant fibrous histiocytoma;myxosarcoma;liposarcoma;leiomyosarcoma;rhabdomyosarcoma;embryonal rhabdomyosarcoma;alveolar rhabdomyosarcoma;stromal sarcoma;malignant mixed tumor;Mixed Müllerian tumor; Nephroblastoma; Hepatoblastoma; Carcinosarcoma; Malignant mesenchymoma; Malignant Brenner tumor; Malignant phyllodes tumor; Synovial sarcoma; Malignant mesothelioma; Dysgerminoma; Embryonic carcinoma; Malignant teratoma; Malignant ovarian goiter; Choriocarcinoma; Malignant mesonephroma; Angiosarcoma; Malignant hemangioendothelioma; Kaposi's sarcoma; Malignant hemangiopericytoma; Lymphangiosarcoma; Osteosarcoma; Parosteal osteosarcoma; Chondrosarcoma; Malignant chondroblastoma Cystoma;Mesenchymal chondrosarcoma;Giant cell tumor of bone;Ewing's sarcoma;Malignant odontogenic tumor;Ameloblastic odontosarcoma;Malignant ameloblastoma;Ameloblastic fibrosarcoma;Malignant pinealoma;Chordoma;Malignant glioma;Ependymoma;Astrocytoma;Protoplasmic astrocytoma;Fibrous astrocytoma;Astroblastoma;Glioblastoma;Oligodendroglioma;Oligodendroglioma;Primitive neuroectodermal The tumor may be, but is not limited to, cerebellar sarcoma, ganglionic blastoma, neuroblastoma, retinoblastoma, olfactory nerve tumor, malignant meningioma, neurofibrosarcoma, malignant neurilemmoma, malignant granular cell tumor, malignant lymphoma, Hodgkin's disease, Hodgkin's lymphoma, lateral granuloma, small lymphocytic lymphoma, diffuse large cell lymphoma, follicular lymphoma, mycosis fungoides, other specific types of non-Hodgkin's lymphoma, malignant histiocytosis, multiple myeloma, mast cell sarcoma, immunoproliferative small intestinal disease, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma cell leukemia, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myeloid sarcoma, and hairy cell leukemia.

[0106] In some embodiments, the subject has a cancer selected from the group consisting of bile duct cancer, bladder cancer, bone cancer, brain and central nervous system cancer, breast cancer, Castleman's disease, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gallbladder cancer, gastrointestinal carcinoid tumor, Hodgkin's disease, non-Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, pharyngeal and hypopharyngeal cancer, liver cancer, lung cancer, mesothelioma, plasmacytoma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, oral cavity and oropharyngeal cancer, ovarian cancer, pancreatic cancer, penile cancer, pituitary cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, vaginal cancer, vulvar cancer, and uterine cancer.

[0107] The present invention further relates to the use of the anti-CLEC1A antibodies and antigen-binding fragments and mimetics thereof disclosed herein for the treatment, including prophylactic treatment, of adverse conditions or diseases, particularly those involving the phagocytic capacity of myeloid cells, particularly dendritic cells and / or macrophages. In a particular embodiment, the disease or condition is selected from the group consisting of cancer, particularly those cancers listed herein above, in particular liquid cancers, solid cancers, lymphoma, colorectal cancer, mesothelioma, or liver cancer.

[0108] The present invention further relates to the use of the anti-CLEC1A antibodies and antigen-binding fragments and mimetics thereof disclosed herein, particularly for the treatment, including prophylactic treatment, of adverse conditions or diseases where stimulation of the phagocytic capacity of dendritic cells can ameliorate or treat the condition or disease. In a particular embodiment, the disease or condition is selected from the group consisting of cancer, particularly the cancers listed herein above, in particular liquid cancers, solid cancers, lymphoma, colorectal cancer, mesothelioma, or liver cancer.

[0109] The present invention further relates to the use of the anti-CLEC1A antibodies and antigen-binding fragments and mimetics thereof disclosed herein for the treatment, including prophylactic treatment, of any disease or condition susceptible to amelioration or prevention by increasing the phagocytic capacity of myeloid cells, particularly dendritic cells and / or macrophages, in particular where the disease or condition is selected from the group consisting of cancer, in particular any of the cancers listed herein above, in particular liquid cancers, solid cancers, lymphoma, colorectal cancer, mesothelioma, or liver cancer.

[0110] The present invention further relates to the use of the anti-CLEC1A antibodies and antigen-binding fragments and mimetics thereof disclosed herein for the treatment, including prophylactic treatment, of adverse conditions or diseases, particularly those involving T cells and involving T cell proliferation. In a particular embodiment, the disease or condition is selected from the group consisting of cancer, particularly those cancers listed herein above, in particular liquid cancers, solid cancers, lymphoma, colorectal cancer, mesothelioma, or liver cancer.

[0111] The present invention further relates to a method for increasing the phagocytic capacity of bone marrow cells, in particular dendritic cells and / or macrophages, comprising administering to a patient in need thereof an effective amount of an anti-CLEC-1A antibody or its antigen-binding fragment or mimetic thereof according to any embodiment disclosed herein; in particular, the antibody or its antigen-binding fragment or mimetic thereof is administered simultaneously, separately or sequentially with a conventional treatment or with at least one second therapeutic agent as defined herein.

[0112] The present invention further relates to an antibody or an antigen-binding fragment thereof or a mimetic thereof according to the invention, and / or a nucleic acid molecule or a combination of nucleic acid molecules according to the invention, and / or a combination of compounds and / or a pharmaceutical composition according to the invention for use as a medicament.

[0113] The present invention further relates to the use of a humanized anti-CLEC1A antibody or an antigen-binding fragment thereof or a mimetic thereof according to any embodiment disclosed herein, and / or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, and / or a combination of compounds and / or a pharmaceutical composition according to the invention, for the manufacture of a medicament. In particular, the present invention relates to the use of such an anti-CLEC-1A antibody or an antigen-binding fragment thereof or a mimetic thereof, and / or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, and / or a combination of compounds and / or a pharmaceutical composition according to the invention, for the manufacture of a medicament for the treatment and / or prevention of cancer, in particular the cancers listed herein above, in particular liquid cancers and solid cancers, in particular lymphoma, colorectal cancer, mesothelioma or liver cancer.

[0114] The present invention further relates to a method for treating or preventing a disease by administering a therapeutic amount of a humanized anti-CLEC1A antibody or antigen-binding fragment or mimetic thereof according to any of the definitions disclosed herein, and / or a nucleic acid molecule or combination of nucleic acid molecules according to the present invention, and / or a combination of compounds and / or a pharmaceutical composition according to the present invention to a patient in need thereof. In particular, the present invention relates to a method for treating or preventing cancer, in particular the cancers listed herein above, in particular liquid cancers and solid cancers, in particular lymphoma, colorectal cancer, mesothelioma, or liver cancer.

[0115] The present invention further relates to the use of the compounds, compositions and combinations of compounds as defined herein, in particular for the prevention or treatment of diseases or disorders. Accordingly, there is provided a humanized antibody or antigen-binding fragment thereof or a mimetic thereof according to the disclosure herein, or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, or a combination of compounds according to the invention, for use in the prevention and / or treatment of a disease or disorder, in particular a human disease or disorder, in which increased phagocytic capacity by myeloid cells, in particular dendritic cells and / or macrophages, ameliorates or prevents the disease or disorder.

[0116] Further provided is a humanized anti-CLEC1A antibody of the invention or an antigen-binding fragment or mimetic thereof, or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, or a combination of compounds according to the invention, for use in treating a disease or condition in which induction of phagocytosis in a patient ameliorates or prevents the disease or condition.

[0117] Further provided is a humanized anti-CLEC1A antibody of the invention or an antigen-binding fragment thereof or a mimetic thereof, or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, or a combination of compounds according to the invention, for use in treating patients with cancer, in particular liquid or solid cancer, in particular lymphoma, colorectal cancer, mesothelioma or liver cancer, inflammatory disease, chronic infection, or sepsis.

[0118] Further provided is a humanized antibody or an antigen-binding fragment thereof or a mimetic thereof of the invention, or a nucleic acid molecule or combination of nucleic acid molecules according to the invention, or a combination of compounds according to the invention, for use in combination therapy in which a first medicament comprising a chemotherapeutic agent, a radiotherapeutic agent, an immunotherapeutic agent (e.g., a tumor-targeted monoclonal antibody), a cellular therapy agent (e.g., CAR-T cells), an immunosuppressant, a proapoptotic agent, an antibiotic, a targeted cancer therapy, and / or a probiotic is administered to a patient in need thereof, particularly for simultaneous, separate, or sequential administration.

[0119] [Table 1]

[0120] Any combination of VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 is contemplated in the present invention.

[0121] [Table 2]

[0122] The variable heavy chains listed herein correspond to those present in antibodies 11H11, 14H9, and 6C5 used in the examples of the present invention.

[0123] [Table 3]

[0124] The variable light chains listed herein correspond to those present in the antibodies 11H11, 14H9, and 6C5 used in the examples of the present invention.

[0125] Any combination of a heavy chain variable domain selected in Table 2 with a light chain variable domain selected in Table 3 is contemplated in the present invention.

[0126] The following figures and examples are presented so as to provide those of ordinary skill in the art with a complete disclosure and description of the manner of making and using the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below are all or the only experiments that will be performed. While the invention has been described in connection with specific embodiments thereof, those skilled in the art will recognize that various modifications can be made and equivalents can be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps to the objective, spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto. [Brief explanation of the drawings]

[0127] [Figure 1-1] Phagocytosis assays of tumor cells (non-Hodgkin's lymphoma cells (Raji cells) (A); colon cancer cells (DLD-1) (B); breast cancer cells (SK-BR3) (C); non-Hodgkin's lymphoma cells (Raji cells) (D); and lung cancer cells (NSCLC) (E) are shown in A, B, and C in the presence of humanized antibodies according to the invention (11H11, 14H9, and 6C5) compared with prior art anti-CLEC-1A antibodies (αCLEC-1 Ctrl mAb-D and E) corresponding to the anti-CLEC-1A antibodies disclosed in WO2018073440 and Robles et al. (Blood Advances, 2017). The isotype control was an irrelevant humanized antibody. The phagocytosis rate was determined by normalizing the frequency of Clec-1-blocked TGFb-DCs that phagocytosed tumor cells to PBS or the control antibody by the isotype of the mAb used. [Figure 1-2]Phagocytosis assays of tumor cells (non-Hodgkin's lymphoma cells (Raji cells) (A); colon cancer cells (DLD-1) (B); breast cancer cells (SK-BR3) (C); non-Hodgkin's lymphoma cells (Raji cells) (D); and lung cancer cells (NSCLC) (E) are shown in A, B, and C in the presence of humanized antibodies according to the invention (11H11, 14H9, and 6C5) compared with prior art anti-CLEC-1A antibodies (αCLEC-1 Ctrl mAb-D and E) corresponding to the anti-CLEC-1A antibodies disclosed in WO2018073440 and Robles et al. (Blood Advances, 2017). The isotype control was an irrelevant humanized antibody. The phagocytosis rate was determined by normalizing the frequency of Clec-1-blocked TGFb-DCs that phagocytosed tumor cells to PBS or the control antibody by the isotype of the mAb used. [Figure 2] Phagocytosis assays of tumor cells (non-Hodgkin's lymphoma cells (Raji cells) (A); colon cancer cells (DLD-1) (B); breast cancer cells (SK-BR3) (C); and non-Hodgkin's lymphoma cells (Raji cells) (D) are shown in the presence of combinations of humanized antibodies according to the invention (11H11, 14H9, and 6C5) with 1 ng / ml rituximab (A); cetuximab (B), and trastuzumab (C), compared to a prior art anti-CLEC-1A antibody (αCLEC-1 Ctrl mA corresponding to the anti-CLEC-1A antibody disclosed in WO2018073440 and Robles et al. (Blood Advances, 2017)) in combination with 1 ng / ml rituximab in D. The isotype control is an irrelevant humanized antibody. The rate of phagocytosis was determined by normalizing the frequency of Clec-1-blocked TGFb-DCs that phagocytosed tumor cells to control antibodies with PBS or an isotype control for the mAb used. [Figure 3]This figure shows the effect of the anti-CLEC1 antibody of the present invention on a mouse model of liver cancer (HCC model). The antitumor effect of ip administration of anti-CLEC1 antibodies (humanized 6C5 or humanized 14H9, 3 mg / kg) twice a week for 3 weeks in an orthotopic mouse hepatoma model (2.5 × 10 Hepa1.6 cells were injected via the portal vein on day 0). An isotype control antibody was used in the control group (3 mg / kg). [Figure 4]

[0039] Figure 1 shows the effect of the anti-CLEC1 antibody of the present invention on a mouse model of colorectal cancer (CRC model). Antitumor effect of ip administration of an anti-CLEC1 antibody (humanized 6C5, 3 mg / kg) twice weekly for 3 weeks in combination with chemotherapy (100 mg / kg cyclophosphamide once tumors reached 50-100 mm) in a syngeneic mouse model of colorectal cancer (0.5 x 10 MC38 cells injected subcutaneously on day 0). An isotype control antibody was used in the control group (3 mg / kg). [Figure 5] Figure 1 shows antagonist activity test of humanized anti-CLEC1 antibodies against the interaction of Fc-CLEC1-permeable Raji by FACS. Different anti-CLEC1 antibodies were tested in a dose-response manner: 6C5, 14H9, and 1H11 (humanized anti-CLEC-1A antibodies of the invention), as well as an isotype control as a negative control. The curve represents the percentage binding of Fc-CLEC1-A488 at 10 nM to Raji cells after competition with the anti-CLEC1 antibodies. Cytometric evaluation of permeable Raji using A488-labeled FcCLEC at a constant concentration (10 nM), as well as humanized 14H9 (black triangles), humanized 11H11 (◆), humanized 6C5 (□), and an isotype control (+). Revelation was performed using a CytoFlex cytometer, and values ​​correspond to the percentage (%) of stained cells. [Figure 6]Figure 1 shows antagonist activity testing of anti-CLEC1 antibodies (including the humanized anti-CLEC-1A antibodies 14H9, 11H11, and 6C5 of the invention) on the Fc-CLEC1-permeable NALM6 cell line. The curve shows the percentage of binding of Fc-CLEC1-A488 at 10 nM to PBMCs after competition with anti-hCLEC1 antibodies. Cytometric evaluation on permeabilized NALM6 using A488-labeled FcCLEC at a constant concentration (100 nM), as well as humanized 14H9 (black triangles), humanized 11H11 (◆), humanized 6C5 (□), and an isotype control (+). Revelation was performed using a CytoFlex cytometer, and values ​​correspond to the percentage (%) of stained cells. [Figure 7] Figure 1 shows antagonist activity testing of anti-CLEC1 antibodies (including the humanized anti-CLEC-1A antibodies 14H9, 11H11, and 6C5 of the invention) on the Fc-CLEC1 native non-permeabilized NALM6 cell line. The curve shows the percentage of binding of Fc-CLEC1-A488 at 100 nM to NALM6 after competition with anti-hCLEC1 antibodies. Cytometric evaluation on non-permeabilized NALM6 using A488-labeled FcCLEC at a constant concentration (100 nM), as well as humanized 14H9 (black triangles), humanized 11H11 (◆), humanized 6C5 (□), and an isotype control (+). Revelation was performed using a CytoFlex cytometer, and values ​​correspond to the percentage (%) of stained cells. [Figure 8] Figure 1 shows antagonist activity tests of humanized and chimeric anti-CLEC1 antibodies (6C5 antibody (A); 1H11 antibody (B); and 14H9 antibody (C)) against the Hepa1.6 cell line. Table (D) shows the EC50 of each humanized and chimeric antibody. Curves (A-C) represent the percentage of Fc-CLEC-1-positive viable cells according to the concentration of anti-CLEC-1 chimeric (open circles) or humanized (filled circles) mAb, normalized to the isotype control antibody condition. EC50 relates to the concentration required for each humanized and chimeric anti-CLEC1A antibody to achieve 50% of the maximum signal in the assay. [Figure 9]Figure 1 shows antagonist activity tests of humanized and chimeric anti-CLEC1 antibodies (6C5 antibody (A) and 1H11 antibody (B)) against the MC38 cell line. Table (C) shows the EC50 of each humanized and chimeric antibody. Curves (A-C) represent the percentage of Fc-CLEC-1-positive viable cells according to the concentration of anti-CLEC-1 chimeric (open circles) or humanized (filled circles) mAb, normalized to the isotype control antibody condition. EC50 relates to the concentration required for each humanized and chimeric anti-CLEC1A antibody to achieve 50% of the maximum signal in the assay. [Figure 10] Figure 1 shows the productivity of humanized anti-CLEC-1A antibodies 6C5, 14H9, and 11H11 of the invention in HEK cells (A) and CHO cells (B). Table (C) shows the recovered antibody yield compared to the control anti-CLEC-1A antibody production in the bottom row. [Figure 11] 1 shows the binding affinity (KD), affinity constant (ka), and dissociation constant (kd) of the humanized antibodies of the present invention to human CLEC-A-his recombinant protein, as measured by Blitz. [Figure 12] Figure 1 shows binding studies of different humanized anti-CLEC1 antibodies of the invention in dose response to immobilized CLEC-1A-his recombinant protein (A) and Fc-CLEC-1A (B) by ELISA. [Figure 13] 1 shows a binding test of the humanized CLEC1 antibody of the present invention on the human U266 cell line by flow cytometry (FACS) by ELISA. [Figure 14-1] Binding studies of different humanized CLEC1 antibodies of the invention and their mutated versions are shown. [Figure 14-2] Binding studies of different humanized CLEC1 antibodies of the invention and their mutated versions are shown. DETAILED DESCRIPTION OF THE INVENTION

[0128] Materials and Methods Preparation and characterization of humanized anti-clec antibodies For humanized anti-clec, the variable sequences of the heavy chain (VH) of the anti-clec antibody were cloned by EcoRV into the pcDNA3.4 human G4m expression plasmid (OSE immunotherapy plasmid) containing the CH1-hinge-CH2-CH3 domains of hIgG4 mutated with S228P to stabilize the hinge region. The variable sequences of the light chain (VL) of the anti-clec antibody were cloned by BsiWI into the pcDNA3.4 CLIg-hkappa expression plasmid (OSE immunotherapy plasmid) containing human CL kappa. HEK freestyle cells or CHO mammalian cells were cotransfected with the same plasmids containing VH-hFcG4m and VL-CLhk using the lipofectamine or polyethyleneimine method. After 6–7 days of incubation, supernatants were collected and purified by affinity chromatography with Protein A (HiTrap, GeHealthcare) using 0.1 M citrate pH 3 elution buffer. Purified antibodies were concentrated by dialysis against PBS, 100 mM arginine / L-glutamate. Antibodies were quantified by UV at 280 nm and tested in several assays: ELISA and activity assays against Clec-his in Blitz (ForteBio), activity assays on U266 cells (where Clec is present on the cell surface), and antagonist assays using permeabilized cell lines.

[0129] Preparation and characterization of humanized anti-CLEC1 antibody variants Humanized anti-CLEC1 antibody variants (11H11m, 6C5m1, 6C5m2, and 14H9m) were generated by PCR using primers containing substituted nucleotides with the Q5® Site-Directed Mutagenesis Kit (NEBiolabs), transformed into Top10 chemically competent cells, and then plasmid purified. CHO mammalian cells were cotransfected with the same plasmids containing VH-hFcG4m and VL-CLhk by the polyethylenimine method. After 6–7 days of incubation, supernatants were collected and quantified by sandwich ELISA and CLEC1 binding assays.

[0130] ELISA Quantitative Sandwich Assay For quantitative ELISA assays, donkey anti-human IgG, Fc-specific antibody (Jackson Immunoresearch, no. 709-005-098) was immobilized on plastic at 1.3 μg / ml, and binding was measured by adding antibody-containing supernatant or purified antibody. After incubation and washing, mouse anti-human kappa antibody (OSE immunotherapeutics; no. NaM76-5F3) was added, followed by peroxidase-labeled donkey anti-mouse IgG (Jackson immunoresearch, no. 715-036-151) and developing by conventional methods.

[0131] ELISA activity assay of human anti-CLEC-1A with CLEC1-His coating For activity ELISA assays, recombinant hCLEC-His (R et D systems; ref. 1704-CL) was immobilized on plastic at 2 μg / ml and binding was measured by adding antibody-containing supernatant or purified antibody. After incubation and washing, peroxidase-labeled donkey anti-human IgG (Jackson immunoresearch, ref. 709-035-149) was added and developed by conventional methods.

[0132] ELISA activity assay of human anti-CLEC-1A with Fc-CLEC1 coating For activity ELISA assays, recombinant hFc-CLEC (OSE Immunotherapeutics; Nantes) were immobilized on plastic at 2 μg / ml, and antibody-containing supernatants or purified antibodies were added to measure binding. After incubation and washing, mouse anti-human kappa antibodies (OSE immunotherapeutics; ref. NaM76-5F3) were added, followed by peroxidase-labeled donkey anti-mouse IgG (Jackson immunoresearch, ref. 715-036-151), and development was performed using conventional methods.

[0133] Anti-Clec binding assay on U266 cells by cytofluorometry To measure anti-clec binding on U266 CLEC1+ cells, a 1 / 50 diluted human Fc receptor binding inhibitor (BD Pharmingen, USA; Ref. No. 564220) was first added for 30 min at room temperature to block human Fc receptors on U266 cells and reduce background. The antibody was then incubated for 30 min at 4°C, washed, and then stained with PE-labeled anti-human IgG Fc (Biolegend, USA; Ref. No. 409303) for 15 min at 4°C. Samples were analyzed using a cytoflex (Beckman Coulter).

[0134] Affinity analysis of anti-clec antibodies to human CLEC-His recombinant proteins by Blitz Clec-His recombinant protein (R et D systems; reference number 1704-CL) was immobilized on a NINTA biosensor at 10 μg / ml, and the indicated antibodies were added at 20 μg / ml. After an association time (k) of 120 s followed by a dissociation time (kd) of 120 s, values ​​were extracted to determine the affinity constant (KD).

[0135] Antagonist activity by flow cytometry For the competition assay, Fc-Clec-1 (Ose Immunotherapeutics, Nantes, France) was conjugated with Alexa Fluor 488 (Alexa Fluor® 488 Microscale Protein Labeling Kit, number A30006, Fisher Scientific, Illkirch, France). Permeabilized (reference number 554714, CytoFix / Cytoperm kit, BD Biosciences, France) and Fc-blocked (reference number 564220, BD Biosciences) Burkitt's lymphoma Raji cells express Clec-1 ligand, which can be detected after incubation with 10 nM Alexa488-labeled Fc-Clec. To measure competition, purified anti-Clec antibodies at different concentrations were preincubated with Alexa488-labeled Fc-Clec (constant 10 nM) for 15 min at room temperature. The preincubated mixture was then incubated on permeabilized and Fc-blocked Raji cells for 30 min on ice. Binding on the cells was then fixed with 2% PFA in cold PBS for 10 min on ice and analyzed with a CytoFlex cytofluorometer (Beckman Coulter France, Villepinte). Competition was also measured on NALM6 cells, which can express the ligand intracellularly or on the extracellular surface. Purified anti-Clec antibodies were preincubated with 100 nM Alexa488-labeled Fc-Clec at different concentrations for 15 minutes at room temperature. The preincubated mixture was then incubated on Fc-blocked NALM6 cells, with or without prepermeabilization, for 30 minutes on ice. Binding on the cells was then fixed with 2% PFA in cold PBS for 10 minutes on ice and analyzed using a CytoFlex cytofluorometer (Beckman Coulter France, Villepinte).

[0136] Phagocytosis assay Monocytes were isolated by magnetic sorting from hematoadsorption of healthy subjects using a Miltenyi classical monocyte isolation kit. Immature dendritic cells (iDCs) were then generated by culturing monocytes with 50 ng / mL human recombinant GM-CSF (CellGenix) and 20 ng / mL human recombinant IL-4 (CellGenix) for 6–7 days. iDCs were polarized into tolerogenic DCs with 50 ng / mL human recombinant TGFb (PeproTech) for 2 days, resulting in Clec-1 overexpression by TGFb-DCs. Antibodies were added at 10 μg / mL during polarization. TGFβ-DCs were cultured with non-Hodgkin's lymphoma (Raji) cells at a 1:1 ratio with 10 ng / mL of anti-CD20 mAb (rituximab) to generate an "eat-me" signal; naked NSCLC cells (A549) were cultured with TGFβ-DCs for 5 days. Phagocytosis was analyzed by flow cytometry and normalized to the control antibody condition for each donor. Macrophages (MΦ) were generated from monocytes using M-CSF (100 ng / mL) for 5 days. MΦ were incubated with either non-Hodgkin's lymphoma (Raji; CD20+), colon cancer (DLD-1; EGFR2+), or breast cancer (SK-BR3; Her2+) cells at a 1:2 ratio with or without 10 ng / mL of anti-CD20 mAb (rituximab), anti-EGFR mAb (cetuximab), or anti-Her2 mAb (trastuzumab), resulting in an "Eat-me" signal for 2 hours. Phagocytosis analysis was performed by flow cytometry, and the percentage of phagocytosis was calculated as the percentage of CPDe670+ cells among total CPDe450+ cells. Results were expressed as the percentage of MΦs that phagocytosed Raji cells multiplied by the median fluorescence intensity of phagocytic cells and expressed according to the concentration of rituximab. For macroscopic assays, macrophages were generated as described above. MΦs were preincubated with anti-CLEC1 chimeric mAb for 2 hours, followed by the addition of non-Hodgkin's lymphoma (Raji; CD20+) and anti-CD20 mAb (rituximab) at 10 ng / mL each for 4 hours to generate an "Eat-me" signal. Phagocytosis analysis was performed by microscopy, and the percentage of phagocytosis was calculated as the percentage of pHrodo (pHrodo-SE, Thermofisher)-positive Raji among total macrophages. To characterize cells in the phagocytosis assay, tumor cell lines, Raji (B lymphoma) CSCLC cells, colorectal cancer cells, and breast cancer cells, Huh7 (liver cancer), were stained with fluorescent dyes. Briefly, tumor cells were incubated with Cell Proliferation Dye eFluor 670 for 15 min and washed before UV treatment according to the manufacturer's instructions (Life Technologies). Cells were then irradiated with 150 mJ / cm2 of UV light. 2 The cells are treated with 1000 μL of UV light and incubated overnight to induce an apoptotic program that results in Clec-1 ligand expression. TGFb-DCs and tumor cell lines were collected, counted, and incubated for 5 hours at a ratio of 2 DCs to 1 tumor cell, with antibody added at 10 μg / mL during the process. Phagocytosis was assessed by flow cytometry on CPD-eFluor670-positive TGFb-DCs.

[0137] In the examples of the present invention, unless otherwise stated, the anti-CLEC-1A control antibody is an in-house antibody that does not have antagonistic properties. [Example]

[0138] Example 1 Biological activity of the humanized anti-hCLEC1A antagonist antibodies of the present invention and anti-hCLEC1A antagonist antibodies disclosed in the prior art on tumor phagocytosis by dendritic cells - Figure 1 method a) Generation of monocyte-derived dendritic cells (DCs) polarized with TGFb recombinant protein Monocytes were isolated by magnetic sorting from hematoadsorption from healthy subjects using a Miltenyi classical monocyte isolation kit. Immature dendritic cells (iDCs) were then generated by culturing monocytes with 50 ng / mL human recombinant GM-CSF (CellGenix) and 20 ng / mL human recombinant IL-4 (CellGenix) for 6–7 days. iDCs were polarized into tolerogenic DCs with 50 ng / mL human recombinant TGFb (PeproTech) for 2 days, resulting in Clec-1 overexpression by TGFb-DCs. Antibodies were added at 10 μg / mL during polarization. b) Generation of UV-treated apoptotic tumor cell lines To characterize cells in the phagocytosis assay, tumor cell lines representing non-Hodgkin's B lymphoma (Raji cells - Figures 1A and 1D), colon cancer (DLD-1 model - Figure 1B), breast cancer (SK-BR3 cell line - Figure 1C), and lung cancer (NSCLC cells - Figure 1E) were stained with fluorescent dyes. Briefly, tumor cells were incubated with Cell Proliferation Dye eFluor 670 for 15 min and washed before UV treatment according to the manufacturer's instructions (Life Technologies). Cells were then irradiated with 150 mJ / cm2 of light. 2 The cells are treated with 1000 μL of UV light and incubated overnight to induce an apoptotic program that results in Clec-1 ligand expression. c) Phagocytosis assay TGFb-DCs and tumor cell lines were harvested, counted, and incubated for 5 hours at a ratio of 2 DCs to 1 tumor cell, with the antibody added at 10 μg / mL during the incubation process. Phagocytosis was assessed by flow cytometry on CPD-eFluor670-positive TGFb-DCs. After staining with eFluor450 cell proliferation dye (Life Technologies), humanized anti-Clec-1 mAb was incubated with human macrophages (monocyte-derived macrophages polarized with IFNγ) at 37°C for 1 hour. Results. Figure 1 shows the phagocytosis of UV-treated tumor cells by TGFb-DCs, normalized to the control condition. In three different cancer models: lymphoma (Figure 1A), carcinoma (Figure 1B), and breast cancer (Figure 1C), the antagonist humanized 14H9, 6C5, and 11H11 antibodies of the present invention increased tumor cell phagocytosis, whereas prior art control antibodies (disclosed in WO2018073440 and Robles et al., Blood Advances, 2017) did not induce any significant changes in the ability of DCs to phagocytose tumor cells in lymphoma (Figure 1D) or lung cancer (Figure 1E). Phagocytosis of UV-treated tumor cells by TGFb-DCs was improved in the presence of antibodies of the present invention compared to negative controls, normalized to the control condition, in all cancer models. This example demonstrates the ability of the humanized antibodies of the present invention to increase tumor cell phagocytosis by dendritic cells, in contrast to prior art antibodies (disclosed in WO 2018073440 and Robles et al., Blood Advances, 2017). It is noted that the humanized anti-CLEC-1A antibodies 6C5 and 14H9 were IgG1 antibodies (hum 6C5-m, hum 14H9-m) with constant chain domains corresponding to IgG1N297A of SEQ ID NO: 97. The humanized 11H11 antibody was an IgG4 antibody (S228P) (hum 11H11). According to the illustrated results, tumor cell phagocytosis appears to be significantly greater when the humanized antibody is either 6C5 or 14H9. Providing IgG1 humanized anti-CLEC-1A antibodies may be interesting compared to providing their IgG4 equivalents.

[0139] Example 2 Biological activity of the anti-hCLEC-1A antagonist antibodies of the present invention or of the prior art in combination with tumor-targeting antibodies: rituximab, cetuximab, or trastuzumab - Figure 2 method After staining with eFluor450 cell proliferation dye (Life Technologies), humanized anti-Clec-1 mAb was incubated at 10 μg / mL with human macrophages (monocyte-derived macrophages polarized with IFNγ) at 37°C for 1 h. After staining with eFluor 647 cell proliferation dye (Life Technologies), human Raji B lymphoma, DLD-1 colon carcinoma, and SK-BR3 breast cancer cell lines were incubated with medium or 1 ng / mL of anti-tumor-associated antigen (TAA) (rituximab anti-CD20 for Raji, cetuximab anti-EGFR for DLD-1, and trastuzumab anti-Her2 for SK-BR3) for 1 h at 37°C. Anti-CLEC-1-treated macrophages were incubated with tumor cell lines with or without anti-TAA opsonization for 1 h at 37°C for phagocytosis assays. Phagocytosis analysis was performed by flow cytometry, and the percentage of phagocytosis was calculated as the percentage of CPDe670+ cells among total CPDe450+ cells. Results were expressed as the percentage of M1s that phagocytosed Raji cells multiplied by the median fluorescence intensity of phagocytic cells and expressed according to the rituximab concentration. Results: Phagocytosis assays show that M1 macrophages are able to phagocytose Raji cells in the presence of a combination of rituximab and an anti-CLEC-1A antibody of the present invention (Figure 2A), compared to prior art antibodies disclosed in WO2018073440 and Robles et al. (Blood Advances, 2017) (see Figure 2D). The same results are demonstrated for two other cancer models: administration of a combination of cetuximab and an anti-CLEC-1A antibody of the present invention increases phagocytosis of colon cancer tumor cells by macrophages (Figure 2B); and administration of a combination of trastuzumab and an anti-CLEC-1A antibody of the present invention increases phagocytosis of breast cancer tumor cells by macrophages (Figure 2C). The combination of an anti-CLEC-1A antibody of the present invention with a second anti-tumor antibody improves the phagocytic ability of M1 macrophages. Thus, it is demonstrated that the use of the anti-CLEC-1A antagonist antibodies of the present invention improves the therapeutic efficacy of tumor-targeting antibodies. This example demonstrates the ability of antibodies of the invention in combination with tumor-targeting antibodies to increase phagocytosis of tumor cells by macrophages, contrary to prior art antibodies (disclosed in WO2018073440 and Robles et al. (Blood advances, 2017)).

[0140] Example 3 Antitumor effect on overall survival in a mouse liver cancer tumor model - Figure 3 method Mice were anesthetized with an air / isoflurane mixture. After laparotomy, tumor-bearing Hepa1.6 cells were injected via the portal vein in PBS (2.5 × 10 cells / 100 μL). Treatment began 4 days after tumor injection. Anti-CLEC1 antibodies and isotype controls were injected at 3 mg / kg twice a week for 3 weeks. Overall survival was tracked, and the percentage of survival in each condition is reported in Figure 3. Results: As shown in Figure 3, animals treated with the humanized anti-CLEC1 antibodies of the invention exhibited extended survival rates for both anti-CLEC1 therapies (mice treated with control antibodies all died after 25 days, whereas mice treated with the anti-CLEC1 antibodies of the invention, 6C5 or 14H9, survived at least 2 weeks longer). These results demonstrate the unexpected efficacy of a therapeutic monotherapy (humanized anti-CLEC1 antibodies of the invention) against HCC tumor models.

[0141] Example 4 Antitumor effects on tumorigenesis in a mouse colorectal cancer tumor model treated with combination therapy (anti-CLEC1 antibody and chemotherapy) - Figure 4 method Mice were anesthetized with an air / isoflurane mixture. Tumor-bearing MC38 cells were injected subcutaneously in PBS (0.5 × 10 cells / 100 μL). Treatment began 4 days after tumor injection. The anti-CLEC1 antibody of the present invention, 6C5, and an isotype control were injected at 3 mg / kg twice weekly for 3 weeks. Chemotherapy was administered intraperitoneally once at 100 mg / kg in PBS when tumors reached 50-100 mm. Tumor development was assessed by measuring tumor length and width; tumor development was determined from baseline tumor measurements for each condition and is reported in Figure 4. Results: As shown in Figure 4, animals treated with the humanized anti-CLEC1 antibody of the present invention and chemotherapy showed superior response rates compared to chemotherapy alone. Measurement of the combined tumor volume in mice treated with anti-CLEC1 antibody and chemotherapy did not increase as much as in mice treated with chemotherapy alone after 40 days (600 mm 3 1500mm 3 Furthermore, 4 mice (out of 10) treated with the humanized anti-CLEC1 antibody 6C5 of the invention and chemotherapy survived the experiment, whereas all mice treated with chemotherapy alone died. This result demonstrates the unexpected efficacy of the therapeutic combination (humanized anti-CLEC1 antibody of the invention plus chemotherapy) against a CRC tumor model.

[0142] Example 5 Competition study between CLEC1 ligand and humanized anti-hCLEC1 antibody using antagonist assay - Figures 5 to 9 Methods. To measure competition for permeabilized Raji expressing the CLEC1 ligand, Fc-CLEC1-labeled A488, which specifically binds to permeabilized Raji (CytoFix / cytoperm kit, BD Biosciences), was used. To measure competition, 10 nM of Fc-CLEC1-labeled A488 was mixed with different concentrations of humanized anti-hCLEC1 for 15 minutes at room temperature and then added to the cells for 30 minutes at 4°C. After incubation and washing, 2% PFA was added to the wells to fix the cells for 10 minutes at 4°C. Cells were then analyzed on a CytoFlex (Beckman) cytofluorometer to detect inhibition of Fc-CLEC1 labeling. Measurement of competition for NALM6 cells. 100 nM of Fc-CLEC1-labeled A488 was mixed with different concentrations of humanized anti-hCLEC1 for 15 minutes at room temperature and then added to the cells for 30 minutes at 4°C. After incubation and washing, 2% PFA was added to the wells to fix the cells for 10 minutes at 4°C and analyzed on a CytoFlex (Beckman) cytofluorometer to detect inhibition of Fc-CLEC1 labeling. Mouse hepatoma Hepa1.6 or colorectal carcinoma MC38 cell lines were also used in the antagonist assay. Chimeric or humanized anti-Clec-1 mAbs (6C5, 11H11, 14H9) were also incubated at different concentrations (60 μg / mL to 0.08 μg / mL) with 10 μg / mL of human Fc-CLEC-1 recombinant protein conjugated with A488 fluorochrome for 30 minutes on ice. Mouse hepatoma Hepa1.6 or colorectal carcinoma MC38 cell lines were first stained with viability markers and then with A488 fluorochrome-conjugated recombinant human Fc-CLEC-1 protein preincubated with antagonist anti-CLEC-1 mAbs for 30 minutes on ice. Finally, cells were fixed with 1% PFA solution and read on a cytometer. The graph shows the percentage of Fc-CLEC-1 positive viable cells according to the concentration of anti-CLEC-1 chimeric (open circles) or humanized (filled circles) mAb, normalized to the hIgG4 control condition. Results: Figures 5-9 illustrate the antagonist activity of humanized anti-hCLEC1 antibodies of the present invention compared to an isotype control or an in-house chimeric anti-CLEC1 control (control + anti-Clec1). Fc-CLEC1 was able to specifically bind to permeabilized Raji, permeabilized NALM6 cells, and native NALM6 cells at 10 nM and 100 nM, respectively. The three antibodies tested dose-dependently blocked the interaction of Fc-CLEC with its ligands on permeabilized Raji, permeabilized NALM6 cells, and native NALM6 cells, compared to the isotype control, which did not inhibit Fc-CLEC binding on those cells. The IC50 values ​​and inhibition profile curves were similar among the three antibodies (see Figures 5-7). In complementary assays, we assessed the ability of the humanized antibodies of the present invention to antagonize human CLEC-1A to compare the results achieved with their chimeric counterparts. As shown in Figures 8 and 9, all three humanized antibodies, 11H11, 14H9, and 6C5, antagonized Fc-CLEC1A binding to the mouse hepatoma Hepa1.6 cell line or the colorectal carcinoma MC38 cell line (Figures 8A-8C and 9A-9B). At the lowest concentrations, the humanized antibodies were better antagonists than their chimeric counterparts. As shown in the tables (Figures 8D and 9C), the EC50 values ​​of the humanized antibodies were at most half that of their chimeric counterparts, often ranging between 5- and 3-fold lower. Thus, all tested humanized antibodies of the invention are able to interfere with the binding between CLEC-1A and cells that normally bind CLEC-1A, thereby demonstrating that they can antagonize the binding between CLEC-1A and one of its ligands. Thus, this example demonstrates that the antibodies of the invention are antagonists of human CLEC-1. Furthermore, the humanized antibodies of the invention appear to be better antagonists of CLEC-1A than their chimeric counterparts, especially at low concentrations.

[0143] Example 6 Production of humanized anti-CLEC1 antibodies - Figure 10 Mammalian HEK and CHO cells were cotransfected with plasmids containing VH-hFcG4m or VH-hFcG1N297A, respectively, along with a plasmid containing VL-CLkappa, using the lipofectamine method or polyethyleneimine (PEI). After 5–6 days of incubation, the supernatant was purified by affinity chromatography on Protein A (HiTrap, GeHealthcare) with 0.1 M citrate pH 3 elution buffer. The purified antibody was concentrated by dialysis against PBS, 100 mM arginine / L-glutamate. Antibody was quantified by UV (A280 nm), and the yield corresponds to the amount of purified antibody per liter of harvested culture supernatant. As shown in Figures 10A and 10B, the antibodies of the present invention were well expressed with different productivity (signal peptide used: IgKleader). As shown in the table (Figure 10C), the humanized antibodies had high production yields in HEK cells and CHO cells. This example demonstrates that the antibodies of the present invention can be efficiently produced in a recombinant production system.

[0144] Example 7 CLEC1 binding assay of humanized anti-hCLEC1 antibodies by ELISA - Figures 11 to 14 Methods: Affinity analysis of anti-CLEC1 antibodies against human CLEC-His recombinant protein was performed by Blitz. CLEC1-His recombinant protein was immobilized on a NINTA biosensor, and the indicated antibodies were added at 20 μg / ml. After an association time (ka) of 120 seconds followed by a dissociation time (kd) of 120 seconds, values ​​were extracted to determine the affinity constant (KD) (Figure 11). The binding activity of the anti-hCLEC1 antibodies was further evaluated by ELISA (enzyme-linked immunosorbent assay). For the ELISA assay, recombinant hCLEC1-His (R&D systems, reference number 1704-CL) was immobilized on plastic at 2 μg / ml in carbonate buffer (pH 9.2), and purified antibodies were added at different concentrations to measure binding. After incubation and washing, development was performed using peroxidase-conjugated donkey anti-human antibodies and colorimetry at 450 nm using TMB substrate (Jackson Immunoresearch; reference number 715-036-151) (Figure 12A). A second ELISA assay was performed as described above by immobilizing Fc-CLEC1 (OSE Immunotherapeutics) at 2 μg / ml. As previously described, ELISA was performed by immobilizing mouse Fc-CLEC1 (OSE Immunotherapeutics) at 2 μg / ml in carbonate buffer instead of His-Clec. Purified antibodies were added at different concentrations to measure binding. After incubation and washing, color development was performed using mouse anti-human kappa antibodies plus peroxidase-labeled donkey anti-mouse antibodies, and color development was measured at 450 nm using TMB substrate. ED50 is the concentration of the indicated antibody required to achieve 50% of the signal in the assay (Figure 12B). The control antibody was an isotype control. A third binding study was performed by cytofluorometry on the human U266 cell line CLEC1+ with humanized 14H9, humanized 11H1, humanized 6C5, and an isotype control. Color development was performed on a CytoFlex cytometer using a PE-labeled mouse anti-human Fc mAb, and values ​​corresponded to mean fluorescence intensity (MFI) (Figure 13). Results: As shown in Figures 11 to 13, the binding activity of different humanized anti-CLEC1 antibodies of the present invention to CLEC1-His, measured by ELISA, showed binding activity for all antibodies. All humanized anti-CLEC-1A antibodies of the present invention induce specific binding activity for CLEC-His. The binding activity of chimeric anti-CLEC1 antibodies to CLEC1-His, measured by ELISA, showed binding activity for all antibodies with different EC50s (Figure 12A). All humanized anti-CLEC-1A antibodies of the present invention induce specific binding activity for Fc-CLEC-1A with different ED50s (Figure 12B). Furthermore, all humanized antibodies were able to bind to human U266 cells (Figure 13). This example demonstrates that the antibodies of the invention have specific affinity for human CLEC-1A. To assess the binding ability of anti-CLEC1 antibodies with the same CDR combinations as the antibodies of the invention, referred to as 6C5, 11H11, and 14H9, mutated versions of these antibodies were provided and their binding ability to Fc-CLEC was assessed (FIGS. 14A-14D). The antibodies 6C5, 11H11, and 14H9 of the invention were mutated in the light chain variable region, or in the heavy chain variable region, or in the framework regions of the light and heavy chain variable regions. To that end, a mutated version of 14H9 (referred to as 14H9m in FIG. 14A) was prepared, comprising the light chain of 14H9 of SEQ ID NO: 7 and a mutated version of the heavy chain of SEQ ID NO: 6 (two substitutions in the third framework region compared to 14H9: T74S and V89I). A mutant version of 11H11 (referred to as 11H11m in Figure 14B) was prepared, which included the heavy chain of 11H11 (SEQ ID NO: 3) and a mutant version of the variable light chain of 11H11 (SEQ ID NO: 102) (a substitution in the second framework region compared to 11H11: A43S). Two mutant versions of 6C5 (referred to as 6C5m1 and 6C5m2 in Figures 14C-14D) were prepared. 6C5m1 includes the light chain of 6C5 (SEQ ID NO: 9) and a mutant version of the heavy chain (SEQ ID NO: 103) (a substitution in the third framework region compared to 6C5: M81I). 6C5m2 includes the heavy chain of 6C5 (SEQ ID NO: 8) and a mutant version of the light chain (SEQ ID NO: 104) (two substitutions in the second framework region compared to 6C5: A43P and R45K). The binding ability of these four mutants to Fc-CLEC1 was compared with that of their corresponding parent antibodies. As shown in Figure 14, the mutant antibodies have the same binding ability as their corresponding parent antibodies. Therefore, mutations in the framework regions may not affect the antibody's potency, provided that the CDRs are not modified.

Claims

1. Specifically binds to the extracellular domain of the human C-type lectin-like receptor-1 member A receptor (CLEC-1A receptor): an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 10, a VHCDR2 of SEQ ID NO: 11, a VHCDR3 of SEQ ID NO: 12; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 13, a VLCDR2 of SEQ ID NO: 14, a VLCDR3 of SEQ ID NO: 15; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 16, a VHCDR2 of SEQ ID NO: 17, a VHCDR3 of SEQ ID NO: 18; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 19, a VLCDR2 of SEQ ID NO: 20, a VLCDR3 of SEQ ID NO: 21; or an antibody heavy chain variable domain comprising a VHCDR1 of SEQ ID NO: 22, a VHCDR2 of SEQ ID NO: 23, and a VHCDR3 of SEQ ID NO: 24; and an antibody light chain variable domain comprising a VLCDR1 of SEQ ID NO: 25, a VLCDR2 of SEQ ID NO: 26, and a VLCDR3 of SEQ ID NO: 27 An antibody or antigen-binding fragment thereof comprising:

2. an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 3; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 4, or an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 5; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 7, or an antibody heavy chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 8; and an antibody light chain variable domain comprising or consisting of the amino acid sequence set forth in SEQ ID NO:

9. The antibody or antigen-binding fragment thereof of claim 1, comprising:

3. The antibody or antigen-binding fragment thereof of claim 1, which antagonizes the binding of the extracellular domain of human CLEC-1A to secondary necrotic cells and / or tumor cells and / or to the intracellular contents of secondary necrotic cells and / or tumor cells.

4. The antibody or antigen-binding fragment thereof according to claim 1, which antagonizes the binding of a fusion protein comprising the extracellular domain of the human CLEC-1A receptor fused to an Fc fragment of a human immunoglobulin to secondary necrotic cells and / or tumor cells and / or to the intracellular contents of secondary necrotic cells and / or tumor cells.

5. The antibody or antigen-binding fragment thereof according to claim 4, wherein the human immunoglobulin is human IgG.

6. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody is a recombinant antibody comprising a human IgG1, IgG2, IgG3, or IgG4 constant region.

7. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody heavy chain constant region comprises or consists of the amino acid sequence set forth in SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, SEQ ID NO: 97, SEQ ID NO: 100, or SEQ ID NO:

101.

8. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody light chain constant region is derived from or is a kappa light chain constant region.

9. 9. The antibody or antigen-binding fragment thereof according to claim 8, wherein the antibody light chain constant region comprises or consists of the amino acid sequence set forth in SEQ ID NO:

33.

10. The antibody or antigen-binding fragment thereof according to claim 1, which binds to human CLEC-1A with an affinity constant (KD) of at least 1E-07M.

11. The antibody or antigen-binding fragment thereof according to claim 10, which binds to human CLEC-1A with an affinity constant (KD) of at least 1E-08M.

12. The antibody or antigen-binding fragment thereof described in claim 1, which, when used in vivo and / or in vitro, correlates with modulation of phagocytosis of tumor cells and / or secondary necrotic cells by bone marrow cells compared to a negative control.

13. The antibody or antigen-binding fragment thereof of claim 12, wherein the modulation is an increase.

14. The antibody or antigen-binding fragment thereof according to claim 12, wherein the bone marrow cells are dendritic cells and / or macrophages.

15. The antibody or antigen-binding fragment thereof of claim 12, wherein phagocytosis of tumor cells is increased by at least 10% compared to a negative control.

16. A nucleic acid molecule or a combination of nucleic acid molecules encoding a polypeptide comprising or consisting of the antibody or antigen-binding fragment thereof of claim 1, wherein the nucleic acid molecule or combination of nucleic acid molecules encodes at least six CDR domains of the antibody or antigen-binding fragment thereof of claim 1.

17. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to claim 1 and / or a nucleic acid molecule or a combination of nucleic acid molecules according to claim 16, together with a pharmaceutically suitable vehicle.

18. A combination of compounds comprising a first therapeutic agent and at least one second therapeutic agent, i) the first therapeutic agent is an antibody or an antigen-binding fragment thereof according to claim 1, and / or a nucleic acid molecule or a combination of nucleic acid molecules according to claim 16; and ii) the at least one second therapeutic agent is selected from the list consisting of immunotherapeutic agents, chemotherapeutic agents, cell therapy agents such as CAR-T cells, and radiotherapy agents; A combination of compounds.

19. 19. The combination of claim 18, wherein the immunotherapeutic agent is a tumor-targeting antibody or an antigen-binding fragment thereof.

20. 20. The combination of claim 19, wherein the tumor-targeting antibody or antigen-binding fragment thereof is a tumor-targeting monoclonal antibody or antigen-binding fragment thereof.

21. The combination according to claim 20, wherein the tumor-targeting monoclonal antibody or antigen-binding fragment thereof is a tumor-targeting monoclonal antibody or antigen-binding fragment thereof that activates and / or enhances the phagocytic ability of macrophages.

22. 19. The combination of claim 18, wherein the immunotherapeutic agent is a monoclonal antibody selected from the group consisting of alemtuzumab, atezolizumab, bevacizumab, cetuximab, herceptin, panitumumab, rituximab, trastuzumab, anti-PDL-1 antibody, and anti-CD47 antibody, and / or a monoclonal antibody selected from the group consisting of anti-PD1 antibody, anti-CTLA4 antibody, agonist anti-CD137 antibody, anti-CD28 antibody, anti-CD127 antibody, anti-bcl2 antibody, and anti-SIRPa antibody.

23. 19. The combination of claim 18, wherein the radiotherapeutic agent is a cytotoxic agent with antiproliferative, proapoptotic, cell cycle arresting, and / or differentiation-inducing effects.

24. 24. The combination of claim 23, wherein the cytotoxic agent having an antiproliferative, pro-apoptotic, cell cycle arresting, and / or differentiation-inducing effect is a cytotoxic agent selected from the group consisting of cytotoxic antibodies, alkylating agents, anthracyclines, antimetabolites, anti-microtubule agents, topoisomerase inhibitors, alkaloids, bleomycin, anti-neoplastic agents, and cyclophosphamide.

25. 19. The combination of claim 18 for simultaneous, separate or sequential use of the first and second therapeutic agents.

26. 17. An antibody or antigen-binding fragment thereof according to claim 1, or a nucleic acid molecule or a combination of nucleic acid molecules according to claim 16, for use as a medicament.

27. An antibody or antigen-binding fragment thereof described in claim 1, or a nucleic acid molecule or a combination of nucleic acid molecules described in claim 16, for use in the prevention and / or treatment of a disease or disorder in which increased phagocytic ability by bone marrow cells ameliorates or prevents the disease or disorder.

28. 28. The antibody or antigen-binding fragment thereof, or the nucleic acid molecule or combination of nucleic acid molecules of claim 27, wherein the disease or disorder is a human disease or disorder.

29. The antibody or antigen-binding fragment thereof, or the nucleic acid molecule or combination of nucleic acid molecules according to claim 27, wherein the bone marrow cells are dendritic cells and / or macrophages.

30. An antibody or antigen-binding fragment thereof described in claim 1, or a nucleic acid molecule or a combination of nucleic acid molecules described in claim 16, for use in the treatment of a disease or condition, wherein induction of phagocytosis in a patient ameliorates or prevents the disease or condition.

31. An antibody or antigen-binding fragment thereof described in claim 1, or a nucleic acid molecule or a combination of nucleic acid molecules described in claim 16, for the treatment of patients with cancer, inflammatory disease, chronic infection, or sepsis.

32. The antibody or antigen-binding fragment thereof, or the nucleic acid molecule or combination of nucleic acid molecules described in claim 31, wherein the cancer is a liquid cancer or a solid cancer.

33. The antibody or antigen-binding fragment thereof, or the nucleic acid molecule or combination of nucleic acid molecules according to claim 31, wherein the cancer is lymphoma, colorectal cancer, mesothelioma, or liver cancer.

34. 17. The antibody or antigen-binding fragment thereof of claim 1, or the nucleic acid molecule or combination of nucleic acid molecules of claim 16, for use in combination therapy in which a first pharmaceutical agent comprising a chemotherapeutic agent, a radiotherapeutic agent, an immunotherapeutic agent such as a tumor-targeting monoclonal antibody, a cell therapy agent such as CAR-T cells, an immunosuppressant, an apoptotic promoter, an antibiotic, a targeted cancer therapy, and / or a probiotic is administered to a patient in need thereof.

35. 35. The antibody or antigen-binding fragment thereof, or nucleic acid molecule or combination of nucleic acid molecules according to claim 34, for use in combination therapy in which a first medicament comprising a chemotherapeutic agent, a radiotherapeutic agent, an immunotherapeutic agent such as a tumor-targeting monoclonal antibody, a cell therapy agent such as CAR-T cells, an immunosuppressant, an apoptotic promoter, an antibiotic, a targeted cancer therapy, and / or a probiotic is administered simultaneously, separately, or sequentially to a patient in need thereof.

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