Anti-chemokine-like receptor 1 antibodies and therapeutic uses thereof

Abstract

The present invention relates to anti-chemokine-like receptor 1 antibodies and therapeutic uses thereof. The present invention provides anti-CMKLR1 compounds having agonist ability for the interaction between Resolvin E1 and CMKLR1, and their use in the treatment or prevention of diseases, particularly diseases in which inflammation regression is delayed or disrupted.

Description

[0001] This application is a divisional application. The application date of the original application is April 3, 2019, the application number is 201980037001.4 (PCT / EP2019 / 058358), and the name of the invention is “Anti-chemokine-like receptor 1 antibodies and their therapeutic applications”. Technical Field

[0002] The present invention relates to the field of immunotherapy. It provides novel anti-chemokine receptor antibodies that have agonist activity against chemokine-like receptor 1 (CMKLR1). It also provides the use of such antibodies in therapy, particularly for the treatment of autoimmune and chronic inflammatory diseases, infectious diseases, cancer, and any condition in which the resolution phase of inflammation is disrupted or delayed. Background Art

[0003] The key role of inflammatory processes in health and disease has long been known. The detailed molecular mechanisms and biological events that regulate the progression and resolution of inflammation remain crucial. Recent studies have provided strong evidence that the resolution of inflammation is not a passive process as previously believed. Instead, the resolution of inflammation is a biosynthetically active process that is regulated by biochemical mediators and receptor signaling pathways. Therefore, resolution is driven by specific pro-resolution mediators. Inflammation is an autonomous mechanism that occurs during infection, injury or trauma. Inflammation is inevitable and usually beneficial, and the inflammatory response is coordinated by a delicate balance between positive and negative feedback loops. Inflammation is generally divided into three steps: initiation, amplification and resolution.

[0004] The initiation step is characterized by vasodilation. Resident cells (dendritic cells (DCs) and macrophages) recognize infecting pathogens or danger signals. This step induces the secretion of cytokines and chemokines, as well as the production of proinflammatory lipid mediators called prostaglandins (PGs) and leukotrienes. Chemokines return to the bloodstream and trigger the recruitment of innate immune cell actors. The amplification step begins with the recruitment of inflammatory cells of the immune system. The first actors recruited to the site of infection are polymorphonuclear granulocytes (PMNs). During the cellular phase, PMNs recognize pathogens and take on the task of eliminating them. Other cells, such as DCs and proinflammatory M1 macrophages, arrive at the site of inflammation to assist PMNs. DCs enter the bloodstream and travel to lymph nodes to activate adaptive immunity. T and B lymphocytes arrive at the site of inflammation and eliminate infected cells. Once the pathogen is completely eliminated, it becomes crucial to halt inflammation to prevent chronic inflammation, which can lead to disease. To achieve this, an active mechanism known as resolution occurs. Therefore, resolution is described as the final step of inflammation.

[0005] The resolution process that leads to the termination of an inflammatory response is a complex one involving the sequential and temporal involvement of cellular (eg, granulocytes or macrophages) and chemical (eg, cytokines or specific prolytic mediators or factors) effectors.

[0006] Defective resolution may result in increased infiltration of granulocytes into the inflammatory site (e.g., as measured by histology, cell counting, or indirect biochemical techniques, such as quantification of elastase by enzyme-linked immunosorbent assay for granulocyte receptor 1 or molecular quantification of granulocyte receptor 1 by PCR) and delayed apoptosis of such cells (e.g., as measured cytologically using antibodies specific for annexin 5). Defective resolution may also result in sustained or increased synthesis of proinflammatory cytokines (e.g., TNF-α, IL-8, or IL-12) and decreased production of anti-inflammatory cytokines (e.g., IL-10) (as measured by enzyme-linked immunosorbent assay or PCR) and sustained or increased activation of transcription factors involved in inflammatory cytokine synthesis (e.g., NF-κB) (e.g., as measured by nuclear translocation or Western blot and quantification of IκB degradation levels). Specific pro-resolution mediators (e.g., lipoxins, resolvins, protectins, or maresins) or their precursors (e.g., 17-HDOHE or 14-HDOHE) can also be quantified by mass spectrometry or enzyme-linked immunosorbent assays. Resolution defects, in turn, result in defects in the synthesis of one or more of these mediators. Resolution defects may also be due to reduced expression of receptors for resolvable molecules (ALX / CMK1R1, GPR32, GPR18), or internalization of these receptors into the cytoplasm and processing, or overexpression of receptors for certain inflammatory cytokines or lipids. These conditions can be measured by histology, cytology, or PCR. Resolution defects may also result in reduced or inhibited conversion of M1 macrophages to M2 macrophages, and impaired phagocytosis or efferocytosis of the same cells.

[0007] It is now clear that in addition to the checkpoint activators and / or inhibitors involved in this mechanism, a family of chemicals also actively promotes the resolution of inflammation and tissue repair without compromising host defenses. Events during acute inflammatory episodes establish biosynthetic pathways for a range of chemical mediators that act as antagonists and agonists, meaning that they not only inhibit inflammatory pathways but also strike inflammation, thereby restoring tissue homeostasis and function. Therefore, anti-inflammatory factors are not equivalent to pro-resolving factors (Buckley et al., 2014; Serhan, 2014a). Failure to resolve acute inflammation is associated with the development of chronic inflammation. Therefore, anti-inflammatory compounds refer to inhibitors or blockers of inflammatory resolution, molecules that prevent immune extravasation; whereas pro-resolving factors stimulate and / or activate specific processes that trigger or enhance inflammatory resolution (such as apoptosis or efferocytosis).

[0008] Resolution begins shortly after the initiation of an inflammatory response initiated by PMNs. They eliminate pathogens and simultaneously begin synthesizing specific pro-resolving mediators (SPMs). These SPMs are key players in the resolution phase. Neutrophils control the onset of the resolution phase by activating pro-resolving circuits to ensure the safe conclusion of the inflammatory response. During the early stages of resolution, neutrophils undergo a phenotypic switch, producing different lipid mediators depending on the cells and substrates present in the environment. PMN-LTs switch to the PMN-LO (lipoxygenase) pathway under the influence of lipoxins and resolvins. They are exposed to a gradient of autologous autacoids, which initiates this phenotypic change. Lipoxins are produced through biosynthetic pathways involved during cell-cell interactions (PMN-5-LO / tissue-resident cells – 5-LO). Wound healing has been observed to be delayed in models of neutrophil depletion, with neutrophils releasing proteases that inactivate inflammatory cytokines.

[0009] Various molecules participate in the initiation or inhibition of inflammation regression.The following molecules are examples of such active molecules.COX-2 (cyclooxygenase 2), a kind of prostaglandin-endoperoxide synthase (PTGS), is the enzyme responsible for forming prostaglandins (including thromboxane and prostaglandin, such as prostaglandin), has the dual action that causes inflammatory attack, and afterwards as the helper of regression process.COX-2 inhibitors may play a beneficial role in the early stage of inflammation.COX-2 inhibitors also have harmful consequences, such as reducing early PMN transport, destroying the generation of LXA, reducing the phagocytosis of macrophages and reducing PGE2 and LXA.In the inflammation regression process, negative feedback also relates to the anti-inflammatory prostaglandin called PGD2 / 15dPGJ, but other prostaglandins (such as PGE2) also participate in positive feedback.

[0010] G protein-coupled receptors (GPCRs) constitute a large family of proteins that encompass a wide range of functions, including various autocrine, paracrine, and endocrine processes. They display considerable diversity at the sequence level, allowing them to be divided into distinct groups. Typically, activation of GPCRs engages a broad network of signaling pathways mediated by G proteins or B-arrestins, or both.

[0011] Chemokine-like receptor 1 (CMKLR1) (also known as ChemR23) and chemokine-like receptor 2 (CCRL2) are seven-transmembrane receptors that are recognized by known G protein-coupled receptors with which they are homologous (AJ Kennedy and AP Davenport, 2018). Chemokine-like receptor 1 (CMKLR1; also known as Dez in murine species) is an orphan G protein-coupled receptor related to GPR-1 (38% overall amino acid identity), C3a receptor (38%), C5a anaphylatoxin receptor (36%), and formyl Met-Leu-Phe receptor (35%). ChemR23 is more distantly related to the chemokine receptor subfamily (Samson et al., 1998). CMKLR1 is expressed on monocytes, macrophages, dendritic cells, and NK cells, as well as adipocytes and endothelial cells. CMKLR1 expression has been described in many cell populations beyond leukocytes, including preadipocytes and adipocytes (Goralski et al., 2007; Roh et al., 2007), skeletal muscle cells (Sell et al., 2009), and endothelial cells (Kaur et al., 2010), with additional roles for the chemerin / CMKLR1 system proposed in controlling lipid and glucose metabolism (Bozaoglu et al., 2007; Ernst and Sinal, 2010), blood pressure (Watts et al., 2013), and angiogenesis (Kaur et al., 2010).

[0012] Recent studies have begun to uncover the ligands for these receptors and their functions. Thus, the plasma protein-derived chemoattractant chemokines are ligands for CMKLR1, and activation of CMKLR1 with chemokines has been shown to induce migration of macrophages and dendritic cells (DCs) in vitro, suggesting a pro-inflammatory role for chemokines. Conversely, in vivo studies using CMKLR-deficient mice have shown that these receptors may have anti-inflammatory effects, possibly due to the recruitment of plasmacytoid DCs. Chemokine / CMKLR1 interactions also promote adipogenesis and angiogenesis.

[0013] Chemokines act as chemokines during inflammation, recruiting cells to sites of infection. These ligands promote the resolution of inflammation in animal models of acute inflammation by enhancing PMN apoptosis and M2-dependent efferocytosis (a non-inflammatory phagocytosis without the release of proinflammatory mediators), as well as by reducing DC migration and secretion of IL-12 (a proinflammatory cytokine) (Serhan, 2014b).

[0014] Chemokines are abundant in inflammatory fluids and possess antimicrobial activity. They have been shown to attract leukocytes expressing CMKLR1 and promote macrophage adhesion to extracellular matrix proteins (Wittamer et al., 2003). Furthermore, chemokines have recently been identified as adipokines. Chemokines are secreted by mature adipocytes and stimulate preadipocyte differentiation. Elevated serum levels of chemokines are associated with chronic inflammatory diseases, coronary artery disease, metabolic syndrome, and obesity. There are indications that the increased production of chemokines in obese adipose tissue may lead to increased macrophage infiltration in obese adipose tissue, thereby reducing inflammation. When chemokines bind to CMKLR1, two signaling pathways are activated: the G protein signaling pathway and the β-arrestin signaling pathway.

[0015] The second ligand for CMKLR1 is the lipid mediator resolvin E1 (RvE1), which belongs to the resolvin family. The anti-inflammatory lipid mediator resolvin E1 inhibits leukocyte infiltration and proinflammatory gene expression. These diverse results suggest that CMKLR1 is a multifunctional receptor. It is involved in increasing PMN apoptosis and M2 efferocytosis and reducing DC migration and secretion of proinflammatory cytokines such as IL-12. RvE1 is produced by neutrophils and endothelial cells. Aspirin treatment induces RvE1 in vitro, as aspirin activates the COX-2 pathway responsible for RvE1 production. Furthermore, neutrophils are able to convert 18R-HEPE to RvE1. Reduced phagocytosis by macrophages has been observed in patients with localized aggressive periodontitis (LAP). RvE1 rescues the impaired phagocytic activity of LAP macrophages. RvE1 also enhances resolution by reducing IL-23 and IL-6 and increasing IFN-γ in the allergic airways of mice. RvE1 regulates natural killer (NK) cell migration and cytotoxicity. When RvE1 binds to CMKLR1, only the G protein signaling pathway is activated, while the B-arrestin pathway is inhibited. Under specific conditions, the B-arresting pathway is inhibited.

[0016] Initially, attention to the chemokine system focused on its role in inflammation and chemotaxis of immune cells following its discovery in psoriasis. More recently, given the role of the chemokine system in inflammation, obesity, and metabolic syndrome, its potential role in cardiovascular function and in reproductive biology has been studied. Therefore, the chemokine system has attracted widespread attention due to its role in inflammatory processes (particularly its role in inflammatory resolution). Many diseases are associated with a delay or interruption in the resolution process. Most of the currently known specific pro-resolving factor mediators are derived from polyunsaturated fatty acids (including lipoxins), and the resolving family includes E-series and D-series resolvins, protectins, and maresins. However, pro-resolving molecules are difficult to synthesize due to their lipid properties. For example, it is difficult to produce sufficient amounts of pro-resolving molecules, such as for clinical trials, and SPMs are rarely produced effectively. In addition, it is difficult to produce antibodies that specifically target G protein-coupled receptors. Therefore, there is a need for molecules (such as pro-resolving factors) that have the ability to trigger or enhance the resolution phase of the inflammatory response. Summary of the Invention

[0017] In a first aspect, the present invention relates to an anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic or a modified antibody. In the following disclosure, the anti-CMKLR1 compound is considered to be an anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic or a modified antibody. In a specific embodiment of the invention, the compound is defined by the sequence of its CDRs. In a more specific embodiment of the invention, the anti-CMKLR1 compound is an antibody defined by the sequence of its CDRs and framework regions (FRs). The anti-CMKLR1 compound is a compound that specifically binds to chemokine-like receptor 1 (CMKLR1). In the following disclosure, the terms "chemokine-like receptor 1", "CMKLR1" and "ChemR23" are used interchangeably and all refer to the receptor encoded by the gene CMKLR1 in humans or the gene cmklr1 in non-human animals. In a specific embodiment of the invention, the anti-CMKLR1 compound specifically binds to human CMKLR1, or in other words, the present invention relates to an anti-human CMKLR1 compound. As used herein, the term "CMKLR1" refers to the chemokine-like receptor 1 protein (also known as chemR23), which is a member of the G protein-coupled receptor family from mammalian species, preferably human CMKLR1. The reference sequence of the human CMKLR1 protein used in the examples of this application corresponds to the sequence associated with Uniprot accession number Q99788 (SEQ ID NO: 1).

[0018] In a second aspect, the present invention relates to an anti-CMKLR1 antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody defined by at least one functional characteristic. In a preferred embodiment, the anti-CMKLR1 compound is defined by its ability to inhibit the secretion of pro-inflammatory cytokines (particularly IL-12) and / or enhance the secretion of anti-inflammatory cytokines (particularly IL-10 and / or CCL17). In a more specific embodiment, the anti-CMKLR1 compound inhibits or enhances cytokine secretion by macrophages (particularly M1 macrophages and / or M2 macrophages). In a specific embodiment, the anti-CMKLR1 compound of the present invention enhances the polarization of macrophages to anti-inflammatory macrophages (particularly M2 macrophages).

[0019] In a third aspect, the present invention relates to an anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic having agonist properties against resolvin E1 (RvE1), thereby simulating the binding of RvE1 to CMKLR1 on CMKLR1-positive cells. "Agonist properties against RvE1-CMKLR1 interaction" refers to an antibody or antigen-binding fragment thereof or an antigen-binding antibody mimetic or modified antibody of the present invention that targets CMKLR1 and has the effect of simulating the binding of RvE1 to CMKLR1 (especially on dendritic cells, monocytes and macrophages) (especially the binding of human RvE1 to human CMKLR1), thereby activating the receptor signaling pathway normally activated by RvE1. Since the binding and activation of the receptor will produce a biological response, the compounds of the present invention can lead to the G protein signaling pathway (especially Gα i and / or Gα o signaling pathway) without activating the β-arrestin pathway; in particular, the compounds of the present invention can lead to inhibition of the β-arrestin pathway. In particular, the binding of the compounds of the present invention induces activation of Akt and / or Erk proteins in vitro and / or in vivo. In other words, compared to a control antibody, a resolvin E1-like agonist antibody can be defined as an antibody that can bind to CMKLR1 and thereby induce phosphorylation of Akt and / or Erk proteins. The control antibody can be an antibody that does not specifically bind to CMKLR1. Protein phosphorylation can be determined according to methods known to those skilled in the art (e.g., the methods disclosed in the Examples of this specification).

[0020] In one embodiment, the compound of the present invention is an RvE1-like agonist, that is, the compound of the present invention is an agonist of the CMKLR1 signaling pathway induced by RvE1. In other words, the anti-CMKLR1 compound of the present invention is an agonist of the interaction between RvE1 and CMKLR1 (particularly human RvE1 and human CMKLR1). In one embodiment, the compound of the present invention enhances the activation of the G protein pathway induced by CMKLR1. In another embodiment, the compound of the present invention does not induce the activation of the β-arrestin pathway induced by CMKLR1. In another embodiment, the compound of the present invention inhibits the β-arrestin pathway induced by CMKLR1. In another embodiment, since the compound of the present invention induces at least one agonist effect of RvE1 binding to CMKLR1 and RvE1 is a pro-resolving factor or pro-resolving mediator, the compound of the present invention is a pro-resolving factor or pro-resolving mediator.

[0021] In one embodiment, the compounds of the present invention do not interfere with the binding of chemokines to CMKLR1. Chemokine is one of the natural ligands of CMKLR1. In other words, the compounds of the present invention are not agonists of the interaction between chemokines and CMKLR1. The absence of such agonist activity can be assessed according to embodiments of the present invention, wherein a competition assay is disclosed that measures chemokine-dependent B-arrestin recruitment through the CMKLR1 receptor in the presence of an anti-CMKLR1 antibody of the present invention. In a preferred embodiment, the anti-CMKLR1 compounds of the present invention do not compete with chemokines for binding to CMKLR1. The absence of competition between the anti-CMKLR1 compounds of the present invention and chemokines can be determined by determining that the binding of chemokines to CMKLR1 in the presence of a CMKLR1 compound of the present invention is at least 50%, preferably at least 80%, more preferably at least 90%, and most preferably similar to the binding of chemokines to CMKLR1 under the same experimental conditions but in the absence of an anti-CMKLR1 antibody of the present invention. Alternatively, the absence of competition between the anti-CMKLR1 compounds of the present invention and chemokines can be determined according to the method described in Example 11.

[0022] In one embodiment, the anti-CMKLR1 compound has the ability to activate at least one Akt signaling pathway protein (also known as the PI3K-Akt pathway) and / or Erk signaling pathway protein (preferably Akt protein and / or Erk protein, preferably both Akt and Erk proteins) in vitro and / or in vivo. Pathway activation can be assessed according to methods known in the art, particularly using the methods disclosed in the Examples of the present invention.

[0023] In one embodiment, the antibodies, antigen-binding fragments thereof, antigen-binding antibody mimetics, or modified antibodies of the present invention enhance RvE1-induced IL-10 cytokine secretion in vitro and / or in vivo (particularly macrophage IL-10 cytokine secretion). In one embodiment, the antibodies, antigen-binding fragments thereof, antigen-binding antibody mimetics, or modified antibodies of the present invention enhance RvE1-induced IL-10 and CCL17 cytokine secretion in vitro and / or in vivo (particularly macrophage IL-10 and CCL17 secretion). In one embodiment, the antibodies, antigen-binding fragments thereof, antigen-binding antibody mimetics, or modified antibodies of the present invention inhibit IL-12 cytokine secretion in vitro and / or in vivo (particularly macrophage IL-12 secretion). The inhibitory effect may be partial inhibition, i.e., in the presence of an anti-CMKLR1 compound, the secretion level of IL-12 is reduced to below baseline levels (i.e., the level in the absence of RvE1 or anti-CMKLR1 compound), or the inhibitory effect may be complete inhibition (no IL-12 secretion). BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : Effects of anti-CMKLR1 variants 1G1, 2G1, 3G1 and 4G1 chimeric antibodies on DC maturation and differentiation. Mouse dendritic cells were incubated in the maturation stage (24 hours or 48 hours), and then the differentiation stage was performed using vehicle or RvE1 or anti-CMKLR1 antibody variants (mutations at positions 1 and 2 of SEQ ID NO.8, corresponding to VH-CDR3: 1G1 (LL), 2G1 (LI), 3G1 (IL) or 4G1 (II)) or isotype control (hIgG1 or mlgG). The cells were then stained with cell marker antibodies for FACS analysis: A. CD80-PE; B. CD86-FITC; C. CD103-PerCPCy5.5; D. 1 / Ab-APC. The mean fluorescence under each condition was determined. E and F represent the fluorescence of each condition using Life Technologies after 24 hours or 48 hours of maturation, respectively. Kit for measuring cell viability by FACS.

[0025] Figure 2 Effects of anti-CMKLR1 antibodies on a DSS-induced acute inflammatory colitis model in mice. Six days after DSS induction, mice were injected three times daily for five days with isotype control hIgG1 (10 μg / mouse) (x), RvE1 (10 μg / mouse) (●), or 2G1 antibody (10 μg / mouse) (□). Untreated wild-type mice are indicated by ▼. A. Animal weight loss. B. Animal fecal score. C. Colon length. D. Collapse index.

[0026] Figure 3 Effect of anti-CMKLR1 antibodies on a TNBS-induced acute inflammatory colitis model in mice. Mice received 200 μL of 5% haptenating agent TNBS (in 50% ethanol) on day 0, followed by three daily injections of isotype control hIgG1 (10 μg / mouse) (x), RvE1 (1 μg / mouse) (●), or 2G1 antibody (10 μg / mouse) (□) for 5 days, or were left untreated (wild-type animals) (▲). Mice were sacrificed, and colon length was measured under each condition.

[0027] Figure 4 Effects of anti-CMKLR1 antibodies on a chronic inflammatory colitis model in IL-10KO mice. IL-10KO mice developed spontaneous inflammatory colitis. These mice were treated intraperitoneally with an anti-CMKLR1 antibody (2G1) (□) or an isotype control (hIgG1) (x) (25 μg / injection, three times weekly). A. Animal weight loss during and after treatment. B. Animal fecal scores.

[0028] Figure 5 Effects of anti-CMKLR1 antibodies on a mouse model of type 1 non-obese diabetes. Mice developed spontaneous diabetes. When blood glucose levels ranged from 180 mg / dL to 234 mg / dL, anti-CMKLR1 antibodies (□ in A and B, ■ in C) or an isotype control antibody (x in A and B; □ in C) were administered intraperitoneally at 20 μg / injection three times weekly for two weeks. A. Percent survival; B. Individual blood glucose concentrations (mg / dL). C. Total blood glucose concentrations (mg / dL).

[0029] Figure 6 Effects of Resolvin E1 on autoimmune diseases (e.g., a mouse psoriasis model). Aldara-treated mice were intraperitoneally injected daily for 6 consecutive days with RvE1 (1 μg) (●) and PBS, and an isotype control antibody (10 μg) (x) was injected on days 0, 2, and 4. A. Ear thickness and B. Skin thickness were measured daily until recovery.

[0030] Figure 7 Effects of anti-CMKLR1 antibodies on a mouse peritonitis model. Preventive injections of anti-CMKLR1 (1 mg (1 mL) / mouse), RvE1 (1 μg / mouse) (●), 2G1 antibody (10 μg / mouse) (□), or an isotype control antibody (x) were administered prior to Zymosan A injection. A. PMN counts within the first 50 hours after Zymosan A injection. B. Macrophage counts within the first 50 hours after Zymosan A injection. C. Resolution index.

[0031] Figure 8 Effects of anti-CMKLR1 antibody on the 4T1 mammary tumor model. Mice were inoculated with 250,000 4T1 cells in the mammary gland. Anti-CMKLR1 antibody (2G1) was then injected twice (10 μg / injection) on days 4 and 7. Or anti-41BB antibody (3H3) (●) or both antibodies (▲), or control antibody (IgG1 isotype antibody clone 3G8) (x A. and B.) were injected three times a week for 3 weeks. A. Tumor area was measured 8 days after tumor injection and then every 2 days. B. Tumor lung metastasis was measured by bioluminescence imaging (BLI) in animals (n=4) treated with isotype control and anti-CMKLR1 antibodies.

[0032] Figure 9 Effects of anti-CMKLR1 antibodies on two different mouse models of colon cancer. Both mouse models were studied. Animals were injected intraperitoneally with either an isotype control antibody (3G8) (x) or anti-CMKLR1 (2G1) (□) at 20 μg / injection for three weeks. A-C. Results in the CT26 colon cancer model compared single treatment with p84 (A) and 2G1 (B) to combination treatment with these two antibodies (C). D. Results in the MC38 colon cancer model.

[0033] Figure 10 : CMKLR1 expression in biopsies of patients with ulcerative colitis (UC) or Crohn's disease before and after anti-TNFα treatment. x indicates control; =A represents CMKLR1 expression in patients who responded to corticosteroids and / or immunosuppressive therapy before infliximab treatment; ■ represents CMKLR1 expression in patients who did not respond to corticosteroids and / or immunosuppressive therapy before infliximab treatment; Δ represents CMKLR1 expression in patients who responded to corticosteroids and / or immunosuppressive therapy after infliximab treatment; ▲ represents CMKLR1 expression in patients who did not respond to corticosteroids and / or immunosuppressive therapy after infliximab treatment. A. CMKLR1 transcript expression in patients with ulcerative colitis before and after infliximab treatment. B. CMKLR1 transcript expression in inflamed colon biopsies from patients with Crohn's disease before and after infliximab treatment.

[0034] Figure 11Figure 2: CMKLR1 expression in UC patient biopsies before and after anti-α4β7 (VDZ) therapy. A. Represents CMKLR1 transcript expression in inflamed colon biopsies from ulcerative colitis patients before and after vedolizumab treatment. R corresponds to patients who responded to VDZ treatment, and NR corresponds to patients who did not respond to VDZ treatment.

[0035] Figure 12 : Binding of anti-CMKLR1 antibodies to CMKLR1 peptides was determined by ELISA. Binding of 2G1 antibody (□) on the CMKLR1 EL3 loop (SEQ ID NO: 18) was compared to an isotype control antibody (3G8) (x) by ELISA measuring OD at 450 nm.

[0036] Figure 13 CMKLR1 expression in various cell lines was investigated by FACS and Western blot. A. CMKLR1 cell surface protein expression was measured using two different human T cell lines, Thp1 and U937, using the 2G1 antibody at various concentrations (ng / ml). B. CMKLR1 protein expression was measured by Western blot in T cell lines (Thpl and U937), the fibroblast line MRC5, the NK cell line NKL, and CMKLR1-negative and transduced CHO cells.

[0037] Figure 14 CMKLR1 expression in mouse myeloid cells was assessed by FACS. Following differentiation, cell surface expression of CMKLR1 in mouse myeloid lineage cells was analyzed. A. Expression in macrophages and monocytes (MOs). B. and C. Expression in macrophages (mM1 and mM2). D. and E. Expression in dendritic cells (mDCs and iDCs).

[0038] Figure 15: Study of inflammatory cytokine secretion by human macrophages during CMKLR1 activation. After differentiation of human macrophages into M1 or M2 cells, cells were incubated in the presence of culture medium, isotype control, anti-CMKLR1 antibodies (H6 and BZ194), C15 peptide, 2G1 anti-CMKLR1 antibody, or RvE1. IL-10, CCL17, and IL-12p40 secretion were assessed by ELISA. Cytokine secretion in the supernatant was measured using BD ELISA kits. The supernatant was diluted 1 / 10 for detection of the IL-10 cytokine, 1 / 50 for detection of the CCL17 cytokine, and 1 / 100 for detection of the IL-12p40 cytokine. A. IL-10 cytokine secreted by M2 cells. B. CCL17 cytokine secreted by M2 cells. C. Represents IL-12 cytokine secreted by M1 cells.

[0039] Figure 16 : Study of inflammatory cytokine secretion by human macrophages during CMKLR1 activation. After differentiation of human monocytes, cells were incubated in the presence of different isotype controls (mlgG4; hIgG4 or hIgG1, 2 μg / ml), C7 commercial anti-CMKLR1 antibody (2 μg / ml), 2G4 or 2G1 antibody (2 μg / ml), or other commercial anti-CMKLR1 antibodies (H6 (2 μg / ml), BZ332 (2 μg / ml), or 84939 (2 μg / ml), C15 peptide (10 nM), or RvE1 (10 ng / ml). A. Expression of the CD200R marker was measured by FACS. B. IL-10 cytokine secreted by M2 cells. C. CCL17 cytokine secreted by M2 cells. D. Represents IL-12 cytokine secreted by M1 cells. Secretion of IL-10, CCL17, and IL-12p40 was assessed by ELISA.

[0040] Figure 17 Study of myeloid cell activation markers following CMKLR1 pathway activation. Dendritic cells were incubated with vehicle, RvE1, 2G1, or an isotype control (hIgG1) and then stained for FACS analysis using the following marker antibodies: A. CD80-PE; B. CD86-FITC; C. CD103-PerCPCy5.5; D. CD40-PeCy 7; E. 1 / Ab-APC. Mean fluorescence was measured for each condition.

[0041] Figure 18CMKLR1 pathway activation studies: Akt and Erk phosphorylation. Western blot analysis of ERK and AKT pathway activation was performed after incubation of mouse M1 cells with 2G1 or RvE1 for 5, 10, and 30 minutes. Phosphorylation of Akt or Erk was assessed using P-Akt or P-Erk (p44 / 42) antibodies. A. Erk and Akt activation by RvE1. B. Erk and Akt activation by 2G1.

[0042] Figure 19 Figure 3: CMKLR1 expression in human monocytes and mouse bone marrow cells stimulated with proinflammatory stimuli. CMKLR1 (ChemR23) expression was measured by FACS after 16 or 48 hours of stimulation with LPS, TNFα, or IL-6. A. Results for human blood monocytes; B. and C. Results for myeloid cells and neutrophils from mouse bone marrow.

[0043] Figure 20 : Competition study of the interaction between chemokine-CMLKRI and anti-CMKLR1 antibodies.

[0044] A. The inhibitory effects of chemokines from two different suppliers, DiscoverX (·) or R&D System (▲), or in combination with anti-CMKLR1 antibody (2G1) alone (◇) or in combination with chemokines from DiscoverX (□) on cAMP were examined.

[0045] B. Activation of beta arrestin in the presence of anti-CMKLR1 antibodies at different concentrations ranging from 1 μM to 1 nM and chemokine at 2 nM (◯) or 6 nM (·).

[0046] Figure 21 :Anti-CMKLR1 antibody in CD45Rb 高 Role of T cell transfer in a mouse model of chronic colitis.

[0047] Treated animals were followed for changes in body weight for up to 60 days.Animals were treated with isotype control hIgG1 (x) or anti-CMKLR1 antibody (■).

[0048] Figure 22:The effect of anti-CMKLR1 antibody on hepatocarcinoma mouse model (HCC model). Anti-CMKLR1 antibody (2G1, 0.8 mg / kg) was injected intraperitoneally (ip) 3 times a week (for a total of 2 weeks), with or without combination (·) of anti-PD1 mAb (RMP1-14 clone, 8 mg / kg) injected twice on day 4 and day 8, or anti-PD-1 antibody (Δ) injected alone (twice a week) to investigate the anti-tumor effect. In an orthogonal model of mouse HCC (on day 0, 2.5×10 6 Mice were treated in Hepa 1.6 cells for 2 weeks. An isotype control antibody was administered at a dose of 0.8 mg / kg three times weekly for 2 weeks. After stopping treatment, mice were considered to have a partial response (PR) if they survived for several days to 1 month; a complete response (CR) if they survived for more than 1 month; and a cure was considered if they survived three times longer than the time required for all control mice to be stained. DETAILED DESCRIPTION

[0049] As used herein, the term "antibody" includes polyclonal antibodies, monoclonal antibodies or recombinant antibodies. As used herein, compared to "polyclonal" antibody preparations comprising a mixture of antibodies with different amino acid sequences, "monoclonal antibody" means an antibody molecule preparation that obtains antibodies with a common heavy chain and a common light chain amino acid sequence. Monoclonal antibodies can be produced by several known techniques, such as phage, bacteria, yeast or ribosome display, and the classical method of hybridoma-derived antibodies as an example. Antibodies can also be synthesized using a disclosed amino acid sequence as a reference. Therefore, the term "monoclonal" is used to refer to all antibodies derived from a nucleic acid clone.

[0050] The antibodies of the present invention include recombinant antibodies. As used herein, the term "recombinant antibody" refers to an antibody generated, expressed, produced, or isolated by recombinant means, for example, an antibody expressed using a recombinant expression vector transfected into a host cell; an antibody isolated from a recombinant combinatorial antibody library; an antibody isolated from an animal (e.g., a mouse) transgenic for human immunoglobulin genes; or an antibody generated, expressed, produced, or isolated by any other means in which a specific immunoglobulin gene sequence (e.g., a human immunoglobulin gene sequence) is assembled with other DNA sequences. For example, recombinant antibodies include chimeric antibodies and humanized antibodies.

[0051] As used herein, "chimeric antibodies" refer to antibodies in which variable domain sequences derived from the germline of a mammalian species (eg, mouse) have been spliced onto constant domain sequences derived from the germline of another mammalian species (eg, human).

[0052] As used herein, in the first embodiment, a "humanized antibody" refers to such an antibody in which CDR sequences derived from the germline of another mammalian species (e.g., a mouse) are spliced onto human or humanized framework sequences. In another embodiment, a "humanized antibody" refers to an antibody in which at least one CDR and all or part of the framework sequence are humanized.

[0053] As used herein, an "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) that exhibits antigen-binding ability to CMKLR1, which may be in its native form; such a fragment, in particular, exhibits the same or substantially the same antigen-binding specificity for the antigen compared to the antigen-binding specificity of the corresponding 4-chain antibody. Advantageously, the antigen-binding fragment has a binding affinity similar to that of the corresponding 4-chain antibody. However, antigen-binding fragments with reduced antigen-binding affinity compared to the corresponding 4-chain antibody are also encompassed by 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 the antibody.

[0054] Antigen-binding fragments of antibodies are fragments that contain their hypervariable domains, known as CDRs (complementarity determining regions), or portions thereof that contain the antigen recognition site (i.e., the extracellular domain of CMKLR1, particularly the third loop of the extracellular domain of CMKLR1 (referred to as EL3)), thereby defining the specificity of antigen recognition. EL3 is located between amino acid residues 283 and 300 of SEQ ID NO: 1. EL3 corresponds to the amino acid residues of SEQ ID NO: 2. EL3 is also contained in the polypeptide of SEQ ID NO: 18.

[0055] Each light chain variable domain and heavy chain variable domain (VL and VH, respectively) of a four-chain immunoglobulin has three CDRs, the three CDRs of the light chain variable domain are called VLCDR1, VLCDR2, and VLCDR3, and the three CDRs of the heavy chain variable domain are called VHCDR1, VHCDR2, and VHCDR3. Each light chain variable domain and heavy chain variable domain of a four-chain immunoglobulin has four framework regions (FRs), the four FRs of the light chain variable domain are called LFR1, LFR2, LFR3, and LFR4, and the four FRs of the heavy chain variable domain are called HFR1, HFR2, HFR3, and HFR4.

[0056] By reference to the standard definitions (including reference numbering systems) set forth herein, by reference to the KABAT numbering system, or by applying the IMGT "Collier de Perle" algorithm, one skilled in the art will be able to determine the position of the various regions / domains of an antibody. In this regard, for purposes of defining the sequences of the present invention, it should be noted that the definition of regions / domains may differ between different reference systems. Thus, the regions / domains defined herein include relevant sequences that vary in length or position by approximately + / - 10% within the full-length sequence of the variable domain of an antibody.

[0057] Thus, based on the structure of a four-chain immunoglobulin, an antigen-binding fragment can be defined by comparison with antibody sequences in available databases and the prior art (particularly by comparing the positions of the functional domains in these sequences), noting that the positions of the framework and constant domains of various types of antibodies (particularly IgG, and in particular mammalian IgG) are well defined. Such comparison also involves data related to the three-dimensional structure of the antibody.

[0058] For the purpose of illustrating specific embodiments of the present invention, antigen-binding fragments of the antibodies comprising the variable domain (which includes the CDRs of the antibody) encompass Fv, dsFv, scFv, Fab, Fab', and F(ab')2. An Fv fragment consists of the VL and VH domains of an antibody bound together by hydrophobic interactions; in a dsFv fragment, the VH:VL heterodimer is stabilized by disulfide bonds; in an scFv fragment, the VL and VH domains are linked to each other by a flexible peptide linker, forming a single-chain protein. A Fab fragment is a monomeric fragment obtainable by papain digestion of an antibody; a Fab fragment contains the VH-CH1 fragment of the entire L chain and H chain bound together by disulfide bonds. A F(ab')2 fragment can be produced by pepsin digestion of an antibody below the hinge disulfide bonds; a F(ab')2 fragment contains two Fab' fragments and a portion of the hinge region of an immunoglobulin molecule. A Fab' fragment can be obtained from a F(ab')2 fragment by cleaving the disulfide bonds in the hinge region. F(ab')2 fragments are bivalent, i.e., F(ab')2 fragments contain two antigen-binding sites like natural immunoglobulin molecules; on the other hand, Fv (VHVL dimers constituting the variable portion of Fab), dsFv, scFv, Fab, and Fab' fragments are monovalent, i.e., they contain a single antigen-binding site. These basic antigen-binding fragments of the present invention can be combined together to obtain multivalent antigen-binding fragments, such as bivalent antibodies (diabodies), trivalent antibodies (tribodies), or tetravalent antibodies (tetrabodies). These multivalent antigen-binding fragments are also part of the present invention.

[0059] As used herein, the term "modified antibody" includes "bispecific" antibodies and refers to antibodies that recognize two different antigens by virtue of at least one region having specificity for a first antigen (e.g., a variable region derived from a first antibody) and at least one second region having specificity for a second antigen (e.g., a variable region derived from a second antibody). A bispecific antibody specifically binds to two target antigens and is therefore a multispecific antibody. Multispecific antibodies that recognize two or more different antigens can be produced by recombinant DNA methods, or include, but are not limited to, antibodies produced chemically by any convenient method. Bispecific antibodies include all antibodies or conjugates of antibodies or polymeric forms of antibodies that can recognize two different antigens. Bispecific antibodies include antibodies that have been reduced and recombined to retain their bivalent characteristics, as well as antibodies that have been chemically coupled so that they can have multiple antigen recognition sites for each antigen, such as BiME (bispecific macrophage-enhancing antibody), BiTE (bispecific T cell engager), DART (dual affinity repositioning), DNL (dock-and-lock), DVD-Ig (dual variable domain immunoglobulin), HAS (human serum albumin), and KiH (knob into hole).

[0060] Antigen-binding antibody mimics are organic compounds that specifically bind to antigens but are structurally unrelated to antibodies. Antibody mimics are typically artificial peptides or small proteins with a molar mass of about 3 to 20 kDa. Nucleic acids and small molecules are sometimes also considered as antibody mimics, but do not include artificial antibodies, antibody fragments, and the fusion proteins consisting of them. Compared with antibodies, the common advantages of antibody mimics are solubility, tissue permeability, better stability to heat and enzymes, and relatively low production costs. Antibody mimics are developed as therapeutic and diagnostic agents. Antigen-binding antibody mimics can also be selected from: affibodies, affilins, affinity polymers, affitins, DARPins, and monoclonal antibodies.

[0061] More preferably, the antigen-binding antibody mimetic is selected from Affitin and Anticalin. Affitin is an artificial protein with the ability to selectively bind antigens. Affitin is structurally derived from the DNA binding protein Sac7d found in Sulfolobus acidocaldarius (a microorganism belonging to the Archaebacteria domain). By randomizing the amino acids on the Sac7d binding surface, for example, by generating variants of random substitutions corresponding to 11 residues of the Sac7d binding interface, an Affitin library can be generated and the resulting protein library can be subjected to multiple rounds of ribosome display, and affinity can be directed to various targets (such as peptides, proteins, viruses and bacteria). Affitin is an antibody mimetic and is being developed as a tool in biotechnology. Affitin is also used as a specific inhibitor of various enzymes (Krehenbrink et al., J.mol.Biol., 383:5, 2008). Affitins with the desired binding properties can be easily developed by a skilled person using methods known in the art (particularly methods disclosed in patent application WO2008068637 and the above-cited publications), in particular by using the antigens disclosed herein to generate and screen phage display libraries and / or ribosome display libraries. Anticalins are artificial proteins capable of binding to antigens (proteins or small molecules). Anticalins are antibody mimetics of human lipocalin, a family of natural binding proteins. Anticalinn is about eight times smaller, with a size of about 180 amino acids and a mass of about 20 kDa (Skerra, Febs J., 275: 11, 2008). Anticalin phage display libraries have been generated that can be used to screen and select Anticalins, in particular, with specific binding properties. A skilled person can easily develop anticalins with the desired binding properties using methods known in the art (particularly methods disclosed in EP Patent EP1270725B1, US Patent US8536307B2, Schlehuber and Skerra, Biophys Biophys. Chem., 96: 2-3, 2002 and the publications cited above), in particular by using the antigens disclosed herein to generate and screen phage display libraries and / or ribosome display libraries. Anticalins and affitins can be produced in many expression systems (including bacterial expression systems).Thus, the present invention includes the use of Affitins, Anticalins and other similar antibody mimetics that have the properties of the antibodies described herein, particularly with respect to their ability to bind to CMKLR1, their ability to act as agonists on the binding between RvE1 and CMKLR1, their ability to induce or inhibit the secretion of specific cytokines as described herein, their use in treating or preventing the diseases described herein, all of which are considered mimetics of the present invention.

[0062] As used herein, a "modified antibody" refers to an antibody whose amino acid sequence has been altered by mutation of at least one amino acid residue. Thus, a "modified antibody" encompasses chimeric or humanized antibodies as described herein, and a "modified antibody" may also correspond to a molecule comprising an antibody or antigen-binding fragment thereof, wherein the monoclonal antibody or functional fragment thereof is associated with a functionally different molecule. The modified antibody of the present invention may be a fusion chimeric protein or conjugate produced by any suitable binding form, including covalent binding, grafting, chemical bonding to a chemical group or biological group, or to a molecule (e.g., a PEG polymer or another protective group or molecule suitable for resisting protease cleavage in vivo) to improve the stability and / or half-life of the antibody or functional fragment. Using similar techniques (particularly by chemical coupling or grafting), modified antibodies can be prepared with biologically active molecules, for example, selected from: toxins (particularly Pseudomonas exotoxin A, plant toxins ricin A chain or saporin toxins, particularly therapeutically active ingredients), carriers (particularly protein carriers) suitable for targeting antibodies or functional fragments to specific cells or tissues in the human body, or (particularly when antibody fragments are used) the modified antibodies can be conjugated to a label or linker. PEGylation of antibodies or their functional fragments is a particularly useful embodiment because, particularly for therapeutic applications, PEGylation improves the conditions for delivering the active substance to the host. PEGylation can be site-specific to prevent interference with the recognition site of the antibody or functional fragment, and PEGylation can be performed with high molecular weight PEG. PEGylation can be achieved through free cysteine residues present in the sequence of the antibody or functional fragment or by adding free cysteine residues to the amino sequence of the antibody or functional fragment. According to the present invention, when the term "antibody" is used, it refers to an antibody, an antigen-binding fragment thereof, an antigen-binding antibody mimetic, or a modified antibody.

[0063] A "humanized" form of a non-human (e.g., murine) antibody is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., Fv, Fab, Fab', F(ab')2, or other target binding subsequence of an antibody) that contains minimal sequence derived from a non-human immunoglobulin. Typically, a humanized antibody will substantially contain all of at least one, usually two, variable domains, wherein all CDR regions correspond to the CDR regions and / or humanized forms of CDR regions of a non-human immunoglobulin; all or substantially all FR regions are FR regions of a human immunoglobulin template sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically the constant region of a selected human immunoglobulin template. In a specific embodiment, the present invention relates to an antibody comprising a heavy chain variable region disclosed herein and a light chain variable region disclosed herein, wherein the heavy chain variable region and / or the light chain variable region further comprises a constant region (particularly an Fc region).

[0064] The terms "specifically bind" and "specifically bind to" refer to an antibody, antigen-binding fragment, antigen-binding antibody mimetic, or modified antibody of the invention that binds to a specific antigen with a specific binding affinity of at least 1×10 -6 M, 1×10 -7 M, 1×10 -8 M, 1×10 -9 M, 1×10 -10 M, 1×10 -11 M, 1×10 -12 The ability to bind to CMKLR1 with an affinity of at least 1 M and / or an affinity of at least twice as high as its affinity for a non-specific target (e.g., another protein other than CMKLR1). Affinity can be assessed using various methods known to those skilled in the art. These methods include, but are not limited to, biosensors such as Biacore analysis, Blitz analysis, and Scatchard plots.

[0065] The term "therapeutically effective amount" is used to refer to an amount of any given compound described herein that is sufficient to at least improve the clinical or physiological condition of the patient being treated. The therapeutically effective amount of an antibody, antigen-binding fragment thereof, antigen-binding antibody mimetic, or modified antibody of the invention to be administered depends on considerations such as the disease being treated, the specific mammal being treated, the clinical symptoms of the individual patient, the cause of the disease, the site of delivery of the agent, the method of administration, the schedule of administration, and other factors known to physicians.

[0066] All embodiments disclosed herein with respect to antibodies or antigen-binding fragments thereof are converted, mutatis mutandis, to the macromolecules of the present invention (particularly antigen-binding antibody mimetics and modified antibodies).

[0067] In a first aspect, the present invention relates to an antibody, an antigen-binding fragment thereof, an antigen-binding antibody mimetic or a modified antibody, wherein the antibody, antigen-binding fragment thereof, antigen-binding antibody mimetic or modified antibody specifically binds to CMKLR1 (particularly the EL3 loop of CMKLR1), particularly specifically binds to a polypeptide comprising the amino acid residues of SEQ ID NO: 2, more particularly specifically binds to an epitope located within the EL3 loop of SEQ ID NO: 2 or CMKLR1, the epitope consisting of the sequence of amino acid residues "AMPGS" (SEQ ID NO: 152), and the antibody, antigen-binding fragment thereof, antigen-binding antibody mimetic or modified antibody comprises:

[0068] a heavy chain variable domain comprising HCDR1, HCDR2 and HCDR3;

[0069] - A light chain variable domain comprising LCDR1, LCDR2 and LCDR3.

[0070] In another aspect, the present invention relates to an anti-CMKLR1 compound, wherein the anti-CMKLR1 compound is selected from an antibody or antigen-binding fragment thereof, or an antigen-binding antibody mimetic, or a chimeric antibody or a humanized antibody that specifically binds to CMKLR1, and the compound comprises:

[0071] - an antibody heavy chain variable domain comprising three CDRs (VHCDR1, VHCDR2 and VHCDR3), wherein:

[0072] VHCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 4 or a mutant sequence thereof (in which amino acid residues are substituted); and

[0073] VHCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 6 or a mutant sequence thereof (wherein an amino acid residue is substituted); and

[0074] VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 8 or a mutant sequence thereof (wherein the amino acid residues are substituted such that the amino acid residues at positions 1 and 2 of SEQ ID NO: 8 are L and I or L, respectively); and

[0075] - an antibody light chain variable domain comprising three CDRs (VLCDR1, VLCDR2, and VLCDR3), wherein:

[0076] VLCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 12 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0077] VLCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 14 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0078] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or a mutant sequence thereof (wherein the amino acid residues are substituted);

[0079] wherein the anti-CMKLR1 compound is an RvE1-like agonist of CMKLR1, in particular, the anti-CMKLR1 compound is a pro-resolving factor, in particular targeting the myeloid cell lineage;

[0080] In particular, the anti-CMKLR1 compound is an anti-human CMKLR1 compound.

[0081] In another aspect, the present invention relates to an anti-CMKLR1 compound, wherein the anti-CMKLR1 compound is selected from an antibody or antigen-binding fragment thereof, or an antigen-binding antibody mimetic, or a chimeric antibody or a humanized antibody that specifically binds to CMKLR1, and the anti-CMKLR1 compound comprises an antibody heavy chain variable domain, wherein the antibody heavy chain variable domain comprises VHCDR3, and the VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 8 or a mutant sequence thereof, wherein the amino acid residues in the mutant sequence are substituted such that the amino acid residues at positions 1 and 2 of SEQ ID NO: 8 are L and I or L, respectively; and

[0082] wherein the anti-CMKLR1 compound specifically binds to an epitope located within the third extracellular loop (EL3) of CMKLR1, and in particular, the compound specifically binds to a polypeptide comprising or consisting of amino acid residues of SEQ ID NO: 2 or SEQ ID NO: 152; and

[0083] wherein the anti-CMKLR1 compound is a resolvin E1-like agonist of CMKLR1, in particular, the anti-CMKLR1 compound is a pro-resolvation factor, in particular against myeloid cell lineage; and

[0084] wherein the compound competes with an antibody comprising a heavy chain variable domain corresponding to SEQ ID NO: 9 and a light chain variable domain corresponding to SEQ ID NO: 16 for binding to a polypeptide; more particularly, the compound competes with antibody 2G1 for binding to a polypeptide comprising or consisting of amino acid residues of the sequence SEQ ID NO: 2 or SEQ ID NO: 152, or the polypeptide comprising or consisting of the third loop (EL3) of the extracellular domain of CMKLR1.

[0085] In another aspect, the present invention relates to an anti-CMKLR1 compound, wherein the anti-CMKLR1 compound is selected from an antibody or antigen-binding fragment thereof or an antigen-binding antibody mimetic or a chimeric antibody or a humanized antibody that specifically binds to CMKLR1, and the compound comprises:

[0086] - an antibody heavy chain variable domain comprising three CDRs (i.e., VHCDR1, VHCDR2, and VHCDR3), wherein:

[0087] VHCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 4 or a mutant sequence thereof (in which amino acid residues are substituted); and

[0088] VHCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 6 or a mutant sequence thereof (wherein an amino acid residue is substituted); and

[0089] VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 8 or a mutant sequence thereof (wherein the amino acid residues are substituted such that the amino acid residues at positions 1 and 2 of SEQ ID NO: 8 are L and I or L, respectively); and

[0090] An antibody light chain variable domain comprising three CDRs (i.e., VLCDR1, VLCDR2, and VLCDR3), wherein:

[0091] VLCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 12 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0092] VLCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 14 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0093] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or a mutant sequence thereof (wherein the amino acid residues are substituted);

[0094] wherein the anti-CMKLR1 compound is an RvE1-like agonist of CMKLR1, in particular, the anti-CMKLR1 compound is a pro-resolving factor, in particular targeting the myeloid cell lineage;

[0095] The compound competes for binding to a polypeptide (or antigen) and / or the third extracellular loop of CMKLR1 with an antibody comprising a heavy chain variable domain corresponding to SEQ ID NO: 9 and a light chain variable domain corresponding to SEQ ID NO: 16; more specifically, the compound competes for binding to a polypeptide (or antigen) and / or the third extracellular loop of CMKLR1 with antibody 2G1, wherein the polypeptide (or antigen) comprises or consists of amino acid residues of SEQ ID NO: 2 and / or SEQ ID NO: 152 (i.e., cross-competes for binding to amino acid residues of SEQ ID NO: 2 and / or SEQ ID NO: 152 or the third extracellular loop of CMKLR1). Binding to a polypeptide comprising or consisting of amino acid residues of SEQ ID NO: 2 and / or SEQ ID NO: 152 and / or binding to the third extracellular loop of CMKLR1 can be assessed by the Examples disclosed herein, particularly by analyzing binding affinity by ELISA in Example 9. To determine whether a test antibody can compete for binding to the same antigen or the third loop when the epitope is bound by a 2G1 antibody (or an antigen-binding fragment comprising a heavy chain variable domain corresponding to SEQ ID NO: 9 and a light chain domain corresponding to SEQ ID NO: 16), a cross-blocking assay (e.g., a competition ELISA assay) can be performed. In one exemplary competition ELISA assay, a polypeptide comprising or consisting of the epitope or the third loop can be coated onto the wells of a microtiter plate, pre-incubated with or without a candidate competing antibody, and then a biotinylated 2G1 antibody of the invention is added. Using an avidin-peroxidase conjugate and an appropriate substrate, the amount of labeled anti-2G1 antibody bound to the polypeptide (comprising or consisting of the polypeptide of SEQ ID NO: 2 or SEQ ID NO: 152) or the third loop of CMKLR1 in the wells is measured. The antibody can be labeled with a radioactive or fluorescent label or some other detectable and measurable label. The amount of labeled anti-2G1 antibody bound to the polypeptide or third loop of SEQ ID NO: 2 or SEQ ID NO: 152 is indirectly related to the ability of the candidate competing antibody (test antibody) to compete for binding to the same epitope or the same loop. That is, the greater the affinity of the test antibody for the same epitope, the less labeled 2G1 antibody will bind to the antigen-coated wells. A candidate competing antibody is considered to compete for binding to the same polypeptide or third loop with a 2G1 antibody of the invention if it blocks the binding of the 2G1 antibody by at least 20% (preferably at least 20% to 50%, more preferably at least 50%) compared to a control performed in parallel in the absence of the candidate competing antibody (but with the presence of a known non-competing antibody). It will be appreciated that variations in this assay can be performed to achieve the same quantitative value.

[0096] In another aspect, the present invention relates to an antibody, an antigen-binding fragment thereof, an antigen-binding antibody mimetic or a chimeric antibody or a humanized antibody that specifically binds to CMKLR1 (particularly human CMKLR1), comprising:

[0097] - an antibody heavy chain variable domain comprising three CDRs (i.e., VHCDR1, VHCDR2, and VHCDR3), wherein:

[0098] HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64 or SEQ ID NO: 65; and

[0099] VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71 or SEQ ID NO: 72; and

[0100] VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150 or SEQ ID NO: 151; and / or

[0101] An antibody light chain variable domain comprising three CDRs (i.e., VLCDR1, VLCDR2, and VLCDR3), wherein:

[0102] VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79 or SEQ ID NO: 80; and

[0103] VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87 or SEQ ID NO: 88; and

[0104] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 89.

[0105] In one embodiment, the present invention relates to an antibody, antigen-binding fragment, antigen-binding antibody mimetic or modified antibody that specifically binds to CMKLR1 (particularly human CMKLR1), comprising:

[0106] - an antibody heavy chain variable domain comprising three CDRs (i.e., VHCDR1, VHCDR2, and VHCDR3), wherein:

[0107] VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 62 or SEQ ID NO: 63; and

[0108] VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 70 or SEQ ID NO: 72; and

[0109] VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 8, SEQ ID NO: 144 or SEQ ID NO: 148; and / or

[0110] An antibody light chain variable domain comprising three CDRs (i.e., VLCDR1, VLCDR2, and VLCDR3), wherein:

[0111] VLCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 77; and

[0112] VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 81 or SEQ ID NO: 84; and

[0113] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 89.

[0114] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 4, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 67; and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly, the amino acid sequence shown in SEQ ID NO: 144).

[0115] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 4, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 70, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly, the amino acid sequence shown in SEQ ID NO: 144).

[0116] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 4, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 72, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly, the amino acid sequence shown in SEQ ID NO: 144).

[0117] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 62, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 67, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0118] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 62, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 70, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0119] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 62, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 72, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0120] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 63, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 67, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0121] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 63, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 70, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0122] In a specific embodiment of the present invention, the antibody heavy chain variable domain comprises: VHCDR1 of the amino acid sequence shown in SEQ ID NO: 63, VHCDR2 of the amino acid sequence shown in SEQ ID NO: 72, and VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO: 148 (more particularly VHCDR3 of the amino acid sequence shown in SEQ ID NO: 144).

[0123] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1 with the amino acid sequence shown in SEQ ID NO: 77, VLCDR2 with the amino acid sequence shown in SEQ ID NO: 14, and VLCDR3 with the amino acid sequence shown in SEQ ID NO: 15.

[0124] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1 with the amino acid sequence shown in SEQ ID NO: 77, VLCDR2 with the amino acid sequence shown in SEQ ID NO: 14, and VLCDR3 with the amino acid sequence shown in SEQ ID NO: 89.

[0125] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1:77 of the amino acid sequence shown in SEQ ID NO:1, VLCDR2 of the amino acid sequence shown in SEQ ID NO:81, and VLCDR3 of the amino acid sequence shown in SEQ ID NO:15.

[0126] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1 with the amino acid sequence shown in SEQ ID NO: 77, VLCDR2 with the amino acid sequence shown in SEQ ID NO: 81, and VLCDR3 with the amino acid sequence shown in SEQ ID NO: 89.

[0127] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1 with the amino acid sequence shown in SEQ ID NO: 77, VLCDR2 with the amino acid sequence shown in SEQ ID NO: 84, and VLCDR3 with the amino acid sequence shown in SEQ ID NO: 15.

[0128] In a specific embodiment of the present invention, the antibody light chain variable domain comprises: VLCDR1 with the amino acid sequence shown in SEQ ID NO: 77, VLCDR2 with the amino acid sequence shown in SEQ ID NO: 84, and VLCDR3 with the amino acid sequence shown in SEQ ID NO: 89.

[0129] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody is a humanized antibody of a murine antibody, in particular, at least one framework region (FR1 and / or FR2 and / or FR3 and / or FR4) of the antibody light chain variable domain and / or the antibody heavy chain variable domain is derived from a human chain framework region, and the light chain and / or heavy chain further comprises any combination of CDRs or wild-type CDRs described herein.

[0130] Starting from a variety of choices of the amino acid sequence of the third CDR of the heavy chain, wild-type antibodies have been defined and synthesized, referred to herein as 2G1 and 2G4 (having the same variable region but different heavy chain constant regions). Therefore, several putative antibody sequences have been produced and synthesized and their binding and biological properties have been tested. In a variety of newly synthesized antibodies, it has been determined that the presence of amino acid residues "5'RLIY 3'" or "5'RLLY 3'" sequences in the third CDR of the heavy chain variable domain (according to IMGT) is highly correlated with the agonist properties of the antibody to RvE1-CMKLR1 interactions. Antibodies with different amino acid residue sequences (e.g., "5'RIIY 3'"; "5'RILY 3'") have relatively poor agonist abilities to bind to RvE1 and CMKLR1.

[0131] Therefore, the synthetic wild-type anti-CMKLR1 antibodies (referred to herein as 2G1 and 4G1) are chimeric antibodies having a murine variable region and a human constant region (human IgG1 for 2G1 or human IgG4 for 2G4). 2G1 has the following amino acid sequence in its variable sequence:

[0132] Heavy chain:

[0133] metdtllllwvllllwvpgstgdeVQLVASGGGLVQPGGSLKLSCAAS GFTFSSYG MSWVRQTPDRRLELVAT INRYGGST YYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYC PRLIYYGNEGDS WGQGTTLTVSS

[0134] The signal peptide is indicated in lowercase letters, and the CDRs are shown in bold (according to Kabat) or underlined (according to IMGT). The full sequence of the heavy chain corresponds to SEQ ID NO: 10 with a signal peptide, while SEQ ID NO: 9 corresponds to the heavy chain without a signal peptide. The CDR1, CDR2, and CDR3 according to Kabat correspond to SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 144, respectively. The CDR1, CDR2, and CDR3 according to IMGT correspond to SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7, respectively.

[0135] The CDR3 can be modified by substituting the amino acid residue ILE at position 2 (second according to Kabat numbering) with the amino acid residue LEU, resulting in the heavy chain of the sequence:

[0136] metdtllllwvllllwvpgstgdeVQLVASGGGLVQPGGSLKLSCAAS GFTFSSYG MSWVRQTPDRRLELVAT INRYGGST YYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYC PRLLYYGNEGDS WGQGTTLTVSS

[0137] Light chain:

[0138] metdtlllwvllllwvpgstgdQIVLTQSPAIMSASPGEKVTMTCSAS SSVSF MHWYQQKSGTSPKRWIY DTT KLTSGVPARFSGSGSGTFYSLTISSMEAEDAATYYC QQWNSKPPLT FGPGTKLELKR

[0139] The signal peptide is in lowercase, and the CDRs are in bold (according to Kabat) or underlined (according to IMGT). The full sequence of the light chain corresponds to SEQ ID NO: 17 with a signal peptide, while SEQ ID NO: 16 corresponds to the heavy chain without a signal peptide. The CDR1, CDR2, and CDR3 according to Kabat correspond to SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 15, respectively; the CDR1, CDR2, and CDR3 according to IMGT correspond to SEQ ID NO: 11, SEQ ID NO: 13, and SEQ ID NO: 15, respectively.

[0140] As is known in the art, taking the chimeric antibodies of the present invention as an example, the heavy chain variable domain and the light chain variable domain each comprise three CDRs (CDR1, CDR2, and CDR3, respectively, from the 5' end to the 3' end) and four framework regions (FR1, FR2, FR3, and FR4, respectively, from the 5' end to the 3' end). Humanization of murine antibodies can include humanizing at least one framework region within the light chain variable region or the heavy chain variable region, or both. In one embodiment, several framework regions can be humanized, particularly within the heavy chain variable region and within the light chain variable region. Wild-type CDRs can be conserved, but CDRs can also be replaced by CDRs described herein. Therefore, when the framework regions are humanized, the anti-CKLMR1 compounds according to the present invention may comprise at least one, at least two, at least three, at least four, at least five, or six wild-type CDRs. In other words, the anti-CMKLR1 compound is a humanized form of the parent chimeric antibody 2G1, wherein at least one framework region is humanized, and in particular, at least one framework region and at least one CDR are humanized. In one embodiment of the present invention, the variable region of the antibody can be associated with an antibody constant region, such as the constant regions set forth in SEQ ID NO: 134 (encoded by the nucleotide sequence of SEQ ID NO: 133), SEQ ID NO: 136 (encoded by the nucleotide sequence of SEQ ID NO: 135), SEQ ID NO: 138 (encoded by the nucleotide sequence of SEQ ID NO: 137), SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO: 141. This combination is exemplified by the light chain of SEQ ID NO: 142 and the heavy chain of SEQ ID NO: 143.

[0141] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises at least one humanized framework region within its antibody heavy chain variable domain, wherein:

[0142] The antibody heavy chain variable domain comprises four framework regions: HFR1, HFR2, HFR3 and HFR4, wherein:

[0143] HFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92 or SEQ ID NO: 93; and / or

[0144] HFR2 comprises or consists of the amino acid sequence of SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99 or SEQ ID NO: 100; and / or

[0145] HFR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105 or SEQ ID NO: 106; and / or

[0146] HFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 107 or SEQ ID NO: 108.

[0147] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises at least one humanized framework region within its antibody light chain variable domain, wherein:

[0148] The antibody light chain variable domain comprises four framework regions: LFR1, LFR2, LFR3 and LFR4, wherein:

[0149] LFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111 or SEQ ID NO: 112; and / or

[0150] LFR2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116 or SEQ ID NO: 117; and / or

[0151] LFR3 comprises or consists of the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122 or SEQ ID NO: 132; and / or

[0152] LFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 123 or SEQ ID NO: 124.

[0153] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises at least one humanized framework region in its antibody light chain variable region and its antibody heavy chain variable region, wherein:

[0154] The antibody heavy chain variable domain comprises four framework regions: HFR1, HFR2, HFR3 and HFR4, wherein:

[0155] HFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92 or SEQ ID NO: 93; and / or

[0156] HFR2 comprises or consists of the amino acid sequence of SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99 or SEQ ID NO: 100; and / or

[0157] HFR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105 or SEQ ID NO: 106; and / or

[0158] HFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 107 or SEQ ID NO: 108; and

[0159] The antibody light chain variable domain comprises four framework regions: LFR1, LFR2, LFR3 and LFR4, wherein:

[0160] LFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111 or SEQ ID NO: 112; and / or

[0161] LFR2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116 or SEQ ID NO: 117; and / or

[0162] LFR3 comprises or consists of the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122 or SEQ ID NO: 132; and / or

[0163] LFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 123 or SEQ ID NO: 124.

[0164] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0165] - a heavy chain variable region comprising HFR1 of SEQ ID NO: 91, HFR2 of SEQ ID NO: 95, HFR3 of SEQ ID NO: 102, and HFR4 of SEQ ID NO: 108.

[0166] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0167] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0168] - a heavy chain variable region comprising HFR1 of SEQ ID NO: 92, HFR2 of SEQ ID NO: 96, HFR3 of SEQ ID NO: 103, and HFR4 of SEQ ID NO: 108.

[0169] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0170] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0171] - a heavy chain variable region comprising HFR1 of SEQ ID NO: 92, HFR2 of SEQ ID NO: 98, HFR3 of SEQ ID NO: 106, and HFR4 of SEQ ID NO: 108.

[0172] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0173] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0174] - a heavy chain variable region comprising HFR1 of SEQ ID NO: 93, HFR2 of SEQ ID NO: 96, HFR3 of SEQ ID NO: 104, and HFR4 of SEQ ID NO: 108.

[0175] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0176] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0177] - a light chain variable region comprising LFR1 of SEQ ID NO: 111, LFR2 of SEQ ID NO: 115, LFR3 of SEQ ID NO: 120, and LFR4 of SEQ ID NO: 124.

[0178] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0179] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0180] - a light chain variable region comprising LFR1 of SEQ ID NO: 110, LFR2 of SEQ ID NO: 114, LFR3 of SEQ ID NO: 119, and LFR4 of SEQ ID NO: 124.

[0181] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0182] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0183] - a light chain variable region comprising LFR1 of SEQ ID NO: 110, LFR2 of SEQ ID NO: 114, LFR3 of SEQ ID NO: 122, and LFR4 of SEQ ID NO: 124.

[0184] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0185] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0186] - a light chain variable region comprising LFR1 of SEQ ID NO: 112, LFR2 of SEQ ID NO: 116, LFR3 of SEQ ID NO: 121 and LFR4 of SEQ ID NO: 124.

[0187] Such an antibody may comprise the CDRs of the parent chimeric antibody (i.e., HCDR1 of SEQ ID NO: 4; HCDR2 of SEQ ID NO: 6; HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; LCDR1 of SEQ ID NO: 12; LCDR2 of SEQ ID NO: 14; and LCDR3 of SEQ ID NO: 15).

[0188] In one embodiment, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a humanized antibody light chain variable domain and a humanized antibody heavy chain variable domain, wherein:

[0189] -Antibody heavy chain variable domain comprising:

[0190] VFICDR1 having the amino acid sequence set forth in SEQ ID NO: 4; VFICDR2 having the amino acid sequence set forth in SEQ ID NO: 67; and VFICDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0191] VFICDR1 having the amino acid sequence set forth in SEQ ID NO: 4; VFICDR2 having the amino acid sequence set forth in SEQ ID NO: 70; and VFICDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0192] VFICDR1 having the amino acid sequence set forth in SEQ ID NO: 4; VFICDR2 having the amino acid sequence set forth in SEQ ID NO: 72; and VFICDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0193] VFICDR1 having the amino acid sequence set forth in SEQ ID NO: 62; VFICDR2 having the amino acid sequence set forth in SEQ ID NO: 67; and VFICDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0194] VHCDR1 having the amino acid sequence set forth in SEQ ID NO: 62; VHCDR2 having the amino acid sequence set forth in SEQ ID NO: 70; and VHCDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0195] VHCDR1 having the amino acid sequence set forth in SEQ ID NO: 62; VHCDR2 having the amino acid sequence set forth in SEQ ID NO: 72; and VHCDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0196] VHCDR1 having the amino acid sequence set forth in SEQ ID NO: 63; VHCDR2 having the amino acid sequence set forth in SEQ ID NO: 67; and VHCDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0197] VHCDR1 having the amino acid sequence set forth in SEQ ID NO: 63; VHCDR2 having the amino acid sequence set forth in SEQ ID NO: 70; and VHCDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; or

[0198] VHCDR1 having the amino acid sequence set forth in SEQ ID NO: 63; VHCDR2 having the amino acid sequence set forth in SEQ ID NO: 72; and VHCDR3 having the amino acid sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148; and

[0199] -Antibody light chain variable domain comprising:

[0200] VLCDR1 having the amino acid sequence set forth in SEQ ID NO: 77; VLCDR2 having the amino acid sequence set forth in SEQ ID NO: 14; and VLCDR3 having the amino acid sequence set forth in SEQ ID NO: 15; or

[0201] VLCDR1 having the amino acid sequence set forth in SEQ ID NO: 77; VLCDR2 having the amino acid sequence set forth in SEQ ID NO: 14; and VLCDR3 having the amino acid sequence set forth in SEQ ID NO: 89; or

[0202] VLCDR1 having the amino acid sequence set forth in SEQ ID NO: 77; VLCDR2 having the amino acid sequence set forth in SEQ ID NO: 81; and VLCDR3 having the amino acid sequence set forth in SEQ ID NO: 15; or

[0203] VLCDR1 having the amino acid sequence set forth in SEQ ID NO: 77; VLCDR2 having the amino acid sequence set forth in SEQ ID NO: 81; and VLCDR3 having the amino acid sequence set forth in SEQ ID NO: 89;

[0204] VLCDR1 having the amino acid sequence set forth in SEQ ID NO: 77; VLCDR2 having the amino acid sequence set forth in SEQ ID NO: 84; and VLCDR3 having the amino acid sequence set forth in SEQ ID NO: 15; or

[0205] VLCDR1 having the amino acid sequence shown in SEQ ID NO: 77; VLCDR2 having the amino acid sequence shown in SEQ ID NO: 84; and VLCDR3 having the amino acid sequence shown in SEQ ID NO: 89.

[0206] It should be noted that combinations of these CDRs (VHCDRs and VLCDRs) can be combined with the humanized framework regions described herein, in particular with the following framework regions:

[0207] - HFR1 of SEQ ID NO: 91, HFR2 of SEQ ID NO: 95, HFR3 of SEQ ID NO: 102, and HFR4 of SEQ ID NO: 108; or

[0208] - HFR1 of SEQ ID NO: 92, HFR2 of SEQ ID NO: 96, HFR3 of SEQ ID NO: 103 and HFR4 of SEQ ID NO: 108; or

[0209] - HFR1 of SEQ ID NO: 92, HFR2 of SEQ ID NO: 98, HFR3 of SEQ ID NO: 106 and HFR4 of SEQ ID NO: 108; or

[0210] - HFR1 of SEQ ID NO: 93, HFR2 of SEQ ID NO: 96, HFR3 of SEQ ID NO: 104, and HFR4 of SEQ ID NO: 108; and

[0211] - a light chain variable region comprising LFR1 of SEQ ID NO: 111, LFR2 of SEQ ID NO: 115, LFR3 of SEQ ID NO: 120, and LFR4 of SEQ ID NO: 124; or

[0212] - a light chain variable region comprising LFR1 of SEQ ID NO: 110, LFR2 of SEQ ID NO: 114, LFR3 of SEQ ID NO: 119, and LFR4 of SEQ ID NO: 124; or

[0213] - a light chain variable region comprising LFR1 of SEQ ID NO: 110; LFR2 of SEQ ID NO: 114, LFR3 of SEQ ID NO: 122, and LFR4 of SEQ ID NO: 124; or

[0214] - a light chain variable region comprising LFR1 of SEQ ID NO: 112, LFR2 of SEQ ID NO: 116, LFR3 of SEQ ID NO: 121 and LFR4 of SEQ ID NO: 124.

[0215] In a specific embodiment of the invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42; in particular, the heavy chain variable domain comprises or consists of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, NO: 23, SEQ ID NO: 28: SEQ ID NO, 29, SEQ ID NO: 30, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39.

[0216] In a specific embodiment of the invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; in particular, the heavy chain variable domain comprises or consists of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59 or SEQ ID NO: 60.

[0217] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0218] a heavy chain variable domain comprising HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148, HFR1 of SEQ ID NO: 90, HFR2 of SEQ ID NO: 94, HFR3 of SEQ ID NO: 101, and HFR4 of SEQ ID NO: 107; and

[0219] - a light chain variable domain comprising LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, LCDR3 of SEQ ID NO: 15, LFR1 of SEQ ID NO: 109, LFR2 of SEQ ID NO: 113, LFR3 of SEQ ID NO: 118 and LFR4 of SEQ ID NO: 123.

[0220] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises the heavy chain variable domain of SEQ ID NO: 10 and the light chain variable domain of SEQ ID NO: 17.

[0221] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0222] - a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42; in particular, the heavy chain variable domain comprises or consists of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 39;

[0223] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; in particular, a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59 or SEQ ID NO: 60.

[0224] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein HFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 19 (E / Q VQLV A / E / Q SG G / A / SGL / EV / LQ / KP / K PG G / ASL / VK / R / VL / V SC A / K AS).

[0225] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein HFR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 125 (WVR Q / A TP D / GR / KR / G / Q LELVA).

[0226] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein HFR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 126 (RF / VT / VIS / TRDN A / SK / T / VN / S TLY L / MQ / EM / L / I SSL K / RS / A EDTA M / VYYCPR).

[0227] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein HFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 127 (WGQGT T / LL / V TVSS).

[0228] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein LFR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 128 (Q / A / EIV / Q LTQSP A / S / DI / S / F / TM / L / QSA / S / LS / VP / V / TG / PE / D / KK / RV / A TM / I / L TC).

[0229] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein LFR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 129 (WYQQK S / PG / DT / KS / APK / RRWIY).

[0230] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein LFR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 130 (GV / IP A / S RFSGSGSGT F / DYS / T LTI S / NSM / LE / QA / P ED A / FAT / V YYC).

[0231] In a specific embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises a heavy chain variable domain, wherein LFR4 comprises or consists of the amino acid sequence shown in SEQ ID NO: 131 (FGP / GGTKL / VEL / IKR).

[0232] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0233] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 20; and

[0234] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0235] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0236] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 21; and

[0237] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0238] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0239] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 22; and

[0240] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0241] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0242] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 23; and

[0243] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0244] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0245] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 24; and

[0246] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0247] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0248] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 25; and

[0249] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0250] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0251] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 26; and

[0252] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0253] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0254] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 27; and

[0255] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0256] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0257] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 28; and

[0258] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0259] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0260] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 29; and

[0261] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0262] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0263] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 30; and

[0264] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0265] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0266] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 31; and

[0267] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0268] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0269] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 32; and

[0270] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0271] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0272] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 33; and

[0273] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0274] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0275] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 34; and

[0276] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0277] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0278] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 35; and

[0279] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0280] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0281] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 36; and

[0282] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0283] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0284] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 37; and

[0285] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0286] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0287] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 38; and

[0288] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0289] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0290] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 39; and

[0291] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0292] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0293] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 40; and

[0294] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0295] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0296] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 41; and

[0297] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0298] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0299] - a heavy chain variable domain comprising or consisting of the amino acid sequence shown in SEQ ID NO: 42; and

[0300] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61.

[0301] The present invention relates to any of the above compounds of the present invention for use in preventing and / or treating diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome ( syndrome), celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders, dry eye syndrome; cancer diseases, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers (carcinoma), in particular mammary carcinoma or colon carcinoma, or lung or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1. In one embodiment, the present invention relates to any of the above-mentioned compounds of the present invention for use in the prevention and / or treatment of diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type 1 diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders and dry eye syndrome. In another embodiment of the present invention, the present invention relates to any of the above-mentioned compounds of the present invention for use in the prevention and / or treatment of cancer, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer, or lung or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1.

[0302] In one embodiment, the present invention relates to an anti-CMKLR1 compound selected from an antibody that specifically binds to CMKLR1 or an antigen-binding fragment thereof or an antibody that mimics or modifies an antigen-binding antibody, the compound comprising:

[0303] An antibody heavy chain variable domain comprising three CDRs (i.e., VHCDR1, VHCDR2, and VHCDR3), wherein:

[0304] HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64 or SEQ ID NO: 65; and

[0305] VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71 or SEQ ID NO: 72; and

[0306] VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150 or SEQ ID NO: 151; and / or

[0307] An antibody light chain variable domain comprising three CDRs (i.e., VLCDR1, VLCDR2, and VLCDR3), wherein:

[0308] VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79 or SEQ ID NO: 80; and

[0309] VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87 or SEQ ID NO: 88; and

[0310] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 89.

[0311] The anti-CMKLR1 compounds are used to prevent and / or treat diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the following group: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type 1 diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders, dry eye syndrome; cancer diseases, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer, or lung or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1. In one embodiment, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of diseases in which resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders and dry eye syndrome. In another embodiment of the present invention, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of cancer, in particular solid cancers and liquid cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1.

[0312] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0313] a heavy chain variable domain comprising HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148, HFR1 of SEQ ID NO: 90, HFR2 of SEQ ID NO: 94, HFR3 of SEQ ID NO: 101, and HFR4 of SEQ ID NO: 107; and

[0314] - a light chain variable domain comprising LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, LCDR3 of SEQ ID NO: 15, LFR1 of SEQ ID NO: 109, LFR2 of SEQ ID NO: 113, LFR3 of SEQ ID NO: 118, and LFR4 of SEQ ID NO: 123;

[0315] The antibodies or antigen-binding fragments thereof or antigen-binding antibody mimetics or modified antibodies are used to treat and / or prevent diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type 1 diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders, dry eye syndrome; cancer diseases, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer, or lung or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1. In one embodiment, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of diseases in which resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders and dry eye syndrome. In another embodiment of the present invention, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of cancer, in particular solid cancers and liquid cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1.

[0316] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises the heavy chain variable domain of SEQ ID NO: 10 and the light chain variable domain of SEQ ID NO: 17;

[0317] The antibodies or antigen-binding fragments thereof or antigen-binding antibody mimetics or modified antibodies are used to prevent and / or treat diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the following group: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type 1 diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders, dry eye syndrome; cancer diseases, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer, or lung or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1. In one embodiment, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of diseases in which resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders and dry eye syndrome. In another embodiment of the present invention, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of cancer, in particular solid cancers and liquid cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1.

[0318] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0319] a heavy chain variable domain comprising the HCDR1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 6, the HCDR3 of SEQ ID NO: 8 or SEQ ID NO: 144 or SEQ ID NO: 148, and:

[0320] HFR1 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 91, SEQ ID NO: 92 or SEQ ID NO: 93; and

[0321] HFR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99 or SEQ ID NO: 100; and

[0322] HFR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105 or SEQ ID NO: 106; and

[0323] HFR4 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 108; and

[0324] - a light chain variable domain comprising LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, LCDR3 of SEQ ID NO: 15, and:

[0325] LFR1 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 110, SEQ ID NO: 111 or SEQ ID NO: 112; and

[0326] LFR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116 or SEQ ID NO: 117; and

[0327] LFR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122 or SEQ ID NO: 132; and

[0328] LFR4 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 124.

[0329] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody comprises:

[0330] - a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42; in particular, the heavy chain variable domain comprises or consists of the amino acid sequence of SEQ ID No: 20, SEQ ID No: 21, SEQ ID No: 22, SEQ ID No: 23, SEQ ID No: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: No: 36, SEQ ID No: 37, SEQ ID No: 38 or SEQ ID No: 39;

[0331] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; in particular, a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59 or SEQ ID NO: 60;

[0332] The antibody or its antigen-binding fragment or antigen-binding antibody mimetic or modified antibody is used to prevent and / or treat diseases in which the resolution of inflammation is delayed or impaired and / or diseases selected from the following group: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders, dry eye syndrome; cancer diseases, in particular solid cancers and liquid cancers, metastatic cancers, in particular epithelial cancers, in particular breast epithelial cancer, lung cancer or colon epithelial cancer or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1. In one embodiment, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of diseases in which resolution of inflammation is delayed or impaired and / or diseases selected from the group consisting of: inflammatory diseases, in particular acute inflammatory diseases, chronic inflammatory diseases, such as asthma, keratoconjunctivitis, periodontal disease, eczema, inflammatory bowel disease, in particular Crohn's disease or colitis (in particular ulcerative colitis or idiopathic colitis); autoimmune diseases, such as diabetes (in particular type I diabetes), psoriasis, lupus, rheumatoid arthritis, multiple sclerosis, Sjögren's syndrome, celiac disease, vasculitis, myasthenia gravis; infectious diseases, such as sepsis, peritonitis; degenerative diseases; wound healing disorders and dry eye syndrome. In another embodiment of the present invention, the present invention relates to any of the above compounds of the present invention for use in the prevention and / or treatment of cancer, in particular solid cancers and liquid cancers, in particular epithelial cancers, in particular breast epithelial cancer or colon epithelial cancer or medullary cancer, in particular leukemia, in particular cancers in which cancer cells express CMKLR1 or the tumor microenvironment is invaded by cells expressing or overexpressing CMKLR1.

[0333] In one embodiment of the present invention, the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody has:

[0334] - a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42; in particular, the heavy chain variable domain comprises or consists of the amino acid sequence of SEQ ID No: 20, SEQ ID No: 21, SEQ ID No: 22, SEQ ID No: 23, SEQ ID No: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: No: 36, SEQ ID No: 37, SEQ ID No: 38 or SEQ ID No: 39; and

[0335] - a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; in particular, a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61; NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59 or SEQ ID NO: 60; the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody is used to treat colitis or Crohn's disease in a subject who is refractory to corticosteroid and / or immunosuppressive therapy.

[0336] In another aspect, the present invention relates to an anti-CMKLR1 compound as described herein that enhances anti-inflammatory cytokines, particularly I10, secreted by macrophages. In another aspect, the present invention relates to an anti-CMKLR1 compound as described herein that enhances anti-inflammatory cytokines, particularly CCL17, secreted by macrophages. In another aspect, the present invention relates to an anti-CMKLR1 compound as described herein that enhances anti-inflammatory cytokines, particularly I10 and / or CCL17, secreted by macrophages. The present invention also relates to an anti-CMKLR1 compound as described herein that inhibits pro-inflammatory cytokines, particularly IL-12, secreted by macrophages. The secretion of cytokines can be assessed by methods known in the art or by the methods disclosed in the Examples of the present invention.

[0337] In another aspect, the present invention relates to an anti-CMKLR1 compound as described herein that inhibits the proliferation and / or activation of dendritic cells (particularly human dendritic cells). The proliferation and / or activation of dendritic cells can be assessed by the methods disclosed in the examples of the present invention.

[0338] In another aspect, the present invention relates to a composition comprising an anti-CMKLR1 compound as described herein, in particular a pharmaceutical composition comprising an anti-CMKLR1 compound of the present invention and an additional therapeutic agent or a pharmaceutically acceptable carrier. In a specific embodiment, the present invention relates to a composition comprising an anti-CMKLR1 compound of the present invention and a therapeutic agent selected from an immunomodulator, an immune checkpoint blocker, an immune checkpoint activator, an antibody (in particular an anti-CD137 antibody (which targets 4-1BB, a member of the TNF receptor superfamily also known as CD137)) or an anti-SIRPα antibody (P84 - anti-mouse SIRPα from Merck Millipore).

[0339] In another aspect, the present invention relates to a compound combination comprising the anti-CMKLR1 compound described herein, in particular a pharmaceutical composition comprising an anti-CMKLR1 compound of the present invention and an anti-PD1 or anti-PDL1 compound (in particular an anti-PD1 compound); in particular, such a compound is selected from antibodies, antigen-binding antibody fragments, antigen-binding antibody mimetics, small molecules (such as aptamers or peptides), modified antibodies, such as but not limited to humanized antibodies or chimeric antibodies that can bind to PD1 or PDL1.

[0340] In another aspect, the present invention relates to a compound combination comprising an anti-CMKLR1 compound as described herein, in particular a pharmaceutical composition comprising an anti-CMKLR1 compound of the present invention and an anti-SIRPα compound; in particular, such a compound is selected from antibodies, antigen-binding antibody fragments, antigen-binding antibody mimetics, small molecules (such as aptamers or peptides), modified antibodies such as but not limited to humanized antibodies or chimeric antibodies that can bind to SIRPα (in particular human SIRPα).

[0341] In another aspect, the present invention relates to the therapeutic use of the anti-CMKLR1 compounds of the present invention, in particular for inducing and / or enhancing the resolution of inflammation, in particular when said resolution is delayed or impaired, with a view to the treatment of diseases in which the prolongation of inflammation is pathological or the duration of the resolution of inflammation is pathological.

[0342] In one embodiment of the present invention, the anti-CKLMR1 compound binds to CMKLR1 with an affinity (KD value) of at least 10E-8 M (more preferably at least 10E-9 M). Specific binding of the antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic or modified antibody of the present invention to CMKLR1 (or a region of CMKLR1 comprising the third extracellular loop, comprising the amino acid sequence set forth in SEQ ID NOs: 2 and 18) indicates that the antibody exhibits significant affinity for CMKLR1. "Significant affinity" includes an affinity of about 10E-8 M. -8Preferably, when the binding affinity is 10 -8 M to 10 -12 M (optionally 10 -9 M to 10 -10 M, especially at least 10 -9 M), then the binding is considered specific. Whether the binding domain specifically reacts or binds to the target can be easily detected by comparing the reaction of the binding domain with the target protein or antigen and the reaction of the binding domain with proteins or antigens other than the target protein. Such antibodies of the present invention specifically bind to CMKLR1 and have an agonist effect on the interaction between RvE1 and CMKLR1. Methods for determining antibody specificity and affinity by competitive inhibition are known in the art (see, for example, Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1998); Colligan et al., Current Protocols in Immunology, Green Publishing Assoc., NY (1992; 1993); Meth Muller, Enzym., 92: 589-601 (1983)). These methods include, but are not limited to, Biacore analysis, Blitz analysis, flow cytometry, and ELISA assays.

[0343] In one embodiment of the present invention, the anti-CMKLR1 compound specifically binds to an epitope located within the third extracellular loop of CMKLR1, particularly an epitope within the amino acid residue sequence set forth in SEQ ID NO: 2 or SEQ ID NO: 18 (particularly SEQ ID NO: 2). Anti-CMKLR1 compounds that bind to this specific region of CMKLR1 may exhibit agonist properties for CMKLR1, thereby mimicking the binding of RvE1 to CMKLR1.

[0344] In another aspect, the present invention relates to an anti-CMKLR1 antibody or an antigen-binding fragment or antigen-binding antibody mimetic or modified antibody as described above, which has agonist capacity for the interaction between RvE1 and CMKLR1 for use as a medicament.

[0345] In another aspect, the present invention relates to an anti-CMKLR1 antibody, or an antigen-binding fragment thereof, or an antigen-binding antibody mimetic, or a modified antibody as described above, which has the ability to induce activation of Akt and / or Erk proteins in vitro and / or in vivo. Activation of these proteins can be assessed by the methods described in the Examples of the present invention. In particular, the anti-CMKLR1 antibody, or an antigen-binding fragment thereof, or an antigen-binding antibody mimetic, or a modified antibody thereof, has the ability to activate Akt and / or Erk proteins, or both, in macrophages, particularly human macrophages.

[0346] The present invention also relates to a method for treating a subject in need thereof, comprising administering to the subject an effective amount of an anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic as described above, wherein the anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic has agonist ability for the interaction between RvE1 and CMKLR1, or in other words, the anti-CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic is an RvE1 agonist-like factor or regulator.

[0347] Altering the polarization of macrophages to favor anti-inflammatory cells may be useful in many pathologies or situations. As described above, such alterations are particularly useful for diseases selected from the group consisting of inflammatory diseases, including but not limited to acute inflammatory diseases and chronic inflammatory diseases, inflammatory bowel disease, Crohn's disease, asthma, keratoconjunctivitis, periodontal disease, eczema, colitis, especially ulcerative colitis or idiopathic colitis, diabetes (especially type I diabetes), peritonitis, psoriasis, epithelial cancer (especially breast or colon), cancer, metastatic cancer, lung cancer, degenerative diseases, infectious diseases (especially sepsis), and autoimmune diseases.

[0348] The present invention also relates to the use of the anti-CMKLR1 antibody or its antigen-binding fragment or antigen-binding antibody mimetic as described above in the preparation of a medicament, wherein the anti-CMKLR1 antibody or its antigen-binding fragment or antigen-binding antibody mimetic has agonist ability on the interaction between RvE1 and CMKLR1.

[0349] In another aspect, the present invention relates to an anti-CMKLR1 antibody or an antigen-binding fragment or antigen-binding antibody mimetic or modified antibody thereof, such as but not limited to a humanized or chimeric antibody as described above, for use in treating chronic inflammatory diseases, in particular for treating chronic colitis.

[0350] In another aspect, the present invention relates to an anti-CMKLR1 antibody or an antigen-binding fragment or antigen-binding antibody mimetic as described above, which has agonist activity on the interaction between RvE1 and CMKLR1, for treating the delay or disruption of the resolution of inflammatory conditions (especially inflammatory diseases in which resolution is delayed or disrupted) and / or treating or preventing a disease selected from the following group: inflammatory diseases, including but not limited to acute inflammatory diseases and chronic inflammatory diseases, inflammatory bowel disease, Crohn's disease, asthma, keratoconjunctivitis, periodontal disease, eczema, colitis, especially ulcerative colitis or idiopathic colitis, diabetes (especially type 1 diabetes), peritonitis, psoriasis, epithelial cancer (especially breast epithelial cancer or colon epithelial cancer), cancer, degenerative diseases, infectious diseases (especially sepsis), autoimmune diseases.

[0351] As described herein, when the resolution of inflammation is delayed or destroyed compared to normal resolution (i.e., the resolution that occurs in patients experiencing normal resolution after an inflammatory event), "delay or destruction of the resolution of an inflammatory condition" occurs. Regression delay or defects may result in increased infiltration of granulocytes at the inflammatory site. Therefore, regression delay or defects can be assessed by quantifying the granulocytes at the inflammatory site. The number of granulocytes can be measured by, for example, histology, cell counting or indirect biochemical techniques, such as by enzyme-linked immunosorbent assay (ELISA) to elastase or by PCR to granulocyte receptor 1 for molecular quantification. Regression delay or defects can also be assessed by measuring the delay of granulocyte apoptosis, such as by cytological methods using specific antibodies for annexin 5. Defects or delays in the resolution of inflammation can also be determined by quantitatively assessing the synthesis of proinflammatory cytokines (e.g., TNF-α, IL-8 or IL-12) and anti-inflammatory cytokines (e.g., IL-10). The secretion of cytokines can be assessed by ELISA or PCR. Defects or delays in inflammatory resolution can also be determined by assessing the activation of transcription factors involved in the synthesis of inflammatory cytokines (e.g., NF-κB), for example, by nuclear translocation or Western blot, and / or by quantifying the level of IκB degradation. Defects or delays in inflammatory resolution can also be determined by mass spectrometry or quantitative enzyme-linked immunosorbent assays for pro-resolving mediators (e.g., lipoxins, resolvins, protectins, or maresins) or their precursors (e.g., 17-HDOHE or 14-HDOHE). Defects or delays in resolution can result from deficient synthesis of one or more of these mediators. Defects or delays in resolution can also be determined when expression of receptors for resolving molecules is reduced. These receptors can be selected from ALX, CMK1R1, GPR32, or GPR18. Alternatively or additionally, receptors that are internalized into the cytoplasm for processing can also be assessed. Alternatively or additionally, expression of certain receptors for inflammatory cytokines or lipids can also be assessed; overexpression compared to normal levels indicates a delay or defect in inflammatory resolution. These conditions can be measured by histology, cytology, or PCR. Defective resolution can also result in reduced or inhibited conversion of M1 macrophages to M2 macrophages, with impaired phagocytosis or efferocytosis of the same cells. Therefore, as exemplified in the present examples, delayed or defective resolution can be assessed by analyzing the conversion of M1 to M2 macrophages under specific conditions compared to normal conditions.

[0352] According to one embodiment, anti-CMKLR1 compounds can be used to treat an individual suffering from a cancer selected from the group consisting of breast cancer (particularly breast epithelial cancer), melanoma, colon cancer (particularly colon epithelial cancer), leukemia (particularly acute myeloid leukemia), particularly when the cancer cells overexpress CMKLR1.

[0353] In one embodiment, the present invention relates to an anti-human CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic or a modified antibody as described above, for use as described above; wherein the anti-human CMKLR1 antibody or an antigen-binding fragment thereof or an antigen-binding antibody mimetic or a modified antibody of the present invention is administered to a patient with a tumor that is CMKLR1 positive.

[0354] The antibodies or antigen-binding fragments thereof of the present invention can be administered to a subject by various suitable routes, such as intravenous (IV), subcutaneous (SC), or intramuscular (IM). The anti-CMKLR1 compound can be administered alone or in combination with another therapeutic agent (e.g., a second human monoclonal antibody or antigen-binding fragment thereof). In another example, the antibody is administered together with another agent (e.g., an immunosuppressant, an erythropoiesis-stimulating agent (ESA)) and a therapeutic cell composition, etc. In one embodiment, the present invention relates to an anti-CMKLR1 compound or antigen-binding fragment thereof or an antigen-binding antibody mimetic as described above, wherein the anti-CMKLR1 antibody or antigen-binding fragment is combined with a second therapeutic agent.

[0355] The second therapeutic agent can be administered simultaneously or at different times with the anti-CMKLR1 compound. Depending on the nature of the second therapeutic agent, co-administration can produce a combination drug (product), also known as a "combo." A combo is a fixed-dose combination that contains two or more active pharmaceutical ingredients combined in a single dosage form, which is manufactured and distributed as a fixed dose. However, the dosage regimen and / or route of administration can also be different.

[0356] In a preferred embodiment, the second therapeutic agent is selected from the group consisting of: chemotherapeutic agents, radiotherapeutic agents, immunotherapeutic agents, cell therapy agents (such as CAR-T cells), antibiotics and probiotics.

[0357] In particular, the immunotherapeutic agents that can be used in the present invention are selected from: therapeutic vaccines (DNA vaccines, RNA vaccines or peptide vaccines), immune checkpoint blockers or activators, especially immune checkpoint blockers or activators of adaptive immune cells (T lymphocytes or B lymphocytes), or immunoconjugates (such as antibody-drug conjugates).

[0358] As used herein, the term "immunotherapeutic agent" specifically refers to agents that can divert cancer vaccines from the target biological phenomenon to effective therapeutic agents, including: T cell growth factors that increase the number and repertoire of naive T cells, growth factors that increase the number of dendritic cells (DCs), agonists that activate DCs and other antigen-presenting cells (APCs), adjuvants that allow and enhance cancer vaccines, agonists that activate and stimulate T cells, inhibitors of T cell checkpoint blockade, T cell growth factors that increase the growth and survival of immune T cells, and agents that inhibit, block or neutralize immunosuppressive cytokines derived from cancer cells and immune cells.

[0359] Many immune checkpoint blockers or activators are known in the art. In the present invention, examples of immune checkpoint blockers or activators of potentially useful adaptive immune cells (B or T lymphocytes) are: anti-PDL1, anti-PD1, anti-CTLA4, anti-SIRPa, anti-CD137, anti-CD2, anti-CD28, anti-CD40, anti-HVEM, anti-BTLA, anti-CD160, anti-TIGIT, anti-TIM-1 / 3, anti-LAG-3, anti-2B4, anti-OX40, anti-CD40 agonists, CD40-L, TLR agonists, anti-ICOS, ICOS-L and B cell receptor agonists, particularly anti-CD137 and anti-SIRPa. In a specific embodiment of the present invention, the second therapeutic agent is an anti-PDL1 or anti-PD1 compound, particularly an anti-PD1 compound, more particularly an anti-PD1 antibody. In a specific embodiment of the present invention, the second therapeutic agent is an anti-SIRPa compound, particularly an anti-SIRPa antibody.

[0360] The immunotherapeutic agent may also be an antibody targeting a tumor antigen, in particular selected from anti-Her2, anti-EGFR, anti-CD20, anti-CD19, and anti-CD52.

[0361] The antibody can be provided at an effective dose of about 1 ng / kg body weight to about 30 mg / kg body weight or higher. In specific embodiments, the dose can be 1 μg / kg to about 20 mg / kg, alternatively 10 μg / kg to 10 mg / kg, or 100 μg / kg to 5 mg / kg.

[0362] The term "effective dose" or "effective dosage" or "effective amount" is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The term "effective dose" is intended to include an amount sufficient to cure or at least partially prevent a disease and its complications or to alleviate the symptoms of the disease in a patient already suffering from the disease. The amount or dosage effective for this use will depend on the condition to be treated, the antibody construct delivered, the treatment situation and goal, the severity of the disease, previous treatments, the patient's clinical history and response to the therapeutic agent, the route of administration, the patient's size (weight, body surface or organ size) and / or condition (age and overall health) and the general state of the patient's own immune system. The appropriate dosage can be adjusted so that it can be administered to the patient once or multiple times to obtain the optimal therapeutic effect.

[0363] Doses for such purposes may be repeated as needed, for example daily, semiweekly, weekly, semimonthly, monthly, or as needed during recurring periods.

[0364] In another aspect, the present invention relates to a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof as described above and a pharmaceutically acceptable carrier.

[0365] As used herein, "pharmaceutical composition" is intended to include compositions suitable for administration to a subject or patient (e.g., a mammal, particularly a human). Typically, a "pharmaceutical composition" is sterile and generally does not contain contaminants that can cause adverse reactions in a subject (e.g., the compound in the pharmaceutical composition is pharmaceutical grade). A pharmaceutical composition can be designed to be administered to a subject or patient in need by many different routes of administration, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intrabronchial, and the like.

[0366] As used herein, "pharmaceutically acceptable carrier" is intended to include excipients, diluents, carriers, and adjuvants that can be used to prepare pharmaceutical compositions, which are generally safe, non-toxic, and not biologically or otherwise undesirable, and include excipients, diluents, carriers, and adjuvants that are acceptable for both veterinary and human pharmaceuticals. As used herein, "pharmaceutically acceptable carrier" includes more than one such excipient, diluent, carrier, and adjuvant.

[0367] In particular, the present invention relates to a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof as described above as an active ingredient and a pharmaceutically acceptable carrier.

[0368] In another aspect, the present invention relates to a therapeutic means (particularly a combination product means) comprising the following components as active ingredients: an anti-SIRPα antibody or antigen-binding fragment thereof or antigen-binding antibody mimetic as described above and a second therapeutic agent; wherein the active ingredients are formulated for separate, sequential or combined treatment, particularly for combined or sequential use.

[0369] In particular, the present invention relates to a combination product comprising an anti-CMKLR1 compound as described above and a second therapeutic agent, for simultaneous, separate or sequential use as a medicament.

[0370] In one embodiment, the invention relates to a combination product as described above, wherein the second therapeutic agent is selected from the group consisting of a chemotherapeutic agent, a radiotherapeutic agent, a cell therapy agent, an immunotherapeutic agent, an antibiotic and a probiotic.

[0371] In one embodiment, the present invention relates to a combination product as described above, wherein the immunotherapeutic agent is selected from: a therapeutic vaccine, an immune checkpoint blocker or activator, in particular an immune checkpoint blocker or activator of adaptive immune cells (T lymphocytes and B lymphocytes), and an antibody-drug conjugate.

[0372] In one embodiment, the present invention relates to a combination product as described above, wherein the immune checkpoint blocker or activator of the adaptive immune cells (T lymphocytes and B lymphocytes) is selected from: anti-PDL1, anti-PD1, anti-SIRPA, anti-CTLA4, anti-CD137, anti-CD2, anti-CD28, anti-CD40, anti-HVEM, anti-BTLA, anti-CD160, anti-TIGIT, anti-TIM-1 / 3, anti-LAG-3, anti-2B4 and anti-OX40, anti-CD40 agonists, CD40-L, TLR agonists, anti-ICOS, ICOS-L and B cell receptor agonists, in particular selected from: anti-PDL1, anti-PD1 and anti-CD137. In a specific embodiment of the present invention, the second therapeutic agent is an anti-PDL1 or anti-PD1 compound, in particular an anti-PD1 compound, more particularly an anti-PD1 antibody. In a specific embodiment of the present invention, the second therapeutic agent is an anti-SIRPa compound, in particular an anti-SIRPa antibody.

[0373] In one embodiment, the immunotherapeutic agent is an antibody targeting a tumor antigen, particularly selected from the group consisting of anti-Her2, anti-Her2, EGFR, anti-CD20, anti-CD19, and anti-CD52.

[0374] In one aspect, the invention relates to a combination as described above for simultaneous, separate or sequential use in the treatment of any condition susceptible to improvement or prevention by a change in macrophage polarization towards anti-inflammatory macrophages.

[0375] In one embodiment, the present invention relates to a method of treating any condition susceptible to improvement or prevention by a change in macrophage polarization to anti-inflammatory macrophages in a subject in need thereof, the method comprising simultaneously, separately or sequentially administering to the subject an effective amount of a combination product as described above.

[0376] In one embodiment, the invention relates to the use of a combination as described above for the preparation of a medicament for the treatment of any condition susceptible to improvement or prevention by a change in macrophage polarization to anti-inflammatory macrophages.

[0377] On the one hand, the present invention relates to a combination product as described above for simultaneous, separate or sequential use in the treatment of pathological types or for vaccination, said pathological types being selected from: inflammatory diseases, including but not limited to acute inflammatory diseases and chronic inflammatory diseases, inflammatory bowel disease, Crohn's disease, asthma, keratoconjunctivitis, periodontal disease, eczema, colitis, in particular ulcerative colitis or idiopathic colitis, diabetes (in particular type 1 diabetes), peritonitis, psoriasis, epithelial cancer (in particular breast or colon cancer), cancer, metastatic cancer, lung cancer, degenerative diseases, infectious diseases (in particular sepsis), autoimmune diseases.

[0378] In one embodiment, the present invention relates to a method for treating a pathological type in a subject in need thereof, wherein the pathological type is selected from: inflammatory diseases, including but not limited to acute inflammatory diseases and chronic inflammatory diseases, inflammatory bowel disease, Crohn's disease, asthma, keratoconjunctivitis, periodontal disease, eczema, colitis, especially ulcerative colitis or spontaneous colitis, diabetes (especially type 1 diabetes), peritonitis, psoriasis, epithelial cancer (especially breast epithelial cancer or colon epithelial cancer), cancer, metastatic cancer, lung cancer, degenerative diseases, infectious diseases (especially sepsis), autoimmune diseases, the method comprising administering to the subject simultaneously, separately or sequentially an effective amount of a combination product as described above.

[0379] In one embodiment, the present invention relates to the use of a combination product as described above for the preparation of a medicament for the treatment of a pathological type selected from: inflammatory diseases, including but not limited to acute inflammatory diseases and chronic inflammatory diseases, inflammatory bowel disease, Crohn's disease, asthma, keratoconjunctivitis, periodontal disease, eczema, colitis, in particular ulcerative colitis or idiopathic colitis, diabetes (in particular type 1 diabetes), peritonitis, psoriasis, epithelial cancer (in particular breast epithelial cancer or colon epithelial cancer), cancer, metastatic cancer, lung cancer, degenerative diseases, infectious diseases (in particular sepsis), autoimmune diseases.

[0380] The present invention also relates to a method for screening anti-CMKLR1 compounds, comprising the following steps:

[0381] a) providing a compound selected from an antibody or antigen-binding fragment thereof, a chimeric antibody, or a humanized antibody, the compound comprising:

[0382] An antibody heavy chain variable domain comprising three CDRs (i.e., VHCDR1, VHCDR2, and VHCDR3), wherein:

[0383] VHCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 4 or a mutant sequence thereof (in which amino acid residues are substituted); and

[0384] VHCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 6 or a mutant sequence thereof (wherein an amino acid residue is substituted); and

[0385] VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 144, or SEQ ID NO: 148, or a mutant sequence thereof (wherein the amino acid residues are substituted such that the amino acid residues at positions 1 and 2 of SEQ ID NO: 8 are L and I, respectively); and

[0386] An antibody light chain variable domain comprising three CDRs (i.e., VLCDR1, VLCDR2, and VLCDR3), wherein:

[0387] VLCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 12 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0388] VLCDR2 comprises or consists of the amino acid sequence shown in SEQ ID NO: 14 or a mutant sequence thereof (wherein the amino acid residues are substituted); and

[0389] VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or a mutant sequence thereof (wherein the amino acid residues are substituted);

[0390] b) testing the ability of the compound to act as an RvE1-like agonist of CMKLR1, in particular testing the ability of the compound to promote the resolution of inflammation, in particular against myeloid cells; optionally,

[0391] c) detecting the binding ability of the compound to an epitope located within the third loop E3 of CMKLR1, in particular the binding ability of the compound to a polypeptide comprising the amino acid residues of SEQ ID NO: 2 or SEQ ID NO: 152, by BLITZ assay or biosensor; alternatively,

[0392] d) detecting the ability of the compound to not compete for the binding of chemokine to CMKLR1;

[0393] e) when the binding capacity of the compound measured in step c) is at least 10E-8 KD:

[0394] i) after the compound binds to CMKLR1, detecting the ability of the compound to induce Akt phosphorylation and / or Erk phosphorylation, particularly the ability to induce Akt phosphorylation and Erk phosphorylation;

[0395] ii) testing the ability of the compound to enhance the secretion of anti-inflammatory cytokines (particularly IL-10, more particularly IL-10 and CCL17), and / or the ability of the compound to inhibit or reduce the secretion of pro-inflammatory cytokines (particularly IL-12), particularly on myeloid cells expressing CMKLR1; and / or

[0396] iii) testing the ability of the compound to promote macrophage polarization towards anti-inflammatory M2 macrophages, in particular the ability of the compound to enhance the secretion of the phenotypic marker CD200R; and / or

[0397] iv) testing the ability of the compound to inhibit dendritic cell activation and / or proliferation.

[0398] The ability of the compound can be tested according to the examples disclosed in the embodiments of the present disclosure.

[0399] The present invention also relates to polynucleotides encoding the anti-CMKLR1 compounds as described herein. To this end, the present invention also relates to nucleic acid molecules or groups of nucleic acid molecules, more particularly, isolated nucleic acid molecules and / or recombinant nucleic acid molecules, which encode any of the anti-CMKLR1 compounds disclosed herein, more particularly, which encode the heavy chain variable domain and the light chain variable domain:

[0400] the heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42; and / or

[0401] The light chain variable domain comprises or consists of the following amino acid sequence: SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60 or SEQ ID NO:61.

[0402] The nucleic acid molecule may further comprise regulatory sequences for transcription and expression of the encoded heavy chain variable domain and / or light chain variable domain, such as, but not limited to, enhancers, silencers, promoters, in particular expression promoters, and signal peptides.

[0403] The present invention also relates to vectors comprising polynucleotides disclosed herein or comprising nucleic acid molecules disclosed herein. As used herein, a vector is a nucleic acid molecule that is used as a vehicle for transferring genetic material into a cell. In a preferred embodiment, the vector allows the expression of the polynucleotides inserted into the vector. The term "vector" includes plasmids, viruses, cosmids, and artificial chromosomes. Typically, a vector comprises an origin of replication, a multiple cloning site, and a screening marker. Typically, the vector itself is a nucleotide sequence (typically a DNA sequence) comprising an insert (transgene) and a larger sequence serving as a vector "backbone." In addition to the transgenic insert and the backbone, modern vectors may also comprise other features: promoters, gene markers, antibiotic resistance, reporter genes, targeting sequences, and protein purification tags. Vectors referred to as expression vectors (expression constructs) are particularly useful for the expression of transgenes in target cells and typically have control sequences.

[0404] On the other hand, the present invention relates to cells, isolated cells, host cells, isolated host cells or cell lines comprising the above-mentioned carriers. As used herein, these terms related to cells are intended to include any individual cell or cell culture, which can be or have been the recipients (recipients) of the carriers, exogenous nucleic acid molecules and polynucleotides encoding the antibody constructs of the present invention; and / or the recipients of the antibody constructs themselves. Each material can be introduced into the cell by means of transformation, transfection, etc. These terms are also intended to include the offspring or potential offspring of individual cells. Suitable host cells include prokaryotic cells or eukaryotic cells, and also include but are not limited to bacteria, yeast cells, fungal cells, plant cells and animal cells, such as insect cells and mammalian cells, such as mouse cells, rat cells, rabbit cells, macaque cells or human cells.

[0405] The above figures and examples are presented to provide a complete disclosure and description of how to make and use the present invention to those of ordinary skill in the art, without any intention to limit the scope of the invention as the inventors believe, nor are they intended to represent that the following experiments are all or only experiments performed. Although the present invention has been described with reference to specific embodiments of the present invention, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the present invention. In addition, many modifications may be made to adapt specific circumstances, materials, compositions of matter, processes, process steps or steps to the purpose, spirit and scope of the present invention. All of these modifications are intended to fall within the scope of the appended claims.

[0406] Example

[0407] Production and screening of anti-CMKLR1 antibodies

[0408] Several antibodies were synthesized with different CDR sequences within the heavy and light chain variable domains. The different antibodies were tested for their ability to induce dendritic cell maturation and differentiation toward either pro-inflammatory or anti-inflammatory pathways. Given the differing results, the inventors compared the amino acid sequences of the resulting heavy and light chain variable domains and determined that amino acid differences within HCDR3 might be important for the experimental results. They selected antibody 2G1 (SEQ ID NO: 10 and SEQ ID NO: 17) to assess its ability to resolve inflammatory states, at least during the resolution phase.

[0409] like Figure 1 As shown, 2G1 and 1G1 antibody (another synthetic antibody) cells can inhibit the activation and / or maturation of DCs in a more potent manner than cells with other synthetic antibodies (3G1 and 4G1) against pro-inflammatory pathways, as the detection levels of DCs expressing CD103 and IAb treated with 2G1 were lower than those treated with C7 antibody (the method used in this assay is described in Example 10.2). Figure 1 As shown in E and 1F, cell viability was enhanced after treatment with 1G1 or 2G1 antibodies compared to cells treated with other antibodies, including C7 or resolvin E1.

[0410] 2G1 was humanized using a CDR splicing method in silico, which is a well-known humanization method in the art. The following table describes the obtained CDR and FR regions and the humanized sequences of the variable heavy chain and variable light chain.

[0411] Table 1: Wild-type sequences of the heavy chain variable domains of anti-CMKLR1 compounds (SEQ ID NO:4,SEQ ID NO: 6 and SEQ ID NO: 144) and humanized CDR (other SEQ ID).

[0412] <![CDATA[ SEQ ID NO ]]> <![CDATA[ sequence ]]> 4 VHCDR1 WT GFTFSSYGMS 6 VHCDR2 WT TINRYGGSTYYPDSVKG 144 VHCDR3 WT LIYYGNEGDS 62 VHCDR1 GFTFSSYAMS 63 VHCDR1 GYTFTSYGMS 64 VHCDR1 GYTFTSYAMS 65 VHCDR1 GYTFTSYAMN 66 VHCDR2 TINRYGGSTYYAASVKG 67 VHCDR2 TISRSGGSTYYAASVKG 68 VHCDR2 TINRYGGSTYYPDSFKG 69 VHCDR2 TINRYGGSTYYAQKFQG 70 VHCDR2 IINRNGGSTYYAQKFQG 71 VHCDR2 TINRYGGSPYYAQGFTG 72 VHCDR2 TINRYGGNPYYAQGFTG 145 VHCDR3 LIYYGNEGES 146 VHCDR3 LIYYGNEGDT 147 VHCDR3 LIYYGNEGET

[0413] Table 2: Wild-type sequences of light chain variable domains of anti-CMKLR1 compounds (SEQ ID NO: 12, SEQ ID NO: 14 and SEQ ID NO: 15) and humanized CDRs (SEQ ID NO: 76 to SEQ ID NO: 89)

[0414] SEQ ID No sequence 12 VLCDR1 WT SASSSVSFMH 14 VLCDR2 WT DTTKLTS 15 VLCDR3 WT QQWNSKPPLT 76 VLCDR1 RASQSVSFMH 77 VLCDR1 RASQSVSFLH 78 VLCDR1 RASQGISFLA 79 VLCDR1 RASQSVSFLA 80 VLCDR1 RASQSISFLH 81 VLCDR2 DATKLTS 82 VLCDR2 DASKLTS 83 VLCDR2 DASKLES 84 VLCDR2 DATKSTS 85 VLCDR2 DASKSFS 86 VLCDR2 DATKRTT 87 VLCDR2 DASKSTS 88 VLCDR2 DASKRTT 89 VLCDR3 QQWQSKPPLT

[0415] Table 3: Wild-type, humanized, and consensus FR sequences of the heavy chain variable domains of anti-CMKLR1 compounds

[0416] SEQ ID No sequence 90 HFR1 WT EVQLVASGGGLVQPGGSLKLSCAAS 91 HFR1 / 1 EVQLVESGGGLVQPGGSLRLSCAAS 92 HFR1 / 2 QVQLVQSGAEVKKPGASVKVSCKAS 93 HFR1 / 3 QVQLVQSGSELKKPGASVKVSCKAS 94 HFR2 WT WVRQTPDRRLELVA 95 HFR2 / 1 WVRQAPGKGLELVA 96 HFR2 / 2 WVRQAPGQGLELVA 97 HFR2 / 3 WVRQAPGQGLELVA 98 HFR2 / 4 WVRQAPGKGLELVS 99 HFR2 / 5 WVRQAPGQGLELVG 100 HFR2 / 6 WVRQAPGQGLELMG 101 HFR3 WT RFTISRDNAKNTLYLQMSSLKSEDTAMYYCPR 102 HFR3 / 1 RFTISRDNSKNTLYLQMNSLRAEDTAVYYCPK 103 HFR3 / 2 RVTITRDNSTSTLYMELSSLRSEDTAVYYCPR 104 HFR3 / 3 RFVISRDNSVSTLYLQISSLKAEDTAVYYCPR 105 HFR3 / 4 RVTITRDTSTSTVYMELSSLRSEDTAVYYCPR 106 HFR3 / 5 RFVISRDTSVSTAYLQISSLKAEDTAVYYCPR 107 HFR4 WT WGQGTTLTVSS 108 HFR4 WGQGTLVTVSS 19 HFR1 XVQLVXSGXGXXXXPGXSXXXSCXAS 125 HFR2 shares WVRXTPXXXLELVA 126 HFR3 shares RXXIXRDNXXXTLYXXXSSLXXEDTAXYYCPR 127 HFR4 shares WGQGTXXTVSS

[0417] Table 4: Wild-type, humanized, and consensus FR sequences of the light chain variable domains of anti-CMKLR1 compounds

[0418] SEQ ID No sequence 109 LFR1 WT / 1 QIVLTQSPAIMSASPGEKVTMTC 110 LFR1 / 1 AIQLTQSPSSLSASVGDRVTITC 111 LFR1 / 2 EIVLTQSPDFQSVTPKEKVTITC 112 LFR1 / 3 EIVLTQSPATLSLSPGERATLSC 113 LFR2 WT / 1 WYQQKSGTSPKRWIY 114 LFR2 / 1 WYQQKPGKAPKRWIY 115 LFR2 / 2 WYQQKPDQSPKRWIY 116 LFR2 / 3 WYQQKPGQAPRRWIY 117 LFR2 / 4 WYQQKPGKAPKRLIY 118 LFR3 WT / 1 GVPARFSGSGSGTFYSLTISSMEAEDAATYYC 119 LFR3 / 1 GVPSRFSGSGSGTDYTLTISSLQPEDFATYYC 120 LFR3 / 2 GVPSRFSGSGSGtDYTLTINSLEAEDAATYYC 121 LFR3 / 3 GIPARFSGSGSGTDYTLTISSLEPEDFAVYYC 122 LFR3 / 4 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC 132 LFR3 / 5 GVPSRFSGSGSGTDFTLTINSLEAEDAATYYC 123 LFR4 WT / 1 FGPGTKLELK 124 LFR4 / 1 FGGGTKVEIK 128 LFR1 total / 1 XIXLTQSPXXXSXXXXXTXTC 129 LFR2 total / 1 WYQQKXXXXPXRWIY 130 LFR3 total / 1 GXPXRFSGSGSGTXYXLTIXSXXXEDXAXYYC 131 LFR4 total / 1 FGXGTKXEXKR

[0419] Table 5: Heavy chain sequences of anti-CMKLR1 compounds

[0420]

[0421] Table 6: Light chain sequences of anti-CMKLR1 compounds

[0422]

[0423] Examples of the therapeutic efficacy of anti-CMKLR1 antibody therapy in preclinical models of autoimmune and inflammatory diseases

[0424] Example 1: DSS-induced colitis

[0425] Colitis was induced in 8- to 10-week-old C57BI / 6 male mice for 6 days by adding 2% (w / v) DSS to sterile drinking water ad libitum. Treatment was intraperitoneally injected with isotype control hIgG1 (10 μg / mouse), RvE1 (1 μg / mouse), or 2G1 antibody (10 μg / mouse) three times daily for 5 days. Colitis follow-up parameters, consisting of body weight and stool scores (0: normal stool; 4: bloody stool), were performed daily. At euthanasia, colon length, an indicator of pathology severity, was measured. The resolution index under different conditions was determined as described by Bannenberg et al. (2005).

[0426] result: Figure 2 The DSS animal model shown is an acute inflammation model. Figure 1 The animals treated with anti-CMKLR1 antibodies were in better overall condition than those receiving control antibodies or the resolvin RvE1. Figure 2 A) Significant reduction and stool score ( Figure 2B) was significantly better. Figure 2 C and 2D), animals receiving anti-CMKLR1 or RvE1 showed similar results.

[0427] Example 2: TNBS-induced colitis

[0428] On day 0, colitis was induced in 8- to 10-week-old C57BI / 6 male mice by intrarectal injection of 200 μL of 5% haptenized TNBS in 50% ethanol. Treatment was intraperitoneally injected with either RvE1 (1 μg / mouse) once daily or 2G1 antibody (10 μg / mouse) twice daily for 3 days. Colitis follow-up parameters, consisting of body weight and stool scores (0: normal stool; 4: bloody stool), were performed daily (data not shown). After euthanasia, colon length, an indicator of pathology severity, was measured.

[0429] Results: TNBS-induced colitis is another acute inflammatory model. Figure 3 The results showed that the colon lengths of animals treated with anti-CMKLR1 or RvE1 were the same as those of normal (wt) animals. However, the colon lengths of animals treated with the isotype control were shortened. These results confirm that anti-CMKLR1 antibodies, like RvE1, have therapeutic potential in this mouse model of acute inflammation.

[0430] Example 3: IL-10KO model - spontaneous colitis model

[0431] IL-10KO mice develop spontaneous colitis starting at 20 weeks of age, primarily due to a lack of regulatory T cell function (through IL-10 secretion in the intestine). IL-10KO mice were followed three times a week starting at 18 weeks of age to maintain weight loss and stool consistency, which are clinical hallmarks of this pathology. When weight loss exceeded 5% and stool scores were greater than or equal to 1, anti-CMKLR1 antibody (2G1) or isotype control (hIgG1) was injected intraperitoneally for 2 weeks (25 μg / injection, three times a week).

[0432] Results: The chronic inflammation model was used to study the efficacy of anti-CMKLR1 antibody treatment. Figure 4 Shown is an analysis of the percent body weight loss when animals were treated with isotype control or anti-CMKLR1 antibody ( Figure 4 A) and stool scores ( Figure 4 B) The results show that animals treated with anti-CMKLR1 antibodies lost less weight and had better stool scores than animals receiving isotype controls. Therefore, anti-CMKLR1 antibodies appear to have potential for treating chronic inflammatory diseases.

[0433] Example 4: Preclinical Model of Type 1 Diabetes: Mouse NOD Model

[0434] Eight-week-old NOD female mice were obtained from Charles River Laboratory. These mice developed spontaneous type 1 diabetes between 12 and 20 weeks. Diabetes induction can be measured by hyperglycemia. When blood glucose was between 180 mg / dL and 234 mg / dL, anti-CMKLR1 and isotype control were administered intraperitoneally at 20 μg / injection three times a week for two weeks. When blood glucose was above 600 mg / dL (corresponding to irreversible diabetes), mice were euthanized.

[0435] Results: This type I diabetes model is also considered a mouse autoimmune disease model. Figure 5 As shown in Figures AC, animals treated with anti-CMKLR1 antibodies showed improved survival and nearly normoglycemia as measured by blood glucose concentrations. This recovery appeared stable over time, suggesting that previously ill animals may fully recover. Anti-CMKLR1 antibodies restored glucose tolerance.

[0436] Example 5: Imiquimod-induced psoriasis-like skin inflammation

[0437] C57BI / 6 male mice (8 to 10 weeks old) were used Cream (known to induce psoriasis in mice). Mice received a topical dose of Aldara on the shaved back and left ear daily for 6 consecutive days. Treatment was intraperitoneally injected with RvE1 (1 μg / mouse) and PBS daily. The ears were measured daily ( Figure 6 A) and back ( Figure 6 B) thickness until it recovers.

[0438] Results: Ear and skin thickness were reduced in animals treated with the resolvin RvE1 compared to animals treated with the control molecule. These results suggest that agonist anti-CMKLR1 antibody therapy has potential application in mouse models of psoriasis, a representative autoimmune disease.

[0439] Example 6: Therapeutic efficacy of anti-CMKLR1 antibodies in a preclinical model of sepsis (mouse peritonitis model)

[0440] Conventional peritonitis was induced (1 mg (1 mL) / mouse). Anti-CMKLR1 prophylactic injections, including RvE1 (1 μg / mouse) and 2G1 antibodies (10 μg / mouse), were administered 5 minutes prior to Zymosan A injection. Peritoneal polymorphonuclear neutrophils (PMNs) and macrophages were collected 2, 4, 8, 16, 24, and 48 hours after Zymosan A injection, counted by flow cytometry, and the resolution index was determined (Bannenberg et al., 2005).

[0441] result: Figure 7 Results showing PMN (7A) and macrophage (7B) numbers and resolution index (7C) indicate that animals treated with RvE1 or anti-CMKLR1 antibodies exhibited similar results, with slightly fewer PMN and macrophage numbers and higher resolution indexes compared to isotype controls. In sepsis, even slight differences can be therapeutically important. Therefore, these results strongly support the potential use of anti-CMKLR1 antibodies in sepsis.

[0442] Example 7: Therapeutic efficacy of anti-CMKLR1 antibody therapy in preclinical models of cancer

[0443] Example 7.1 : Effects of anti-CMKLR1 antibody on primary tumor growth and the development of lung metastases in an orthotopic breast epithelial carcinoma model.

[0444] Mice were anesthetized with 3% isoflurane. The abdomen of the mice was shaved and 4T1 cells (250,000) in 50mL PBS were injected into the mammary glands using an insulin syringe (30G). On the 4th and 7th days, anti-CMKLR1 antibody (2G1) or anti-41BB antibody (3H3) or both antibodies (10 μg / injection) were injected twice; a control antibody (100 μg / injection) in PBS was injected intraperitoneally three times a week (for a total of three weeks). In a second study measuring lung metastasis after the development of mammary epithelial carcinoma, animals were treated with 0.8 mg / kg anti-CMKLR1 antibody or control antibody (100 μg / injection) three times a week for three weeks.

[0445] Results: As Figure 8 As shown in A, there was no improvement in tumor growth in animals treated with a single compound (2G1 or 3H3) compared to animals receiving an isotype control antibody. However, animals treated with a combination of anti-CMKLR1 and anti-41BB antibodies showed a significant reduction in tumor growth in the mammary epithelial cancer model (p < 0.01). Since this mammary epithelial cancer model is a severely aggressive model, the results are considered positive. Figure 8As shown in Figure B, the effect of the anti-CMKLR1 compound on lung metastasis was shown by bioluminescent imaging, and it can be seen that anti-CMKLR1 treatment reduced lung metastasis compared to animals treated with a control antibody. Analysis of lymph node metastasis showed that there was no metastasis in animals treated with the anti-CMKLR1 compound, while two animals in the control group had metastasis (data not shown). These results indicate that anti-CMLKR1 antibodies with agonist activity that mimics RvE1 have an anti-metastatic effect. In this model, the antibodies of the present invention did not show any significant effect on the development of primary tumors. However, the results showed that improved results were obtained when animals were treated with a combination of anti-CMKLR1 and anti-41BB antibodies.

[0446] Example 7.2 : Therapeutic effects on tumor growth in a colon cancer model

[0447] Eight-week-old C57bl / 6J males were anesthetized with 3% isoflurane. The abdomen of the mice was shaved and MC38 cells (0.5×10 6 Another model was used in which 8-week-old Balb / c male mice were anesthetized with 3% isoflurane. The abdomen of the mice was shaved and CT26 cells (1×10 CT26 cells / mouse) in 50 mL PBS were injected subcutaneously with an insulin syringe (30G). 6 cells / mouse).

[0448] Starting from day 4 after tumor inoculation, agonist anti-CMKLR1 antibody (2G1) or anti-SIRPa antibody (p84-anti-mouse SIRPa from Merck Millipore) (SIRPa is a novel checkpoint inhibitor) was injected intraperitoneally once a week (20 μg / injection) alone or in combination for 3 weeks.

[0449] Results: As Figure 9 As shown in Figures AC, in the CT26 cancer model, anti-CMKLR1 antibody alone showed no clinical effect on tumor progression compared with the control group ( Figure 9 B), anti-SIRPα alone also showed no clinical effect on tumor progression ( Figure 9 A). However, surprisingly, the combination of the two compounds was able to inhibit tumor growth in a timely manner ( Figure 9 C) In Figure 9 In another mouse model of colon epithelial cancer, shown in D, anti-CMKLR1 indeed demonstrated efficacy in inhibiting tumor growth compared to an isotype control. Taken together, these results from two different models of colon epithelial cancer suggest that agonistic anti-CMKLR1 antibodies can be used alone or in combination with other therapeutic agents to prevent tumor development.

[0450] Example 8: Meta-analysis of CMKLR1 expression in biopsies of human patients with UC or CD treated with anti-TNFα or anti-α4β7 antibodies

[0451] In humans, the signaling networks underlying long-standing chronic gastrointestinal inflammation in chronic Crohn's disease (CD) and ulcerative colitis (UC), the two major forms of inflammatory bowel disease (IBD), are poorly understood. Based on an analysis of nearly 500 IBD patients and 100 controls, the inventors here report that CMKLR1 transcripts accumulate in inflamed colonic tissue of patients with severe IBD who are unresponsive to immunosuppression / corticosteroids and immunotherapies such as anti-TNFα (infliximab) or anti-α4β7 integrin (vedolizumab).

[0452] The inventors first analyzed mucosal CMKLR1 transcript expression by meta-analyzing publicly available transcriptome datasets from three cohorts of UC patients (GSE16879 (Arijs et al., 2009a) and GSE12251 (Arijs et al., 2009b), as well as GSE73661); colonic mucosal biopsies were obtained before (within one week) anti-TNF treatment in patients refractory to corticosteroids and / or immunosuppression. In these three cohorts, anti-TNF response was defined as histological healing 4 to 6 weeks after the first anti-TNF infusion (total: non-IBD controls n = 18, UC non-responders n = 41, and UC responders n = 28).

[0453] Results: The analysis showed that CMKLR1 transcript expression was significantly increased in colonic biopsies from primary UC non-responders before and after anti-TNF treatment compared with non-IBD controls or UC patients before anti-TNF treatment (who would respond to anti-TNF treatment). Figure 10 A). In colonic or ileal biopsies from patients with Crohn's disease, mucosal CMKLR1 expression was also significantly increased before and after anti-TNF treatment in patients who were non-responsive to anti-TNF compared with non-IBD controls or future responders (non-IBD controls n = 24, CD non-responders n = 17, and CD responders n = 20; GSE16879 (Arijs et al., 2009a)) ( Figure 10 Finally, gene expression analysis of the colonic mucosa of a cohort of UC patients treated with anti-α4β7 (vedolizumab) (GSE7366146) also confirmed that CMKLR1 expression was significantly increased in non-responders before and after vedolizumab treatment ( Figure 11 ).

[0454] In summary, the meta-analysis showed that CMKLR1 is overexpressed in inflamed tissues of IBD patients, particularly in patients who do not respond to current immunosuppression or immunotherapy (even before starting treatment). Our meta-analysis provides evidence that CMKLR1 expression in the colon (ileum for CD patients) of patients with refractory UC or CD may conversely predispose these patients to treatment with agonistic anti-CMKLR1 antibodies (e.g., the antibodies of the present invention).

[0455] Example 9: CMKLR1 expression and antibody binding studies

[0456] ELISA binding CMKLR1( Figure 12 )

[0457] CMKLR1 peptide (273NH2-PYHTLNLLELHHTAMPGSVFSLGLPLATALAIA-COOFI305) (SEQ ID NO: 18) (5 μg / ml) was coated overnight in borate buffer. Saturation was performed with PBS-Tween 0.1%-Gelatin 0.25% at 37°C for 2 hours. 2G1 or hIgG1 antibodies were then added at various concentrations for 2 hours at 37°C. Peroxidase-conjugated secondary antibodies (0.8 μg / ml) were then added at 37°C for 1 hour, and color was developed using TMB substrate. The colorimetric reaction was read using TECAN.

[0458] FACS determination of CMKLR1 expression ( Figure 13 A)

[0459] The cells were resuspended in PBS-FBS-EDTA and incubated on ice with Fc blocker (1 / 50) for 30 minutes. Monocytes, macrophages, and dendritic cells were stained with A488-labeled 2G1 (5 μg) or A488-labeled hIgG1 (5 μg).

[0460] Western Blot Analysis of CMKLR1 ( Figure 13 B)

[0461] After protein migration and transfer as described above, 2G1 antibody (10 μg / membrane) was incubated overnight at 4°C and developed with a peroxidase-conjugated secondary antibody (1:2000). CMKLR1 expression was then detected using chemiluminescence and an image reader. Western blot images were quantified using Multi Gauge software.

[0462] result: Figure 12The results shown demonstrate that the anti-CMKLR1 antibody clone 2G1 binds to the peptide that forms loop EL3 of CMKLR1. CMKLR1 expression in different cell lines was assessed by FACS and Western Blot using the 2G1 antibody. The results showed that human tumor T cell lines Trp1 and U937, as well as transduced CMKLR1 CHO cells, human lung fibroblast cell line MRC5, and human NK cell line NKL expressed CMKLR1 ( Figure 13 ).

[0463] Example 10: Study of CMKLR1 expression in myeloid lineage

[0464] Example 10.1 :Differentiation and polarization of human monocytes

[0465] Monocytes were collected from PBMCs of buffy coats of healthy volunteers and isolated by magnetic separation or elutriation. Monocytes were then cultured with different cytokine mixtures to produce differentiated non-polarized macrophages or polarized macrophages. This protocol made it possible to produce polarized differentiated macrophages to have either pro-inflammatory macrophages (M1) or anti-inflammatory macrophages (M2) in different wells. Monocytes were plated at 0.5×10 6 Cells were plated at a concentration of 100 μL / mL in complete RPMI (RPMI containing 10% FBS, 1% glutamine, and 1% antibiotics), and 500 μL of the cell suspension was plated in each well of a 24-well plate. 100 ng / mL M-CSF was added to the culture medium for differentiation of the cells. The cells were incubated for 5 days, and the culture medium was replaced with fresh culture medium supplemented with 100 ng / mL M-CSF on the 3rd day. For the polarization phase, LPS-IFNg solution (100 ng / mL LPS and 20 ng / mL IFNg) supplemented with isotype control (rnlgd or hIgG4) (2 μg / ml) or anti-CMKLR1 antibody (2 μg / ml) (2G1 or 2G4, H6, BZ332, or 84939) or supplemented with C15 peptide (10 nM) or RvE1 (10 ng / ml) was added over a 3-day period to generate M1 macrophages. M1-IFNg macrophages can also be generated by adding only IFNg (20 ng / mL) to the culture medium. For M2 polarization, cells are incubated with 20 ng / mL IL-4. Following differentiation and / or polarization, phenotype and functional cytokine / chemokine release are investigated by FACS analysis, ELISA, and Western Blot.

[0466] Example 10.2 :Isolation and differentiation of mouse macrophages and DCs

[0467] -Isolation of mouse bone marrow-derived macrophages

[0468] Bone marrow cells were harvested and cultured for 5 days in RPMI medium supplemented with 10% FBS, glutamine, and antibiotics and containing 100 ng / mL macrophage colony stimulating factor (M-CSF) to induce macrophage differentiation. Macrophages were harvested and cultured for 2 days with IFNg (20 ng / ml) and LPS (100 ng / ml) to induce M1 polarization or IL-4 (20 ng / ml) to induce M2 polarization. During macrophage polarization, therapeutic agents were added at 2 μg / ml.

[0469] -Generate bone marrow-derived dendritic cells

[0470] Bone marrow cells were collected and cultured in RPMI medium supplemented with 10% FBS, glutamine, and antibiotics. Dendritic cell differentiation was induced with 20 ng / ml GM-CSF for 7 days. Immature dendritic cells (iDCs) were then collected and cultured with LPS (100 ng / ml) for 24 hours to induce maturation from iDCs to mDCs. During the differentiation and maturation period, therapeutic agents were added at 2 μg / ml.

[0471] After mouse M1 or M2 differentiation as described above, cells were incubated in the presence of culture medium, isotype control, anti-CMKLR1 antibody: clone H6 and BZ194, C15 peptide, 2G1 (target anti-CMKLR1 antibody) or RvE1 were used. The secretion of IL-10, CCL17 and IL-12p40 was then assessed by ELISA. Cytokine secretion in the supernatant was measured using an ELISA kit purchased from BD. The supernatant was diluted 1 / 10 for IL-10 cytokine, 1 / 50 for CCL17 cytokine, and 1 / 100 for IL-12p40 cytokine.

[0472] -ELISA cytokine secretion study

[0473] Cytokine secretion was detected by ELISA according to the BD manufacturer's instructions. Briefly, the supernatant was diluted with the appropriate buffer, coated overnight, and incubated with the capture antibody for 2 hours to saturate. Cytokine expression was then visualized using a biotin-conjugated detection antibody, and the signal was amplified using a biotin-streptavidin-conjugated peroxidase system. TMB provided by BD Bioscience was used as a substrate, and the colorimetric reaction was read using TECAN.

[0474] - Analysis of activated cell markers by FACS

[0475] Dendritic cells were resuspended in PBS-FBS-EDTA and stained with live cell dye and dead cell dye ( The cells were incubated with Fixable Dead Cell Stains Yellow (Life Technologies) for 30 minutes. The cells were then stained with CD11c-BV711, CD11b-APCCy7, 1 / Ab-APC, CD103-PerCPCy5.5, CCR7-V450, CD40-PeCy7, CD80-PE, and CD86-FITC (all provided by BD Pharmingen).

[0476] - Western Blot analysis of ERK / Akt

[0477] Mouse proinflammatory macrophages (M1) were generated from the bone marrow using M-CSF and polarized with IFN-γ (IFNg) and LPS. Briefly, bone marrow cells were collected by flushing the femur, cultured for 5 days with 100 ng / mL mM-CSF, and then polarized for 24 hours with 20 ng / mL IFNg and 100 ng / mL LPS. Then, FBS was removed with RPMI FBS2% culture medium for 24 hours. Finally, mouse M1 was treated with 2 μg / mL of 2G1 antibody for different times (5 minutes, 10 minutes, and 30 minutes). Cells were collected in RIPA buffer. Protein concentration was measured by BCA protein assay. Protein was denatured by heating at 95°C for 5 minutes and diluted with DTT and Laemmli solution. After migration and transfer, nitrocellulose membranes were blocked with 5% BSA in TBS-T for 2 hours. Anti-phospho-ERK and anti-phospho-Akt antibodies (1:1000) were incubated with the membrane overnight at 4°C and developed with peroxidase-conjugated secondary antibodies (1:2000). Western blot images were quantified using Multi Gauge software.

[0478] Example 10.3 :CMLKR1 expression in human blood monocytes and mouse bone marrow myeloid cells and neutrophils after inflammatory stimulation

[0479] Human monocytes were collected from PBMCs in buffy coats of healthy volunteers and isolated by magnetic separation or elutriation. Monocytes (CD14-positive cells) were then cultured in culture medium and treated with various proinflammatory stimuli for 16 or 48 hours: LPS (100 ng / ml), TNFa (10.000 U / ml), or IL-6 (20 ng / ml).

[0480] Mouse monocytes (CD11b+Ly6G-SSClow) and neutrophils (CD11b+Ly6G-SSClow) were harvested from bone marrow cells and cultured in RPMI medium supplemented with 10% FBS, glutamine, and antibiotics. The cells were then cultured with the medium and treated with different proinflammatory stimuli for 16 or 48 hours: LPS (100 ng / ml), TNFa (10.000 U / ml), or IL6 (20 ng / ml).

[0481] The expression of CMLKR1 was measured by FACS using commercially available anti-CMKLR1 antibodies (human anti-ChemR23: clone 84939 and mouse anti-ChemR23: clone 477806).

[0482] Result: Yes Figure 14 Analysis of CMKLR1 expression in the myeloid lineage of mice shown indicates that monocytes and type 1 and 2 macrophages as well as dendritic cells express the protein well. Figure 19 CMKLR1 expression in human monocytes and mouse bone marrow myeloid cells and neutrophils is shown. Inflammatory stimuli (such as LPS, TNFa or IL6) significantly increase expression (at least two-fold compared to control after 48 hours), confirming that CMKLR1 expression in myeloid cell lineages and overexpression during inflammation may represent a therapeutic approach to downregulate and / or induce resolution of inflammation. Figure 15 The study of inflammatory cytokines secreted by mouse M1 or M2 macrophages showed that only 2G1 clone (anti-CMKLR1 antibody) induced the production of anti-inflammatory cytokines IL-10 and CCL17 ( Figure 15 A and 15B), and reduced pro-inflammatory cytokines in M1 macrophages ( Figure 15 C). The results were different for cells treated with RvE1 or the C15 peptide at the C-terminus of chemokine (Cash et al., 2008), or even with other commercially available anti-CMKLR1 antibodies, suggesting a very interesting profile of the 2G1 antibody as an anti-inflammatory compound. The inventors then investigated cytokine expression in human myeloid cells obtained from donors and incubated with the different compounds. Figure 16 The results shown here indicate that 2G4 induces M2 polarization of cells compared to the control group and the commercially available CMKLR1 antibody C7, confirming the results obtained in mouse cells. Only the 2G1 antibody can induce the production of anti-inflammatory cytokines IL-10 and CCL17 ( Figure 16 A and 16B), and can reduce the secretion of IL-12p40 cytokine by M1 macrophages ( Figure 16 C). Finally, DC activation markers were analyzed by FACS. Figure 17The results shown here show that when cells were treated with RvE1 lipid or 2G1 antibody, the expression of CD80, CD86, CD103, CD40, and IAb was significantly reduced compared to vehicle or isotype. These results suggest that, like RVE1, 2G1 antibody can activate the CMKLR1 pathway in DCs. The inventors then analyzed the CMKLR1 activation pathway in mouse macrophages by Western blot. Figure 18 The anti-CMKLR1 antibody 2G1 was shown to induce activation of Akt and Erk proteins after 10 to 30 minutes of incubation. These results suggest that, like RvE1, the 2G1 antibody exhibits agonist properties at the CMKLR1 receptor.

[0483] Example 11: Competition studies of chemokine-induced CMKLR1 activation using anti-CMKLR1 antibodies

[0484] method

[0485] Chemokine-dependent B-inhibitors through the CMKLR1 receptor were measured by competition assay in the presence of anti-CMKLR1 antibodies. Protein recruitment:

[0486] The day before the assay, CHO-K1 CMKLR1 cells (Discover'X reference number 93-0313E2) were added to pre-warmed cell reagent and then plated in 96-well plates (Discover'X reference number 15-103) at 100 μl / well and incubated at 37°C in a humidified incubator with 5% CO2 for 48 hours. Anti-CMLKR1 antibody was diluted (22X, 1 μM to 1 nM, 7-point series of 3-fold dilutions) and cells were incubated with antibody for 30 minutes at 37°C. Cells were then stimulated with chemokine (2 nM or 6 nM) for 90 minutes at 37°C according to the supplier's protocol (Discover'X reference number 92-1036). After adding the working detection solution to the cells, luminescence was measured at 0.5 second intervals using a microplate reader.

[0487] Measuring Competition of Anti-CMKLR1 Antibodies with Chemokines in AMPc Production via the CMKLR1 Receptor:

[0488] On the day before the experiment, CHO-K1 CMKLR1 Gi cells (Discover'X reference number 95-0080C2) were added to the pre-warmed cell reagent, and then the cells (Discover'X reference number 15-103) were plated in 96-well plates at 100 μl / well and incubated in a 37°C, 5% CO2 humidified incubator for 24 hours.

[0489] At 37°C, chemokine agonists (6X, 10 -7 μM to 10 -10A mixture of chemokine (2 nM, 7-point series of 3-fold dilutions) (purchased from R&D Systems' Discover'X reference number 92-1036 or 2324-CM-025) and forskolin (40 μM) (cAMP activator) (Discover'X reference number 92-0005) was added to the cells; alternatively, the cells were preincubated with anti-CMKLR1 antibody for 30 minutes at 37°C (serial dilution: 6X, 1 μM to 1 nM, 7-point series of 3-fold dilutions). Then, a mixture of chemokine (2 nM) + forskolin (60 nM) was added to the cells at 37°C for 30 minutes. For cAMP detection, antibody reagent and cAMP working detection solution were added to the plate for 1 hour at room temperature, and then cAMP solution A was added and the cells were incubated in the dark for 3 hours at room temperature. Luminescence was measured using a microplate reader at 0.5 second intervals.

[0490] Results: To test whether the antibodies of the present invention are antagonists of chemokine-induced CMKLR1 activation, two assays were performed. Figure 20 Inhibition of chemokine-induced forskolin-dependent cAMP production Figure 20 As shown in A (black circles or white squares); compared with the control (grey diamonds), the anti-CMLKR1 antibodies of the present invention cannot restore such inhibition (black circles or white squares). Figure 20 The chemokine-induced β-arrestin activation shown in B shows that the anti-CMKLR1 antibodies of the present invention do not significantly alter chemokine-dependent β-arrestin activation (white circles, compared to black diamonds). The antibodies of the present invention do not have antagonist activity of the CMLKR1-chemokine interaction. In addition, the antibodies of the present invention cannot induce the chemokine-induced CMKLR1 signaling pathway, confirming that these antibodies are not agonists of chemokines in the CMLKR1 pathway.

[0491] Example 12: CD45Rb 高 T cell transfer chronic colitis mouse model

[0492] Methods: CD45Rb was isolated from the spleen of naive mice. 高 CD4 T cells were negatively selected by magnetic sorting and then sorted by ARIA FACS, and 0.5 × 10 6 Cells (in 100 μL PBS) were injected intraperitoneally into 6-week-old female Rag1 knockout mice. 高Starting on day 32 after CD4 T cell transfer, anti-CMKLR1 antibody (2G1) or isotype control was administered at 1 mg / kg three times a week for three weeks. Body weight was assessed three times a week to determine the change from baseline. *p < 0.05, *p < 0.01.

[0493] result: Figure 21 The percentage weight change of animals treated with anti-CMLKR1 antibodies or isotype controls over time after antibody administration is shown. Both groups showed the same initial weight change over the first 30 days after treatment, beginning to diverge at day 35. Mice treated with anti-CMKLR1 continued to gain weight, while control mice, in contrast, began to lose weight (indicating that the control group developed chronic colitis as predicted). The inventors used a third model of colitis (here, a chronic inflammatory model) to demonstrate that the anti-CMKLR1 antibodies of the present invention may be useful for treating chronic inflammatory and autoimmune diseases, such as colitis.

[0494] Example 13: Effect of Antitumor Effect on Overall Survival Rate of Mouse Liver Cancer Tumor Model

[0495] Methods: Mice were anesthetized with a xylazine / ketamine mixture. After laparotomy, tumor Hepa 1.6 cells (2.5×10 6 cells / 100 μL). Treatment was initiated 4 days after tumor injection. Anti-CMKLR1 antibody (2G1 clone) and hIgG1 isotype control were injected at 0.8 mg / kg three times per week for 2 weeks. Anti-PD1 monoclonal antibody (8 mg / kg) in PBS was injected intraperitoneally twice per week for 2 weeks. Combinations of anti-CMKLR1 and anti-PD1 antibodies (0.8 mg / kg and 8 mg / kg, respectively) were also tested. Overall survival was followed for 60 days, and the survival rate in each case is reported ( Figure 22 ).

[0496] Results: As Figure 22 As shown, only one of seven treated animals (15% of treated animals) treated with anti-CMKLR1 or anti-PD1 antibodies had prolonged survival, indicating a partial response (PR). However, animals treated with a combination of anti-PD1 and anti-CMKLR1 antibodies had a significantly improved survival rate (from 15% to 45%) (animals surviving 60 days after treatment), indicating a complete response (CR). This result suggests that the treatment combination (anti-PD1 / anti-CMKLR1 antibodies) has unexpected efficacy in HCC tumor models.

[0497] References:

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Claims

1. An anti-CMKLR1 compound, wherein the anti-CMKLR1 compound is selected from an antibody or antigen-binding fragment thereof, a chimeric antibody, a humanized antibody, or a modified antibody that specifically binds to CMKLR1, wherein: The anti-CMKLR1 compound comprises: An antibody heavy chain variable domain comprising three CDRs, namely VHCDR1, VHCDR2 and VHCDR3, wherein: VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64 or SEQ ID NO: 65; and VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71 or SEQ ID NO: 72; and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 8, SEQ ID NO: 73, SEQ ID NO: 74 or SEQ ID NO: 75; and An antibody light chain variable domain comprising three CDRs, VLCDR1, VLCDR2 and VLCDR3, wherein: VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79 or SEQ ID NO: 80; and VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87 or SEQ ID NO: 88; and VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO:

89.

2. The anti-CMKLR1 compound according to claim 1, wherein The anti-CMKLR1 compound is a resolvin E1-like agonist of CMKLR1; in particular, the anti-CMKLR1 compound is a pro-resolvation factor, in particular for myeloid cell lineage; in particular, the anti-CMKLR1 compound is an anti-human CMKLR1 compound.

3. The anti-CMKLR1 compound according to claim 1 or 2, wherein The anti-CMKLR1 compound specifically binds to an epitope located within the third extracellular loop (EL3) of CMKLR1; in particular, the anti-CMKLR1 compound specifically binds to a polypeptide comprising the amino acid residues of SEQ ID NO:

2.

4. The anti-CMKLR1 compound according to any one of claims 1 to 3, wherein The VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO:

148.

5. The anti-CMKLR1 compound according to any one of claims 1 to 4, wherein: VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 62 or SEQ ID NO: 63; and VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 70 or SEQ ID NO: 72; and VHCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 144 or SEQ ID NO:

148.

6. The anti-CMKLR1 compound according to any one of claims 1 to 5, wherein: VLCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO: 77; and VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 81 or SEQ ID NO: 84; and VLCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO:

89.

7. The anti-CMKLR1 compound according to any one of claims 1 to 6, wherein: the heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, or SEQ ID NO: 42; The light chain variable domain comprises or consists of the amino acid sequence shown in SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60 or SEQ ID NO:

61.

8. The anti-CMKLR1 compound according to any one of claims 1 to 7, wherein The anti-CMKLR1 compounds enhance the secretion of anti-inflammatory cytokines, in particular IL-10 and / or CCL17, more particularly IL-10, in vitro and / or in vivo, and / or the anti-CMKLR1 compounds inhibit or reduce the secretion of pro-inflammatory cytokines, in particular IL-12, in vitro and / or in vivo, particularly on myeloid cells expressing CMKLR1.

9. The anti-CMKLR1 compound according to any one of claims 1 to 8, wherein The anti-CMKLR1 compounds promote macrophage polarization toward anti-inflammatory M2 macrophages.

10. The anti-CMKLR1 compound according to any one of claims 1 to 9, wherein The anti-CMKLR1 compounds inhibit the activation and / or proliferation of dendritic cells.

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