Novel use of antibody specifically binding to LRR domain

Abstract

The LRR domain is a protein domain that functions as a structural platform mediating selective interactions with various molecules on the basis of repetitive leucine-rich sequences and a unique three-dimensional structure, and plays an important role in physiological and pathological processes, and thus can be used as a novel target for therapeutic agents for various metabolic diseases, neurological diseases, and fibrosis. Therefore, an antibody or antigen-binding fragment specifically binding to the LRR domain provided in the present invention can prevent or treat various metabolic diseases, neurological diseases, and fibrosis.

Description

New uses of antibodies that specifically bind to the LRR domain

[0001] The present invention relates to new uses of antibodies that specifically bind to the LRR (Leucine-Rich Repeat) domain, specifically for the treatment of metabolic diseases, neurological diseases and fibrosis.

[0002] Fibrosis occurs when the equilibrium between osteoclasts, which absorb bone, and osteoblasts, which form bone, is disrupted within the body. Osteoporosis, a representative form of fibrosis, arises from an imbalance in the regulation of bone homeostasis by osteoclasts and osteoblasts; as the activity of osteoclasts increases relative to that of osteoblasts, total bone mass decreases, causing bones to fracture easily even from minor impacts.

[0003] In addition, metabolic diseases are a collective term for diseases caused by metabolic disorders within the body, resulting from imbalances in carbohydrates, lipids, proteins, vitamins, minerals, and water. Among these, metabolic diseases caused by lipid imbalance include insulin resistance, fatty liver, diabetes mellitus, type 2 diabetes, hyperlipidemia, hypertriglyceridemia, dyslipidemia, cardiovascular disease, stroke, myocardial infarction, hyperglycemia, hyperinsulinemia, arteriosclerosis, hypertension, heart disease, obesity, liver disease, kidney injury, and hypertension.

[0004] Hyperlipidemia refers to a condition in which excessive fatty substances exist in the blood, accumulate on the blood vessel walls, cause inflammation, and consequently lead to cardiovascular disease; recently, abnormal blood lipid levels are also defined as dyslipidemia. While hyperlipidemia often occurs due to genetic factors leading to an increase in specific blood lipids, it can also be caused by other factors such as obesity, alcohol consumption, and diabetes.

[0005] Arteriosclerosis is a disease in which cholesterol is deposited on the innermost layer (endothelium) of blood vessels and endothelial cells proliferate, causing the blood vessels to narrow or become blocked, thereby disrupting blood flow; however, the exact cause of its occurrence has not been clearly identified.

[0006] One objective of the present invention is to provide novel uses for antibodies that specifically bind to the LRR domain, specifically for the prevention and treatment of obesity, neurological diseases, and fibrosis.

[0007] Another objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity, neurological diseases, and fibrosis, or a method for prevention and treatment comprising antibodies that specifically bind to the LRR domain as active ingredients.

[0008] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below.

[0009] In the present invention, the term “Lrig-1 protein” refers to a transmembrane protein composed of 1,091 amino acids present on the surface of immune cells, particularly regulatory T cells, and consists of a leucine-rich repeat (LRR) sequence located extracellularly or in the lumen, three immunoglobulin-like domains, a transmembrane sequence, and a cytoplasmic tail. The LRIG gene family includes LRIG1, LRIG2, and LRIG3, and the amino acids among them are highly conserved. The LRIG1 gene is highly expressed in normal skin and is expressed in basal and hair follicle cells, capable of regulating the proliferation of epithelial stem cells. Therefore, it plays an important role in maintaining epidermal homeostasis, and its absence can lead to the development of psoriasis or skin cancer. It has been reported that there is a high likelihood of developing into cancer cells when the chromosome 3p14.3 region where LRIG1 is located is cleaved, and indeed, it has been confirmed that the expression of LRIG1 is significantly reduced in renal cell carcinoma and cutaneous squamous cell carcinoma. Currently, LRIG1 is being suggested as a potential cancer inhibitor by degrading the protein through ubiquitination of EGFR (Epidermal growth factor receptor) via c-Cbl, thereby blocking signal transduction caused by the phosphorylation of MAPK and AKT located downstream of EGFR and involved in cell proliferation, and by inducing apoptosis by increasing the secretion of Caspase-8. For the purposes of the present invention, the Lrig-1 protein may be a protein present in humans or mice, but is not limited thereto.

[0010] In the present invention, the fragment of the LRIG protein may be composed of a sequence of amino acids, for example, 2 to 50; 6 to 45; or 10 to 44, of the amino acid sequence constituting the LRIG protein, preferably LRIG1, LRIG2, or LRIG3 protein, but is not limited thereto.

[0011] In one example of the present invention, the fragment of the LRIG protein may be a leucine-rich repeats protein (LRR) or a fragment thereof of the LRIG protein, preferably LRIG1, LRIG2, or LRIG3 protein, but is not limited thereto.

[0012] In another example of the present invention, the fragment of the LRIG protein may be an extracellular domain of the LRIG protein, preferably LRIG1, LRIG2, or LRIG3 protein, or a fragment thereof, but is not limited thereto.

[0013] In the present invention, the term “LRR domain (leucine-rich repeat domain)” refers to a protein structural motif characterized by repeat sequences rich in leucine residues, which is widely found in various biological species and is a functional domain primarily involved in protein-protein interactions. The LRR domain generally has a structure in which repeat units consisting of about 20 to 30 amino acids are arranged in series, and each repeat unit contains a conserved sequence motif. A representative consensus sequence is in the form LxxLxLxxNxL or a similar form, where “L” represents a hydrophobic amino acid such as leucine, isoleucine, valine, or phenylalanine, “N” may include asparagine, serine, threonine, or cysteine, and “x” represents any amino acid. The above repeating units are arranged in succession in two or more numbers, typically ranging from a few to tens, to form a single LRR domain, thereby forming an arc-shaped or horseshoe-shaped three-dimensional structure for the entire domain. Structurally, parts of each repeating unit form β-strands, which are aligned parallel to each other to form a continuous β-sheet on the concave surface of the domain. In contrast, the convex surface consists of α-helices, loops, or other secondary structural elements, exhibiting a more variable structure. Due to these structural features, the LRR domain provides a broad and continuous binding surface, and the concave surface, in particular, serves as a binding site for various ligands or proteins.Therefore, LRR domains play a key role in protein-protein interactions, protein-ligand binding, signal transduction, and molecular recognition processes. Furthermore, LRR domains are widely present across eukaryotes, prokaryotes, and viruses, and are found in proteins involved in various biological processes such as immune responses, cell signaling, cell adhesion, and developmental regulation. For example, innate immunity-related receptors, such as mammalian Toll-like receptors (TLRs) or NOD-like receptors (NLRs), recognize pathogen-derived molecules through LRR domains, while in plants, LRR domains play an important role in pathogen resistance proteins and receptor-like proteins. In some embodiments, the LRR domain may include cap structures (LRRNT, LRRCT) located at the N-terminus and / or C-terminus, which function to increase the stability of the repeat structure and protect the hydrophobic core. Additionally, non-LRR regions (island domains) may be inserted between LRR repeats, which may contribute to specific ligand binding or functional specificity. Therefore, the LRR domain functions as a structural platform that mediates selective interactions with various molecules based on repetitive leucine-rich sequences and unique three-dimensional structures, and can be understood as a protein domain that plays an important role in physiological and pathological processes.

[0014] Metabolic diseases, such as obesity, insulin resistance, type 2 diabetes, and metabolic syndrome, are closely associated with chronic low-grade inflammation, and these inflammatory responses can be mediated by the activation of receptors containing LRR domains. In particular, receptors possessing specific LRR domains have been reported to induce the expression of inflammatory cytokines in adipose tissue, liver, and muscle tissue, thereby inhibiting insulin signaling pathways. In some embodiments, antibodies that specifically bind to an LRR domain can inhibit downstream signaling pathways by blocking ligand binding or receptor activation through said domain. Accordingly, the secretion of inflammatory cytokines (e.g., TNF-α, IL-6, etc.) is reduced, and insulin signaling pathways are restored, which can improve insulin sensitivity. Furthermore, such antibodies can contribute to inhibiting the abnormal expansion and fat accumulation of adipocytes and normalizing energy metabolism. In another embodiment, administration of the antibody can induce improvements in various metabolic indicators, such as inhibition of weight gain, reduction of blood glucose concentration, improvement of glycated hemoglobin (HbA1c) levels, and improvement of blood lipid profiles (e.g., triglycerides, LDL cholesterol, etc.). This can be achieved by selectively regulating inflammatory and metabolic abnormal signaling mediated by the LRR domain. The antibody may be used alone or in combination with existing treatments for metabolic diseases and may be administered via various routes, such as intravenous, subcutaneous, or intramuscular injections. Additionally, the antibody may be provided in the form of a humanized antibody, a chimeric antibody, or an antibody fragment. Accordingly, the antibody binding to the LRR domain can be usefully applied to the prevention and treatment of various metabolic diseases based on a novel mechanism capable of simultaneously regulating inflammatory responses and metabolic regulatory pathways.

[0015] Antibodies that specifically bind to the leucine-rich repeat domain (LRR domain) can be utilized as an effective means for the prevention and / or treatment of various neurodegenerative diseases, including Alzheimer's disease. LRR domains are found in numerous neuroreceptor proteins and are known to play important roles, particularly in extracellular ligand recognition, synapse formation and maintenance, and the regulation of neuroinflammatory responses. For example, Toll-like receptors (TLRs), NOD-like receptors (NLRs), and other LRR-containing proteins containing LRR domains are expressed in microglia and neurons to recognize pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) and mediate subsequent inflammatory signaling. In neurodegenerative diseases such as Alzheimer's disease, amyloid-beta accumulation, hyperphosphorylation and aggregation of tau proteins, and chronic neuroinflammatory responses act as major pathological mechanisms. In particular, receptors containing an LRR domain may recognize pathological proteins such as amyloid-beta or be activated by them to induce the secretion of inflammatory cytokines (e.g., IL-1β, TNF-α, etc.), which can consequently lead to neuronal damage and impaired synaptic function. In some embodiments, an antibody that specifically binds to an LRR domain can regulate excessive neuroinflammatory responses by inhibiting ligand binding or receptor activation through said domain. For example, said antibody may alleviate neuronal damage and exhibit neuroprotective effects by inhibiting microglia hyperactivation and reducing the secretion of inflammatory cytokines. Additionally, said antibody may contribute to reducing the formation and accumulation of amyloid plaques by blocking the recognition or signaling of amyloid-beta.In another embodiment, administration of the antibody inhibits synapse loss and maintains or restores the functional connectivity of neural circuits, thereby mitigating the decline in cognitive functions such as learning and memory abilities. Additionally, it may indirectly affect signaling pathways associated with tau pathology, thereby exhibiting an effect of inhibiting the abnormal phosphorylation and aggregation of tau proteins. The antibody may be designed to cross the blood-brain barrier (BBB) ​​or may be combined with a delivery system that facilitates BBB penetration, and may be administered via intravenous injection, subcutaneous injection, or other appropriate routes. Furthermore, it may be provided in the form of humanized antibodies, chimeric antibodies, or antibody fragments such as Fab or scFv. Thus, antibodies binding to the LRR domain can be usefully applied to the prevention and treatment of various neurological diseases, including Alzheimer's disease, based on a novel mechanism of action capable of simultaneously regulating neuroinflammatory responses and pathological protein accumulation.

[0016] Antibodies that specifically bind to the leucine-rich repeat domain (LRR domain) can be utilized as an effective means for the prevention and / or treatment of fibrosis. The LRR domain is a structural motif found in various receptors and extracellular proteins that plays a crucial role in mediating extracellular matrix (ECM) signaling, the recognition of growth factors and inflammatory ligands, and the subsequent intracellular signaling. In particular, proteins containing the LRR domain are known to be involved in major pathological processes of fibrosis, such as fibroblast activation, differentiation into myofibroblasts, and the excessive accumulation of ECM proteins. Fibrosis can occur in various organs, including the liver, lungs, kidneys, and heart, and is induced by chronic inflammation and tissue damage. In this process, receptors or binding proteins containing an LRR domain interact with transforming growth factor-β (TGF-β), platelet-derived growth factor (PDGF), and other fibrosis-related signaling pathways to promote the proliferation and activation of fibroblasts and induce the excessive production and accumulation of ECM components such as collagen and fibronectin. In some embodiments, an antibody that specifically binds to the LRR domain can inhibit the activation of fibrosis-related signaling pathways by blocking ligand binding or receptor-receptor interactions through said domain. For example, said antibody can inhibit the differentiation of fibroblasts into myofibroblasts by reducing the activation of SMAD proteins acting downstream of the TGF-β signaling pathway or by inhibiting PDGF-mediated signals. Consequently, the expression of α-smooth muscle actin (α-SMA) is reduced, and the expression and accumulation of ECM proteins such as collagen type I and III and fibronectin can be significantly inhibited.In another embodiment, administration of the antibody can inhibit the progression of fibrosis by reducing the secretion of inflammatory cytokines (e.g., TGF-β, IL-1β, TNF-α, etc.) and alleviating inflammatory responses within the tissue. Additionally, it can promote the remodeling of the ECM in already fibrotic tissue and contribute to restoring the normal structure and function of the tissue. The antibody may be used alone or in combination with an antifibrotic agent and may be administered via various routes, such as intravenous, subcutaneous, or intramuscular injection. Furthermore, it may be provided in the form of a humanized antibody, a chimeric antibody, or antibody fragments such as Fab or scFv. Thus, an antibody binding to the LRR domain can serve as a useful therapeutic agent capable of effectively preventing and treating fibrosis in various organs through a novel mechanism that regulates fibroblast activation and ECM accumulation.

[0017] In the present invention, the term “binding molecule” means an intact immunoglobulin comprising a monoclonal antibody, such as a chimeric, humanized, or human monoclonal antibody, or a variable domain comprising an immunoglobulin fragment that competes with the intact immunoglobulin for binding to an antigen, for example, monomeric HA or trimeric HA of the influenza A virus. Regardless of structure, the antigen-binding fragment binds to the same antigen recognized by the intact immunoglobulin. The antigen-binding fragment may comprise a peptide or polypeptide comprising an amino acid sequence of two or more consecutive amino acid residues, 20 or more consecutive amino acid residues, 25 or more consecutive amino acid residues, 30 or more consecutive amino acid residues, 35 or more consecutive amino acid residues, 40 or more consecutive amino acid residues, 50 or more consecutive amino acid residues, 60 or more consecutive amino acid residues, 70 or more consecutive amino acid residues, 80 or more consecutive amino acid residues, 90 or more consecutive amino acid residues, 100 or more consecutive amino acid residues, 125 or more consecutive amino acid residues, 150 or more consecutive amino acid residues, 175 or more consecutive amino acid residues, 200 or more consecutive amino acid residues, or 250 or more consecutive amino acid residues.

[0018] In the present invention, the term “antigen-binding fragment” includes, in particular, Fab, F(ab'), F(ab')2, Fv, dAb, Fd, complementarity determining region (CDR) fragment, single-strand antibody (scFv), bivalent single-strand antibody, single-strand phage antibody, diabody, triabody, tetrabody, polypeptide containing one or more fragments of immunoglobulin sufficient to bind to a specific antigen. The fragment may be produced synthetically, by enzymatic or chemical degradation of complete immunoglobulin, or genetically engineered using recombinant DNA technology. Methods of production are well known in the art.

[0019] In the present invention, the binding molecule may further include an Fc region (Fragment crystallization region) or a constant region. In this case, the Fc region may be the Fc region of an IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, or may be derived therefrom, or may be a hybrid Fc region.

[0020] In the present invention, the Fc region may be the Fc region of a mammalian-derived IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, and preferably may be the Fc region of a human-derived IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, but is not limited thereto.

[0021] As an example of the present invention, the Fc region may be a human-derived immunoglobulin lambda constant region, but is not limited thereto.

[0022] In the present invention, the "hybrid Fc" can be derived from a combination of human IgG subclasses or a combination of human IgD and IgG. When the hybrid Fc binds to a biologically active molecule, polypeptide, etc., it not only increases the serum half-life of the biologically active molecule but also has the effect of increasing the expression level of the polypeptide when a nucleotide encoding an Fc-polypeptide fusion protein is expressed.

[0023] In the bonding molecule of the present invention, the Fc or invariant region may be connected to the variable region by a linker. In this case, the linker is connected to the C-terminus of the Fc or invariant region, and the N-terminus of the bonding molecule of the present invention may be connected to the linker, but is not limited thereto.

[0024] In the present invention, the "linker" may include a sequence that can be cleaved by an enzyme that is overexpressed within the tissue or cell of the target disease. In the case where it can be cleaved by an overexpressed enzyme as described above, the reduction of polypeptide activity due to Fc or the constant region can be effectively prevented. In the present invention, a preferred example of the linker may be a peptide linker composed of 33 amino acids located at positions 282 to 314 of human albumin, which is most abundant in blood, and more preferably a peptide linker composed of 13 amino acids located at positions 292 to 304. These parts are mostly exposed to the outside in terms of the three-dimensional structure and are parts where the possibility of inducing an immune response in the body is minimized. However, the invention is not limited thereto.

[0025] The binding molecule of the present invention is characterized as being an antibody or a fragment thereof, but is not limited thereto. The antibody includes all of the following: a monoclonal antibody, a full-length antibody, or an antibody fragment capable of binding to the Lrig-1 protein as a part of an antibody and capable of binding competitively to the Lrig-1 antigenic determinant site with the binding molecule of the present invention.

[0026] In the present invention, the "antibody" refers to a protein molecule that acts as a receptor for specifically recognizing an antigen, comprising an immunoglobulin molecule that is immunologically reactive with a specific antigen. For the purposes of the present invention, the antigen may be the Lrig-1 protein present on the surface of a regulatory T cell. Preferably, it may specifically recognize the leucine-rich region (LRR) or the immunoglobulin-like domain of the Lrig-1 protein, but is not limited thereto.

[0027] In the present invention, the "immunoglobulin" has a heavy chain and a light chain, and each heavy chain and light chain includes an invariant region and a variable region. The variable regions of the light chain and heavy chain include three variable regions and four framework regions called complementarity determining regions (hereinafter referred to as "CDRs"). The CDRs primarily serve to bind to epitopes of antigens. The CDRs of each chain are typically designated sequentially as CDR1, CDR2, and CDR3 starting from the N-terminus, and are also identified by the chain in which a specific CDR is located.

[0028] In addition, in the present invention, the term "monoclonal antibody" refers to an antibody molecule of a single molecular composition obtained from substantially the same group of antibodies, which exhibits single-binding specificity and affinity for a specific epitope.

[0029] In the present invention, the "full-length antibody" has a structure having two full-length light chains and two full-length heavy chains, each light chain being connected to the heavy chain by a disulfide bond, and includes IgA, IgD, IgE, IgM, and IgG. The IgG includes IgG1, IgG2, IgG3, and IgG4 as subtypes.

[0030] Furthermore, in the present invention, the "antibody fragment" refers to a fragment possessing an antigen-binding function and includes Fab, Fab', F(ab')2, and Fv, etc. Fab has a structure having variable regions of the light and heavy chains, a constant region of the light chain, and a first constant region of the heavy chain (CH1 domain), and has one antigen-binding site. Additionally, Fab' differs from Fab in that it has a hinge region containing one or more cysteine ​​residues at the C-terminus of the heavy chain CH1 domain. The F(ab')2 antibody is generated when the cysteine ​​residues in the hinge region of Fab' form disulfide bonds. Fv (Variable fragment) refers to a minimal antibody fragment possessing only a heavy chain variable region and a light chain variable region. In double-stranded Fv (dsFv), the heavy chain variable region and the light chain variable region are connected by a disulfide bond, and in single-stranded Fv (scFv), the heavy chain variable region and the light chain variable region are generally connected by a covalent bond through a peptide linker. When using a proteolytic enzyme, such as papain or pepsin, the antibody fragment can be a Fab or F(ab')2 fragment, and can be produced through genetic recombination technology.

[0031] In addition, the antibody in the present invention may be a chimeric antibody, a humanized antibody, a bivalent, a bispecific molecule, a minibody, a domain antibody, a bispecific antibody, an antibody mimic, a unibody, a diabody, a triabody, a tetrabody, or a fragment thereof, but is not limited thereto.

[0032] In the present invention, the "chimeric antibody" is an antibody formed by recombining the variable region of a mouse antibody and the constant region of a human antibody, and is an antibody in which the immune response is significantly improved compared to the mouse antibody.

[0033] In addition, in the present invention, the "humanized antibody" refers to an antibody in which the protein sequence of an antibody derived from a non-human species is modified to be similar to a naturally produced antibody variant in humans. For example, the humanized antibody can be produced by recombining a mouse-derived CDR with a human antibody-derived FR to produce a humanized variable region, and then recombining this with a preferred human antibody constant region to produce the humanized antibody.

[0034] In the present invention, the binding molecule may also be provided as a bispecific antibody or a bispecific antigen binding fragment that can bind to an Lrig-1 protein containing an LRR domain and can also bind to other proteins.

[0035] In the present invention, the bispecific antibody and the bispecific antigen-binding fragment may comprise a binding molecule according to the present invention. In one example in the present invention, the bispecific antibody and the bispecific antigen-binding fragment comprise an antigen-binding domain capable of binding to an Lrig-1 protein comprising an LRR domain, wherein the antigen-binding domain capable of binding to an Lrig-1 protein comprising an LRR domain may comprise or be composed of a binding molecule according to the present invention.

[0036] The bispecific antibody and bispecific antigen-binding fragment provided in the present invention comprise an antigen-binding domain, which is a binding molecule capable of binding to an Lrig-1 protein comprising an LRR domain according to the present invention, and an antigen-binding domain capable of binding to another target protein. Here, the antigen-binding domain capable of binding to another target protein may be a protein other than the Lrig-1 protein, and, although not limited thereto, may be, for example, an antigen-binding domain capable of binding to PD-1 or a cell surface receptor.

[0037] The bispecific antibody and bispecific antigen binding fragment according to the present invention may be provided in any suitable format, for example, in the format described in the literature in which the full text is cited by reference herein. For example, bispecific antibodies or bispecific antigen-binding fragments include bispecific antibody conjugates (e.g., IgG2, F(ab')2, or CovX-bodies), bispecific IgG or IgG-type molecules (e.g., IgG, scFv4-Ig, IgG-scFv, scFv-IgG, DVD-Ig, IgG-sVD, sVD-IgG, or 2-in 1-IgG, mAb2, or Tandemab common LC), asymmetric bispecific IgG or IgG-type molecules (e.g., kih IgG, kih IgG common LC, CrossMab, kih IgG-scFab, mAb-Fv, charge pairs, or SEED-bodies), and small bispecific antibody molecules (e.g., diabody (Db), dsDb, DART, scDb, tandAbs, tandem scFv (taFv), tandem dAb / VHH, triple It may be a body, triple head, Fab-scFv, or F(ab')2-scFv2), a bispecific Fc and CH3 fusion protein (e.g., taFv-Fc, di-diabody, scDb-CH3, scFv-Fc-scFv, HCAb-VHH, scFv-kih-Fc, or scFv-kih-CH3), or a bispecific fusion protein (e.g., scFv2-albumin, scDb-albumin, taFv-toxin, DNL-Fab3, DNL-Fab4-IgG, DNL-Fab4-IgG-cytokine2). Those skilled in the art can design and manufacture a bispecific antibody and a bispecific antigen-binding fragment according to the present invention.

[0038] The method for producing the bispecific antibody in the present invention comprises chemically crosslinking an antibody or antibody fragment with a reducing disulfide or non-reducing thioether bond. For example, N-succinimidyl-3-(-2-pyridyldithio)-propionate (SPDP) may be used to chemically crosslink a Fab fragment, for example, through a hinge region SH- group, to produce a disulfide-linked bispecific F(ab)2 heterodimer.

[0039] In addition, another method for producing the bispecific antibody in the present invention comprises fusing an antibody-producing hybridoma with, for example, polyethylene glycol to produce a quadroma cell capable of secreting the bispecific antibody.

[0040] The bispecific antibody and bispecific antigen-binding fragment according to the present invention can be produced by recombination, for example, by expression from a nucleic acid construct encoding a polypeptide for an antigen-binding molecule.

[0041] For example, a DNA construct comprising a sequence encoding light chain and heavy chain variable domains for two antigen-binding domains (i.e., a light chain and heavy chain variable domain for an antigen-binding domain capable of binding to PD-1, etc., and a light chain and heavy chain variable domain for an antigen-binding domain capable of binding to other target proteins) and encoding a suitable linker or dimerization domain between the antigen-binding domains can be produced by molecular cloning technology. The recombinant bispecific antibody can then be produced by expression of the construct (e.g., in vitro) in a suitable host cell (e.g., mammalian host cell), and the expressed recombinant bispecific antibody can then be optionally purified.

[0042] Antibodies can be produced by an affinity maturation process in which modified antibodies are produced in which the affinity of the antibody for the antigen is improved compared to the unmodified parent antibody. Affinity-matured antibodies can be produced by procedures known in the art.

[0043] In addition, the binding molecule provided in the present invention may include variants of the amino acid sequence as long as they can specifically bind to the Lrig-1 protein. For example, the amino acid sequence of the antibody may be modified to improve the binding affinity and / or other biological properties of the antibody. Such modifications include, for example, deletion, insertion, and / or substitution of amino acid sequence residues of the antibody.

[0044] These amino acid variations are based on the relative similarities of amino acid side chain substituents, such as hydrophobicity, hydrophilicity, charge, size, etc. Analysis of the size, shape, and type of amino acid side chain substituents reveals that arginine, lysine, and histidine are all positively charged residues; alanine, glycine, and serine have similar sizes; and phenylalanine, tryptophan, and tyrosine have similar shapes. Therefore, based on these considerations, arginine, lysine, and histidine; alanine, glycine, and serine; and phenylalanine, tryptophan, and tyrosine can be considered biologically functional equivalents.

[0045] In introducing mutations, the hydropathic index of the amino acids may be considered. Each amino acid is assigned a hydropathic index based on its hydrophobicity and charge: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine / cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5). The hydrophobic amino acid index is very important in conferring interactive biological functions of proteins. It is a known fact that similar biological activity can be achieved by substituting with amino acids having similar hydrophobic indices. When introducing a variation based on the hydrophobic index, the substitution is preferably made between amino acids exhibiting a difference in hydrophobic index within ±2, more preferably within ±1, and even more preferably within ±0.5.

[0046] Meanwhile, it is also well known that substitution between amino acids having similar hydrophilicity values ​​results in proteins with uniform biological activity. As already known, the following hydrophilicity values ​​are assigned to each amino acid residue: arginine (+3.0); lysine (+3.0); aspalate (+3.0 ± 1); glutamate (+3.0 ± 1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5 ± 1); alanine (-0.5); histidine (-0.5); cysteine ​​(-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); tryptophan (-3.4). When introducing a variation by referring to the hydrophilicity value, substitution can be performed between amino acids exhibiting a difference in hydrophilicity value within preferably ± 2, more preferably ± 1, and even more preferably ± 0.5.

[0047] Amino acid exchanges in proteins that do not alter the overall activity of the molecule are known in the art. The most common exchanges are those between amino acid residues Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, and Gln / Glu.

[0048] Considering the variant having the aforementioned biological equivalent activity, the binding molecule of the present invention is interpreted to include a sequence that exhibits substantial identity with the sequence listed in the sequence list.

[0049] In the present invention, the term "substantial identity" refers to a sequence in which, when the sequence of the present invention is paralleled with any other sequence to correspond as much as possible and the parallel sequence is analyzed using an algorithm commonly used in the art, it exhibits at least 61% homology, more preferably 70% homology, even more preferably 80% homology, and most preferably 90% homology. Alignment methods for sequence comparison are known in the art. Various methods and algorithms for alignment are accessible from the NCBI Basic Local Alignment Search Tool (BLAST), NBCI (National Center for Biological Information), etc., and can be used in conjunction with sequence analysis programs such as blastp, blasm, blastx, tblastn, and tblastx on the internet. BLAST is accessible at this address (www.ncbi.nlm.nih.gov / BLAST / ). Methods for comparing sequence homology using this program can be found online (www.ncbi.nlm.nih.gov / BLAST / blast_help.html).

[0050] In the present invention, the binding molecule, preferably the antibody, can be produced by a conventional method of producing antibodies, but can also be produced by affinity maturation.

[0051] In the present invention, "affinity maturation" refers to the process in which activated B cells produce antibodies with increased affinity for an antigen during an immune response. For the purposes of the present invention, the affinity maturation can produce antibodies or antibody fragments generated by affinity maturation based on the principles of mutation and selection, just as processes occur in nature.

[0052] According to another embodiment of the present invention, a nucleic acid molecule encoding the binding molecule provided in the present invention is provided.

[0053] The nucleic acid molecules of the present invention include all nucleic acid molecules in which the amino acid sequence of the binding molecule provided in the present invention is translated into a polynucleotide sequence as is known to those skilled in the art. Therefore, various polynucleotide sequences can be prepared by an ORF (open reading frame), and all of these are also included in the nucleic acid molecules of the present invention.

[0054] According to another embodiment of the present invention, an expression vector is provided in which the isolated nucleic acid molecule provided in the present invention is inserted.

[0055] In the present invention, the "vector" is a nucleic acid molecule capable of transporting another nucleic acid to which a certain nucleic acid molecule is connected. One type of vector is a "plasmid," which refers to circular double-stranded DNA to which additional DNA segments can be ligated. Another type of vector is a phage vector. Another type of vector is a viral vector to which additional DNA segments can be ligated to the viral genome. Some vectors can replicate autonomously in the host cell to which they are introduced (e.g., bacterial vectors are episomal mammalian vectors with a bacterial replication origin). Other vectors (e.g., non-episosomal mammalian vectors) can be incorporated into the host cell's genome upon introduction into the host cell and thereby replicate along with the host genome. Furthermore, some vectors can direct the expression of genes to which they are connected at the operational level. Such vectors are referred herein as "recombinant expression vectors" or simply "expression vectors." In general, expression vectors useful in recombinant DNA techniques often exist in the form of plasmids. In this specification, "plasmid" and "vector" may be used interchangeably because plasmid is the most commonly used form of vector.

[0056] Specific examples of the expression vector in the present invention may be selected from the group consisting of commercially widely used pCDNA vectors, F, R1, RP1, Col, pBR322, ToL, and Ti vectors; cosmids; phages such as lambda, lambdoid, M13, Mu, p1 P22, Qμμ, T-even, T2, T3, and T7; and plant viruses, but are not limited thereto. Any expression vector known to those skilled in the art as an expression vector may be used in the present invention, and the selection of the expression vector depends on the properties of the target host cell. When introducing the vector into the host cell, it may be performed by calcium phosphate transfection, viral infection, DEAE-dextran regulated transfection, lipofectamine transfection, or electroporation, but is not limited thereto. Those skilled in the art may select and use an introduction method suitable for the expression vector and the host cell to be used. Preferably, the vector contains one or more screening markers, but is not limited thereto. Screening is possible based on whether a product is produced using a vector that does not contain screening markers. The selection of screening markers is performed by the target host cells, and since this utilizes methods already known to those skilled in the art, the present invention is not limited thereto.

[0057] To facilitate the purification of the nucleic acid molecule of the present invention, a tag sequence may be inserted into an expression vector and fused. The tag includes, but is not limited to, a hexahistidine tag, a hemagglutinin tag, a myc tag, or a flag tag, and any tag known to those skilled in the art that facilitates purification may be used in the present invention.

[0058] According to another embodiment of the present invention, a host cell line transformed by the expression vector provided in the present invention is provided.

[0059] In the present invention, the "host cell" includes an individual cell or cell culture that may or was a recipient of a vector(s) for the incorporation of a polypeptide insert. The host cell includes progeny of a single host cell, and said progeny may not necessarily be completely identical to the original parent cell (morphologically or in genomic DNA complements) due to natural, accidental, or intentional mutations. The host cell includes a cell that has been transfected in vivo with the polypeptide(s) of the present invention.

[0060] In the present invention, the host cell may include cells of mammalian, plant, insect, fungal, or cellular origin, such as, for example, bacterial cells such as Escherichia coli, Streptomyces, and Salmonella typhimurium; fungal cells such as yeast cells and Pichia pasteoris; insect cells such as Drozophylla and Spodoptera Sf9 cells; animal cells such as CHO (Chinese hamster ovary cells), SP2 / 0 (mice myeloma), human lymphoblastoid, COS, NSO (mice myeloma), 293T, Bow melanoma cells, HT-1080, BHK (baby hamster kidney cells), HEK (human embryonic kidney cells) or PERC.6 (human retinal cells); or plant cells, but is not limited thereto, and any cell that can be used as a host cell line known to those skilled in the art may be used.

[0061] According to another embodiment of the present invention, an antibody-drug conjugate (ADC) comprising an antibody and a drug provided in the present invention is provided.

[0062] In the present invention, the term "Antibody-Drug Conjugate (ADC)" refers to a form in which a drug and an antibody are chemically linked without reducing the biological activity of the antibody and the drug. In the present invention, the antibody-drug conjugate refers to a form in which a drug is bound to an amino acid residue at the N-terminus of the heavy chain and / or light chain of an antibody, specifically, a form in which a drug is bound to an α-amine group at the N-terminus of the heavy chain and / or light chain of an antibody.

[0063] In the present invention, the term "drug" may refer to any substance having specific biological activity in cells, and this concept includes DNA, RNA, or peptides. The drug may be in a form containing a reactive group capable of reacting with an α-amine group to form a crosslink, and may also be in a form in which a linker containing a reactive group capable of reacting with an α-amine group to form a crosslink is connected.

[0064] Examples of reaction groups capable of reacting with the α-amine group to crosslink in the present invention include, as long as they can react with the α-amine group at the N-terminus of the heavy or light chain of an antibody to crosslink, the type thereof is not particularly limited and includes all types known in the art that react with amine groups. Examples may be any one of isothiocyanate, isocyanates, acyl azide, NHS ester, sulfonyl chloride, aldehyde, glyoxal, epoxide, oxirane, carbonate, aryl halide, imidoester, carbodiimide, anhydride, and fluorophenyl ester, but are not limited thereto.

[0065] In the present invention, the drug may be included regardless of its type, as long as it is a drug capable of treating fibrosis, brain and nervous system diseases, or metabolic diseases.

[0066] According to another embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of fibrosis, neurological diseases or metabolic diseases is provided, comprising a chimeric antigen receptor (CAR) as an active ingredient, the chimeric antigen receptor comprising an antigen-specific binding domain, a linking domain, and a CD3 zeta (ζ) signaling domain.

[0067] In the present invention, the term "chimeric antigen receptor" or "CAR" refers to an engineered receptor comprising an extracellular antigen binding domain and an intracellular signaling domain. While the most common type of CAR includes a short-chain variable fragment (scFv) derived from a monoclonal antibody fused to a transmembrane and intracellular domain of a T cell co-receptor, such as the CD3 zeta (ζ) chain, the present invention as described herein is not limited to these domains. Rather, "chimeric antigen receptor" or "CAR" as used herein refers to any receptor engineered to express any intracellular signaling molecule and an extracellular antigen binding domain fused to or linked thereto. In the present invention, said binding domain may include a short-chain variable fragment (scFv) capable of specifically recognizing the Lrig-1 protein. In the present invention, said "short-chain variable fragment" or "scFv" refers to a fusion protein of the variable heavy chain (VH) and variable light chain (VL) of an antibody formed by a peptide linker between VL and VH.

[0068] In addition, in the present invention, the VH domain and the VL domain may be connected through a flexible linker. In the present invention, the flexible linker may be a glycine / serine linker of about 10 to 30 amino acids (e.g., 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 amino acids), and preferably may be 15 amino acid lengths. In the present invention, the linker length can act as an important determining site for the chimeric antigen receptor, so a linker shorter than the above range can increase affinity but can also impair CAR expression by causing intracellular multimer formation, whereas a linker longer than the above range can decrease antigen affinity by moving VL and VH CDR further in space.

[0069] The chimeric antigen receptor of the present invention may further include at least one of a hinge region (or spacer) and a signal transduction domain. In the present invention, the hinge region is a part connecting the antigen binding domain and the transmembrane domain, also called a 'spacer,' and is intended to extend the antigen binding domain from the T cell membrane or NK cell membrane. In the present invention, the hinge region may be obtained from any suitable sequence from any genus including, for example, human or part thereof, or may include, but is not limited to, a hinge region of a human protein including, but not limited to, CD8, CD28, 4-1BB, OX40, all or part of the CD3 zeta (ζ) chain, T cell receptor α or β chain, CD28, CD3ε, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, ICOS, CD154, functional derivatives thereof, or combinations thereof. Additionally, in the present invention, the hinge region may include one selected from immunoglobulins (e.g., IgG1, IgG2, IgG3, IgG4, and IgD) without being limited to, but is not limited to.

[0070] In the present invention, the signaling domain refers to a portion of a chimeric antigen receptor that is found or engineered to be found inside a T cell. In the present invention, the signaling domain may or may not include a transmembrane domain that serves to anchor the chimeric antigen receptor to the plasma membrane of the T cell. In the present invention, the transmembrane domain and the signaling domain may be derived from the same protein (e.g., CD3 zeta(ζ) molecule), or the transmembrane domain and the signaling domain may be derived from different proteins (e.g., the transmembrane domain of CD28 and the intracellular signaling domain of the CD3 zeta(ζ) molecule, or vice versa).

[0071] In the present invention, the transmembrane domain may include, for example, a T cell receptor α or β chain, all or part of a CD3 zeta (ζ) chain, CD28, CD3ε, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, ICOS, CD154, functional derivatives thereof, or combinations thereof, but is not limited thereto. In the present invention, the co-stimulatory domain is 4-1BB (CD137); OX40; CD27; CD28; CD30; CD40; PD-1; CD2; CD7; CD258; Natural Killer Group 2 member C (NKG2C); Natural Killer Group 2 member (NKG2D); B7-H3; CD83; ICAM-1; It may include, but is not limited to, a functional signaling domain derived from a polypeptide comprising a ligand that binds to LFA-1 (CD11a / CD18) or ICOS; an active fragment thereof; a functional derivative thereof; or a combination thereof.

[0072] In the present invention, the signal transduction domain may include, but is not limited to, a functional signal transduction domain derived from a polypeptide comprising all or part of CD3 zeta (ζ), common FcR gamma (FcER1G), Fc gamma RIIIa, Fc R beta (Fc epsilon rib), CD3 gamma, CD3 delta, CD3 epsilon, CD79a, CD79b, DNAX-activated protein 10 (DAP10), DNAX-activated protein 12 (DAP12), an active fragment thereof, a functional derivative thereof, or a combination thereof, and such signal transduction domains are known in the art.

[0073] According to another embodiment of the present invention, the invention relates to a composition for the prevention, improvement, or treatment of fibrosis, neurological diseases, or metabolic diseases comprising, as an active ingredient, a binding molecule of the present invention; a nucleic acid molecule; an expression vector; a host cell line; or an antibody-drug conjugate (ADC).

[0074] The neurological disease to be prevented, improved, or treated by the composition provided in the present invention may be a neurodegenerative disease or a neuroinflammatory disease.

[0075] In the present invention, the "neurodegenerative disease" may refer to a disease caused by a decrease or loss of function of nerve cells, and the "neuroinflammatory disease" may refer to a disease caused by an excessive inflammatory response of the nervous system. Specific examples of the neurodegenerative disease or neuroinflammatory disease in the present invention may be selected from the group consisting of stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Niemann-Pick disease, multiple sclerosis, prion disease, Creutzfeldt-Jakob disease, frontotemporal dementia, Lewy dementia, amyotrophic lateral sclerosis, paraneoplastic syndrome, corticobasal degeneration, multiple system atrophy, progressive supranuclear palsy, neurological autoimmune disease, spinocerebellar ataxia, inflammatory and neuropathic pain, cerebrovascular disease, spinal cord injury, and tauopathy, but It is not limited.

[0076] In another example of the present invention, the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow, and skin, but is not limited thereto. Additionally, the fibrosis includes pulmonary fibrosis, liver fibrosis, cardiac fibrosis, cerebrofibrosis, cutaneous fibrosis, pancreatic fibrosis, bone fibrosis, etc., but is not limited thereto.

[0077] In the present invention, the term "fibrosis" refers to the occurrence of excessive fibrous connective tissue within an organ or tissue. Fibrosis is characterized by the accumulation and remodeling of the extracellular matrix (ECM). Although there are clear etiological and clinical differences that distinguish it from cancer, which is a mass formed by such abnormal cell growth, most chronic fibrotic disorders share a common continuous stimulus that sustains the production of growth factors, proteases, angiogenic factors, and fibrogenic cytokines. Together, these factors stimulate the deposition of connective tissue components, particularly collagen and proteoglycans, thereby continuously remodeling and destroying normal tissue structures.

[0078] In the present invention, 'metabolic disease' refers to a condition or disease that is closely associated with or caused by obesity, and specifically, it may be one or more selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, and arteriosclerosis.

[0079] In this invention, "obesity" refers not merely to having a high body weight, but to a state in which body fat is excessively accumulated. This means that even if a person appears to have a normal weight on the outside, they can be classified as obese if their body fat percentage is high. Obesity is typically assessed using the Body Mass Index (BMI); a BMI of 23–24.9 is classified as overweight, 25–29.9 as mild obesity, 30–34.9 as moderate obesity, and 35 or higher as severe obesity. Obesity occurs due to the complex interplay of multiple factors rather than a single cause, including poor dietary habits (including Westernized eating habits), reduced physical activity, emotional factors, and genetic factors. Consequently, obesity increases the risk of developing chronic diseases such as hyperlipidemia, diabetes, and hypertension.

[0080] In this invention, fatty liver refers to a condition or disease in which fat accumulates in excessive amounts in liver cells due to a disorder of lipid metabolism in the liver.

[0081] In this invention, hyperlipidemia refers to a condition or disease in which the concentration of lipid components in the blood, particularly cholesterol and triglycerides, is higher than normal levels, and is used in a broad sense to include all conditions in which a reduction in blood lipid concentration is required.

[0082] In this specification, arteriosclerosis refers to a condition or disease in which blood circulation to organs and tissues in the body is reduced due to the thickening of artery walls and a decrease in elasticity, and includes the meaning of "atherosclerosis," which refers to a condition or disease in which blood circulation is reduced as the lumen narrows due to the deposition of fat, cholesterol, and other substances on the inner wall of the artery forming plaque. Arteriosclerosis can occur in any part of the body; if it occurs in the blood vessels of the heart, it can cause coronary artery diseases such as angina pectoris and myocardial infarction; if it occurs in the brain, it can cause cerebral infarction; and if it occurs in the kidneys, it can cause renal failure.

[0083] In addition, the composition provided in the present invention may be used as a pharmaceutical composition or a food composition, but is not limited thereto.

[0084] The "prevention" of the present invention may include, without limitation, any act that can block, suppress, or delay symptoms caused by fibrosis, neurological diseases, or metabolic diseases using the composition of the present invention.

[0085] The "treatment" and "improvement" of the present invention may include, without limitation, any act that enables the improvement or benefit of symptoms caused by fibrosis, neurological diseases, or metabolic diseases using the composition of the present invention.

[0086] In the present invention, the pharmaceutical composition may be characterized in that it is in the form of a capsule, tablet, granule, injection, ointment, powder, or beverage, and the pharmaceutical composition may be characterized in that it is intended for humans.

[0087] The pharmaceutical composition of the present invention is not limited to these, but may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as topical preparations, suppositories, and sterile injectable solutions, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. For oral administration, the pharmaceutically acceptable carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. For injectable preparations, it may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc., in combination; and for topical administration, it may include bases, excipients, lubricants, preservatives, etc. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injectables, it can be manufactured in the form of unit dosing ampoules or multiple dosing ampoules. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release formulation, etc.

[0088] Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. Additionally, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be additionally included.

[0089] The routes of administration of the pharmaceutical composition according to the present invention are not limited to but include oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal. Oral or parenteral administration is preferred.

[0090] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0091] The pharmaceutical composition of the present invention may vary depending on several factors including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art, depending on the patient's condition, body weight, degree of disease, drug form, route of administration, and duration, and may be administered at a dose of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be administered once a day or divided into several doses. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, coated tablet, capsule, liquid, gel, syrup, slurry, or suspension.

[0092] A food composition containing the composition of the present invention as an active ingredient can be manufactured in the form of various food products, such as beverages, chewing gum, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, bread, etc. Since the food composition of the present invention is composed of plant extracts that have almost no toxicity or side effects, it can be used safely even when taken for a long period for preventive purposes.

[0093] When the composition of the present invention is included in a food composition, the amount may be added in a ratio of 0.1 to 50% of the total weight.

[0094] Here, when the above food composition is prepared in the form of a beverage, there are no special limitations other than containing the above food composition in the indicated proportions, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in ordinary beverages. That is, as natural carbohydrates, it may include monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, conventional sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. Examples of the above flavoring agents include natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.).

[0095] In addition, the food composition of the present invention may contain various nutritional agents, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0096] These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected in the range of 0.1 to about 50 parts by weight per 100 parts by weight of the composition of the present invention.

[0097]

[0098] The LRR domain functions as a structural platform mediating selective interactions with various molecules based on repetitive leucine-rich sequences and unique stereostructures, and because it is a protein domain that plays an important role in physiological and pathological processes, it can be used as a novel target for therapeutic agents for various metabolic diseases, neurological diseases, and fibrosis. Accordingly, antibodies or antigen-binding fragments that specifically bind to the LRR domain provided in the present invention can prevent or treat various metabolic diseases, neurological diseases, and fibrosis.

[0099]

[0100] Figure 1 is the experimental protocol.

[0101] Figure 2 shows the kinetics of Lrig-1 expression.

[0102] Figure 3 shows the results of confirming whether 3T3-L1 cells differentiated.

[0103] Figure 4a shows the results of confirming whether 3T3-L1 cells are differentiated using a 100x microscope.

[0104] Figure 4b shows the results of confirming whether 3T3-L1 cells are differentiated using a 400x microscope.

[0105] Figure 5 shows the results of confirming the change in body weight of Lrig-1 knockout mice for females and males.

[0106] Figure 6 shows the results of checking the body weight change of the first confirmed Lrig-1 knockout male mice, divided into cases where the genotype was heterozygous or homozygous.

[0107] Figure 7 shows the results of examining the changes in adipocytes of the first Lrig-1 knockout male mouse, divided into visceral fat and subcutaneous fat. In the figure, PGF represents eWAT, i.e., visceral fat, and SCF represents iWAT, i.e., subcutaneous fat.

[0108] Figure 8 shows the results of checking the body weight change of the first confirmed Lrig-1 knockout female mouse, divided into cases where the genotype is heterozygous or homozygous.

[0109] Figure 9 shows the results of examining the changes in adipocytes of the first Lrig-1 knockout female mouse, divided into visceral fat and subcutaneous fat. In the figure, PGF represents eWAT, i.e., visceral fat, and SCF represents iWAT, i.e., subcutaneous fat.

[0110] Figure 10 shows the results of confirming the change in body weight of the second Lrig-1 knockout male mouse, divided into cases where the genotype was heterozygous or homozygous.

[0111] Figure 11 shows the results of examining changes in adipocytes of Lrig-1 knockout male mice, divided into visceral fat and subcutaneous fat. In the figure, PGF represents eWAT, i.e., visceral fat, and SCF represents iWAT, i.e., subcutaneous fat; WAT represents white adipose tissue, i.e., white fat cells; and BAT represents brown adipose tissue, i.e., brown fat cells.

[0112] Figure 12 shows the results of confirming the change in body weight of the second Lrig-1 knockout female mouse, divided into cases where the genotype was heterozygous or homozygous.

[0113] Figure 13 shows the results of examining changes in adipocytes of Lrig-1 knockout female mice, divided into visceral fat and subcutaneous fat. In the figure, PGF represents eWAT, i.e., visceral fat, and SCF represents iWAT, i.e., subcutaneous fat.

[0114] Figure 14 illustrates the experimental process of differentiating 3T3 cell lines to express Lrig-1.

[0115] Figure 15 shows the expression level of Lrig-1 confirmed through flow cytometry analysis.

[0116] Figure 16 shows the degree of expression of Lrig-1 over time.

[0117] Figure 17 is data on the obesity treatment effect of active ingredients, such as antibodies targeting the LRR domain identified in the present invention.

[0118] Figure 18 is data on the dementia treatment effect of an active ingredient, such as an antibody targeting the LRR domain, identified in the present invention.

[0119] Figure 19 is data on the therapeutic effect of active ingredients, such as antibodies targeting the LRR domain identified in the present invention, on lung and liver fibrosis.

[0120] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the invention.

[0121]

[0122] [Example 1] Lrig-1 knockdown in 3T3-L1

[0123] [Example 1-1] Induction for 3T3-L1 differentiation

[0124] First, 1 x 10⁶ 3T3-L1 cells (pre-adipocytes) were placed in a 24-well plate. 5Dispense into / wells and incubate for 3 days in DMI medium (medium containing 10% fetal bovine serum of Cytiva® Cat. No. SV30207.02, 1% penicillin-streptomycin of Thermo Fisher® Cat. No. 15140122, 1 µM dexamethasone of Sigma® Cat. No. D4902, 520 µM 3-isobutyl-1-methylxanthine (IBMX) of Sigma® Cat. No. 228420010, and 1 µM insulin of Sigma® Cat. No. I5500 in Dulbecco's modified Eagle's medium (DMEM) for Cytiva® Cat. No. SH30243.01, 1% penicillin-streptomycin of Thermo Fisher® Cat. No. 15140122, 1 µM dexamethasone of Sigma® Cat. No. D4902, 520 µM 3-isobutyl-1-methylxanthine (IBMX) of Sigma® Cat. No. 228420010, and 1 µM insulin of Sigma® Cat. No. I5500), then in insulin medium (Cultured for 3 days in Dulbecco's modified Eagle's medium (DMEM) of Cytiva® Cat. No. SH30243.01 containing 10% fetal bovine serum of Cytiva® Cat. No. SV30207.02, 1% penicillin-streptomycin of Thermo Fisher® Cat. No. 15140122, and 167 nM insulin of Sigma® Cat. No. I5500.) Subsequently, the cultures were incubated for 6 days in complete DMEM medium (Dulbecco's modified Eagle's medium (DMEM) of Cytiva® Cat. No. SH30243.01 containing 10% fetal bovine serum of Cytiva® Cat. No. SV30207.02 and 1% penicillin-streptomycin of Thermo Fisher® Cat. No. 15140122). A detailed experimental protocol is shown in Figure 1, and the medium conditions are shown in Table 1 below.

[0125] Reagents | Concentration | Catalog No. Completed DMEM (DMEM) | Dulbecco's modified Eagle's medium (DMEM) Cytiva, SH3024 3.01 Fetal bovine serum 10% Cytiva, SV3020 7.02 Penicillin-Streptomycin 1% Thermo Fisher, 15140122 Maintain media (MM) | Dulbecco's modified Eagle's medium (DMEM) Cytiva, SH3024 3.01 Bovine calf serum 10% Cytiva, SH3007 3.03 HI Penicillin-Streptomycin 1% Thermo Fisher, 15140122 DMID | Dulbecco's modified Eagle's medium (DMEM) Cytiva, SH30243.01 Fetal bovine serum 10% Cytiva, SV30207.02 Penicillin-Streptomycin 1% Thermo Fisher, 15140122 Deximethasone (Dex) 1 μMSigma, D49023-isobutyl-1-methylxanthine (IBMX) 520 μMSigma, 228420010 Insulin (I) 1 μMSigma, I5500 Insulin media Dulbecco's modified Eagle's medium (DMEM) Cytiva, SH30243.01 Fetal bovine serum 10% Cytiva, SV30207.02 Penicillin-Streptomycin 1% Thermo Fisher, 15140122 Insulin (Insulin (I)) 167 nMSigma, I5500 FACS analysis Mouse LRIG1 Alexa 488-conjugated Antibody R&D systems, FAB3688 GBODIPY™ 493 / 503 (4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene) 1 μM nvitrogen, D3922.

[0126] [Examples 1-2] siRNA transfection

[0127] 1 x 10 3T3-L1 cells induced in Example 1-1 5 After 3 days of seeding into cells / well, siRNA transfection was performed at 100% density and in a state of cell cycle arrest. The control group and Lrig-1 siRNA were prepared according to Table 2 below.

[0128] siRNAStock Conc.Working Conc.Media)siRNA-Control (AM1611, Thermo Fisher)5µM240nMOpti-MEMsiRNA-Lrig1 (s68320, Thermo Fisher)10µM240nMOpti-MEM

[0129] Next, RANiMAX (Stock 1 µL / Opti-MEM 50 µL) was prepared, and 100 µL of siRNA mixture was prepared by mixing 50 µL of RANiMAX with 50 µL of the siRNA prepared in Table 2. The 3T3-L1 cells prepared in Example 1-1 were washed twice with S / F DMEM, followed by treatment with 500 µL of S / F DMEM. Subsequently, 100 µL of the siRNA mixture was applied to each of the negative control, control, and siRNA-treated groups (total 600 µL, final concentration 20 nM, 24 h o / n). Adipocyte differentiation was then induced.

[0130]

[0131] [Example 2] Confirmation of Lrig-1 expression level after knockdown

[0132] The expression level of Lrig-1 protein was analyzed by FACS on the 5th day after knockdown and the 4th day after induction. Specifically, the expression level of Lrig-1 protein was analyzed by FACS using the Mouse LRIG1 Alexa488-conjugated Antibody from R&D systems® Cat. No. FAB3688G.

[0133] As a result, it was confirmed that the Lrig-1 protein was knocked down in the group treated with siRNA (not shown).

[0134]

[0135] [Example 3] Comparison of Lrig-1 expression and terminal differentiation

[0136] [Example 3-1] Confirmation of Inhibition of Differentiation into Adipocytes Upon Lrig-1 Knockdown

[0137] The expression level of Lrig-1 protein was analyzed via FACS on the 13th day after knockdown and on the 12th day after induction. Specifically, the expression of Lrig-1 protein was confirmed using the Mouse LRIG1 Alexa488-conjugated Antibody of R&D systems® Cat. No. FAB3688G, and 1 µM of BODIPY™ 493 / 503 (4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene) of Invitrogen® Cat. No. D3922 was used to confirm via FACS whether pre-adipocytes had differentiated into adipocytes.

[0138] As a result, it was confirmed that Lrig-1 protein was knocked down in the group treated with siRNA, and that the differentiation of pre-adipocytes into adipocytes was inhibited compared to the control group (not shown).

[0139]

[0140] [Example 3-2] Confirmation of Inhibition of Differentiation into Adipocytes Upon Lrig-1 siRNA Treatment

[0141] Previously, the inhibition of differentiation into adipocytes confirmed in Example 3-1 was specifically confirmed in terms of Lrig-1 expression kinetics and lipid droplet 3T3-L1 differentiation.

[0142] As a result, as shown in Figure 2, it was confirmed that when Lrig-1 expression was knocked down in 3T3-L1 cells by treatment with Lrig-1 siRNA, the expression of Lrig-1 in adipocytes and preadipocytes was inhibited, and as shown in Figure 3, it was confirmed that the formation of lipid droplets was inhibited in 3T3-L1 cells treated with Lrig-1 siRNA.

[0143] In addition, as shown in Figures 4a and 4b, microscopic images confirmed that the formation of lipid droplets in 3T3-L1 cells was inhibited in Lrig-1 knockdown cells.

[0144]

[0145] [Example 4] Analysis of Treg Lrig-1 knockout mouse experimental results

[0146] [Example 4-1] Preparation of L1 wKO(Lrig-1 CreERT2) mouse

[0147] L1 wKO(Lrig-1 CreERT2) mouse is a whole-body knockout mouse of Lrig1 that can knock out Lrig1 by inserting the creERT2 sequence into the translational initiation site of the endogenouse Lrig1 locus and report Lrig1 expression as creERT2.

[0148]

[0149] [Example 4-2] Observation of body weight changes in L1 wKO (Lrig-1 CreERT2) mice

[0150] The L1 wKO (Lrig-1 CreERT2) mice prepared in Example 4-1 were divided into homo, hetero knockout mice, females, and males, and changes in body weight were observed for 6 to 8 weeks.

[0151] Changes in genotype and body weight of L1 wKO(Lrig-1 CreERT2) mice are shown in Table 3 and Figure 5 below.

[0152]

[0153]

[0154] As a result, it was found that among L1 wKO (Lrig-1 CreERT2) mice, homo knockout mice had a body weight reduced by 14.7% to 22.4% compared to hetero knockout mice.

[0155]

[0156] [Example 4-3] Confirmation of adipose tissue reduction and body weight reduction in L1 wKO (Lrig-1 CreERT2) mice

[0157] Changes in the weight of adipose tissue and body weight of L1 wKO (Lrig-1 CreERT2) mice were examined twice. Specifically, changes in adipose tissue and body weight of male and female mice were examined over 5 to 12 weeks and are shown in Figures 6 to 9.

[0158] As a result, in homo knockout mice, males showed a 12.51% decrease in body weight compared to the control group at week 12, and visceral and subcutaneous fat also decreased; in females, body weight also decreased by 14.82% compared to the control group at week 12, and visceral and subcutaneous fat also decreased.

[0159]

[0160] Changes in fat tissue and body weight of male and female mice were checked once again for 6 to 12 weeks and are shown in Figures 10 to 13.

[0161] As a result, in homo knockout mice, males showed an 8.98% decrease in body weight compared to the control group at week 12, and it was observed that visceral and subcutaneous fat also decreased; additionally, a decrease in brown fat cells was confirmed. In females as well, body weight decreased by 14.84% compared to the control group at week 12, and it was observed that visceral and subcutaneous fat also decreased.

[0162]

[0163] Thus, it was confirmed that inhibiting the function of Treg L1 reduces fat cells and body weight, thereby demonstrating its therapeutic effect on metabolic diseases, including obesity.

[0164]

[0165] [Example 5] Diagnosis of metabolic disease through confirmation of Lrig-1 increase in adipocytes

[0166] [Example 5-1] Differentiation of 3T3-L1 cells into adipocytes

[0167] 3T3-L1 cell lines, which are pre-adipocyte cell lines expressing Lrig-1, were placed in a 24-well plate at a ratio of 1 x 10⁶ 5 Distilled into wells and cultured in DMI medium for two days, followed by insulin medium for two days, and then DMEM medium for two days.

[0168] The specific experimental protocol is shown in Table 4 and Figure 14 below.

[0169]

[0170]

[0171] [Example 5-2] Analysis of Lrig-1 expression levels in differentiated adipocytes

[0172] In this study, flow cytometry was performed using a mouse Lrig-1 Alexa488-binding antibody to analyze the level of Lrig-1 expression in differentiated adipocytes.

[0173] The results are shown in Figures 15 and 16, and it was found that the expression of Lrig1-1 was highest on day 4, which allows for the diagnosis of metabolic diseases including obesity.

[0174]

[0175] [Example 6] Confirmation of therapeutic effects for obesity, dementia, and osteoporosis

[0176] 1. Preparation of antibodies, etc. that specifically bind to the LRR domain

[0177] As for previously known active ingredients, antibodies that specifically bind to the LRR domain and LRRC4 family mimic molecules were primarily known for use in cancer treatment. Accordingly, we sought to verify the therapeutic effects of the above active ingredients on obesity, dementia, and fibrosis as new applications. Therefore, active ingredients including antibodies that specifically bind to the LRR domain were prepared according to the methods described in existing patent literature and papers. Among them, active ingredients including antibodies that specifically bind to the LRR domain for which pharmacological effects were confirmed are shown in Table 5, and the numbers in Table 5 correspond to the numbers in FIGS. 17 to 19. Furthermore, to avoid unnecessary duplication, the specific manufacturing methods of the active ingredients have been omitted as they are described in detail in the patent literature listed in Table 5, and the entirety of the patent literature listed in Table 5 is cited by reference in the present invention.

[0178] No. Patent Document No. Active Ingredient Name (Clone Name) 3US 2024-0141036 A1GTC210-035 US 2025-0051444 A1L15-06C6 US 2024-0175880 A1LRRC4 Family Mimic Molecule 8US 2024-0132593 A1474.112EP 4056586 A1GTC110-0413US 2022-0249684 A1GTC110-0415US 2022-0213186 A1GTC210-0118US 2022-0275080 A1A721US 2021-0347849 A1L1-Fc23US 2021-0214424 A1LRRN1 E3624US 2021-0009710 A1anti-human LRIT2 antibody (Novusbio, NBP1-90876)25US 2021-0363244 A1H626US 11820801 B2L1-Fc29US 10870692 B2variable lymphocyte receptors (VLRs)32US 11365249 B2C835US 11136383 B2anti-LRRC33 antibody39US 10745487 B218G7H6A345EP 3262069 B115C449US 2016-0166684 A1131R01051US 9631024 B21C553EP 3120145 B1LRRC15 protein61EP 3421486 B1Anti-Mouse VISTA Antibody (13F3)66US 2012-0230979 A1Sp35-Fc69EP 3153521 B1Anti-Mouse VISTA Antibody (13F3)76US 8691222 B23D1.6.984US 2005-0271655 A114D587EP 2163561 A1the NOGO-receptor

[0179] 2. Confirmation of obesity treatment effect – Inhibition of lipid droplet formation during adipocyte differentiation

[0180] To evaluate the anti-obesity effect by inhibiting lipid droplet formation during adipocyte differentiation, 3T3-L1 preadipocytes or human adipose-derived stem cell / hMSC-derived adipocytes were used. First, cells were cultured to a confluent state, and adipogenesis was initiated using differentiation induction medium (usually containing IBMX, dexamethasone, insulin, and, if necessary, rosiglitazone). The test compounds were either administered simultaneously from the start of differentiation induction or divided among specific differentiation stages (early 0–2 days, mid-2 days, late 4–8 days). The differentiation period was typically about 6–10 days for 3T3-L1 cells, during which the compounds were repeatedly administered while the medium was replaced every two days. At the end of the experiment, intracellular triglyceride accumulation and lipid droplet formation were observed using Oil Red O staining or BODIPY / Nile Red fluorescent staining, and the number of droplets, droplet size, and lipid-positive area were quantified through microscopic image analysis.

[0181] As a result, as shown in Figure 17, it was confirmed that antibodies binding to the LRR domain can inhibit lipid droplet formation during lipid cell differentiation, thereby treating metabolic diseases including obesity.

[0182]

[0183] 3. Confirmation of Dementia Treatment Effect – Inhibition of Tau Protein Aggregation

[0184] To confirm the therapeutic effect of antibodies specifically binding to DKK-1 on dementia, tau protein aggregation and seeding assays were performed. Aggregation was induced in tau RD-P301S biosensor cells or full-length tau-expressing cells by treating them with preformed tau fibril or patient-derived tau seeds. Subsequently, whether the antibody specifically binding to DKK-1 provided in this invention inhibited aggregation was analyzed using Thioflavin S / T staining, filter trap assay, sarkosyl-insoluble tau fraction, and FRET-based aggregation assay. In immunohistochemistry, abnormal tau aggregate formation was quantified using AT8, MC1, and T22 oligomeric tau antibodies, and intracellular aggregate burden was measured using a confocal microscope.

[0185] As a result, as shown in Figure 18, it was confirmed that antibodies binding to the LRR domain can inhibit tau protein aggregation and treat neurological diseases including dementia.

[0186]

[0187] 4. Therapeutic effect on fibrosis

[0188] Human lung fibroblasts (MRC-5, HFL1, IMR-90, or primary human lung fibroblasts) were cultured in medium containing 10% FBS and 1% penicillin / streptomycin at 37°C under 5% CO2 conditions, and then seeded into 6-well or 12-well plates at appropriate densities until 70–80% confluence was reached. Subsequently, serum starvation was performed for 12–24 hours in low-serum medium containing 0.5–1% FBS to lower baseline signals and induce a uniform TGF-β1 response. To evaluate the antifibrotic effect of the test compounds, the compounds were pretreated to the cells for 1–2 hours or treated simultaneously with TGF-β1, and TGF-β1 was added to a final concentration of 2–10 ng / mL to induce myofibroblast conversion. The control group consisted of a vehicle-treated control, a TGF-β1-alone treatment group, a test compound-alone treatment group, and a TGF-β1+test compound treatment group. Generally, Smad2 / 3 phosphorylation was analyzed after 24 hours of treatment, and changes in the expression of fibrosis markers were evaluated after 48–72 hours of treatment. After the experiment, RNA and protein were recovered and subjected to qPCR and Western blot, respectively, and morphological changes were confirmed through immunocytochemistry in parallel.

[0189] As a result, as shown in Figure 19, it was confirmed that antibodies binding to the LRR domain can inhibit the conversion of fibroblasts into myofibroblasts, thereby treating fibrosis including liver and lung fibrosis.

[0190]

[0191] Although the present invention has been described in detail above, the scope of the present invention is not limited thereto, and it will be obvious to those skilled in the art that various modifications and variations are possible within the scope of the technical concept of the present invention as described in the claims.

Claims

A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising, as an active ingredient, a binding molecule that specifically binds to an LRR domain. In paragraph 1, The above-mentioned bonding molecule is, Heavy chain complementarity-determining region 1 (CDR-H1) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 1, 7, 14, 20, 26, 32, 38, 45, 51, 58, 64, 70, 76, 82, 88, and 96; Heavy chain complementarity-determining region 2 (CDR-H2) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 2, 8, 15, 21, 27, 33, 39, 46, 52, 59, 65, 71, 77, 83, 89, and 97; and A heavy chain variable region comprising: a heavy chain complementarity-determining region 3 (CDR-H3) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 3, 9, 16, 22, 28, 34, 40, 47, 53, 60, 66, 72, 78, 84, 90, and 98; and Light chain complementarity-determining region 1 (CDR-L1) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 4, 10, 17, 23, 29, 35, 41, 48, 54, 61, 67, 73, 79, 85, 91, and 99; Light chain complementarity-determining region 2 (CDR-L2) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 5, 11, 18, 24, 30, 36, 42, 49, 55, 62, 68, 74, 80, 86, 92, and 100; and A pharmaceutical composition comprising a binding molecule comprising: a light chain complementarity-determining region 3 (CDR-L3) comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs 6, 12, 19, 25, 31, 37, 43, 50, 56, 63, 69, 75, 81, 87, 93, and 101; and a light chain variable region comprising a light chain variable region. In paragraph 2, A pharmaceutical composition in which the above-mentioned binding molecule is selected from the following groups. (1) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 1; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 2; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 3; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 4; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 5; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 6; (2) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 7; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 8; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 9; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 10; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 11; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 12; (3) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 14; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 15; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 16; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 17; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 18; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 19; (4) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 20; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 21; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 22; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 23; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 24; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 25; (5) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 26; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 27; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 28; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 29; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 30; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 31; (6) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 32; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 33; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 34; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 35; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 36; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 37; (7) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 38; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 39; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 40; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 41; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 42; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 43; (8) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 45; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 46; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 47; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 48; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 49; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 50; (9) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 51; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 52; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 53; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 54; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 55; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 56; (10) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 58; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 59; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 60; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 61; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 62; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 63; (11) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 64; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 65; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 66; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 67; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 68; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 69; (12) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 70; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 71; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 72; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 73; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 74; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 75; (13) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 76; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 77; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 78; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 79; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 80; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 81; (14) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 82; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 83; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 84; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 85; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 86; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 87; (15) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 88; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 89; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 90; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 91; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 92; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 93; (16) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 96; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 97; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 98; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) comprising the amino acid sequence of SEQ ID NO. 99; a light chain complementarity-determining region 2 (CDR-L2) comprising the amino acid sequence of SEQ ID NO. 100; and a light chain complementarity-determining region 3 (CDR-L3) comprising the amino acid sequence of SEQ ID NO. 101; and (17) anti-human LRIT2 antibody (Novusbio, NBP1-90876). In any one of paragraphs 1 through 3, A pharmaceutical composition in which the above-mentioned binding molecule further comprises an Fc region (Fragment crystallization region) or a constant region. In paragraph 4, A pharmaceutical composition wherein the above Fc region is the Fc region of an IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, or a hybrid Fc region. In paragraph 1, A pharmaceutical composition in which the above-mentioned binding molecule is an antibody or a binding fragment thereof. In paragraph 6, A pharmaceutical composition wherein the antibody or its binding fragment is a chimeric antibody, humanized antibody, bivalent, amphoteric molecule, minibody, domain antibody, bispecific antibody, antibody mimic, unibody, diabody, triabody, tetrabody, or a fragment thereof. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising, as an active ingredient, a nucleic acid molecule encoding a binding molecule of any one of claims 1 to 3. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising, as an active ingredient, an expression vector into which a nucleic acid molecule encoding a binding molecule of any one of claims 1 to 3 has been inserted. (1) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 1; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 2; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 3; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 4; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 5; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 6; (2) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 7; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 8; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 9; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 10; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 11; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 12; (3) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 14; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 15; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 16; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 17; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 18; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 19; (4) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 20; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 21; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 22; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 23; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 24; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 25; (5) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 26; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 27; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 28; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 29; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 30; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 31; (6) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 32; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 33; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 34; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 35; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 36; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 37; (7) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 38; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 39; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 40; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 41; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 42; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 43; (8) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 45; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 46; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 47; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 48; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 49; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 50; (9) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 51; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 52; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 53; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 54; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 55; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 56; (10) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 58; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 59; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 60; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 61; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 62; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 63; (11) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 64; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 65; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 66; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 67; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 68; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 69; (12) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 70; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 71; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 72; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 73; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 74; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 75; (13) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 76; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 77; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 78; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 79; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 80; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 81; (14) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 82; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 83; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 84; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 85; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 86; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 87; (15) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 88; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 89; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 90; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 91; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 92; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 93; (16) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 96; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 97; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 98; and a heavy chain variable region comprising A binding molecule comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) comprising the amino acid sequence of SEQ ID NO. 99; a light chain complementarity-determining region 2 (CDR-L2) comprising the amino acid sequence of SEQ ID NO. 100; and a light chain complementarity-determining region 3 (CDR-L3) comprising the amino acid sequence of SEQ ID NO. 101; and (17) Anti-human LRIT2 antibody (Novusbio, NBP1-90876); and an antibody-drug conjugate (ADC) comprising a drug. In a chimeric antigen receptor (CAR) comprising an antigen-specific binding domain, a linkage domain, and a CD3 zeta (ζ) signaling domain, The above antigen-specific binding domain is a chimeric antigen receptor selected from the following group: (1) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 1; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 2; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 3; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 4; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 5; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 6; (2) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 7; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 8; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 9; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 10; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 11; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 12; (3) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 14; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 15; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 16; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 17; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 18; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 19; (4) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 20; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 21; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 22; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 23; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 24; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 25; (5) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 26; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 27; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 28; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 29; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 30; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 31; (6) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 32; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 33; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 34; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 35; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 36; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 37; (7) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 38; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 39; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 40; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 41; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 42; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 43; (8) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 45; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 46; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 47; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 48; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 49; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 50; (9) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 51; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 52; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 53; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 54; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 55; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 56; (10) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 58; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 59; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 60; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 61; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 62; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 63; (11) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 64; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 65; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 66; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 67; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 68; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 69; (12) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 70; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 71; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 72; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 73; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 74; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 75; (13) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 76; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 77; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 78; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 79; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 80; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 81; (14) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 82; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 83; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 84; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 85; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 86; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 87; (15) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 88; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 89; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 90; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) containing the amino acid sequence of SEQ ID NO. 91; a light chain complementarity-determining region 2 (CDR-L2) containing the amino acid sequence of SEQ ID NO. 92; and a light chain complementarity-determining region 3 (CDR-L3) containing the amino acid sequence of SEQ ID NO. 93; (16) a heavy chain complementarity-determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO. 96; a heavy chain complementarity-determining region 2 (CDR-H2) comprising the amino acid sequence of SEQ ID NO. 97; a heavy chain complementarity-determining region 3 (CDR-H3) comprising the amino acid sequence of SEQ ID NO. 98; and a heavy chain variable region comprising An antigen-specific binding domain comprising a heavy chain variable region including a light chain complementarity-determining region 1 (CDR-L1) comprising the amino acid sequence of SEQ ID NO. 99; a light chain complementarity-determining region 2 (CDR-L2) comprising the amino acid sequence of SEQ ID NO. 100; and a light chain complementarity-determining region 3 (CDR-L3) comprising the amino acid sequence of SEQ ID NO. 101; and (17) Antigen-specific binding domain containing anti-human LRIT2 (Novusbio, NBP1-90876). A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising an LRRC4 family mimic molecule of SEQ ID NO. 13 as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising the Lrig-1-Fc fusion protein of SEQ ID NO. 44 as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising variable lymphocyte receptors (VLRs) as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising LRRC15 protein as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases or fibrosis comprising an anti-mouse VISTA antibody (13F3) as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising the Sp35-Fc fusion protein of SEQ ID NO. 95 as an active ingredient. A pharmaceutical composition for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising the NOGO-receptor of SEQ ID NO. 102 as an active ingredient. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering a binding molecule that specifically binds to an LRR domain to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering an LRRC4 family mimic molecule of SEQ ID NO. 13 to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering the Lrig-1-Fc fusion protein of SEQ ID NO. 44 to a subject. A method for the prevention or treatment of metabolic diseases, neurological diseases, or fibrosis comprising the step of administering variable lymphocyte receptors (VLRs) to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering LRRC15 protein to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering an anti-mouse VISTA antibody (13F3) to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering the Sp35-Fc fusion protein of SEQ ID NO. 95 to a subject. A method for preventing or treating metabolic diseases, neurological diseases, or fibrosis, comprising the step of administering the NOGO-receptor of SEQ ID NO. 102 to a subject. In any one of paragraphs 12 through 18, A pharmaceutical composition in which the above fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin. In any one of paragraphs 12 through 18, A pharmaceutical composition wherein the above metabolic disease is one or more selected from the group consisting of insulin resistance disease, obesity, diabetes mellitus, dyslipidemia, liver disease, kidney damage, arteriosclerosis, and hypertension. In any one of paragraphs 12 through 18, A pharmaceutical composition in which the above-mentioned neurological disease is a neurodegenerative disease or a neuroinflammatory disease. In any one of paragraphs 12 through 18, A pharmaceutical composition wherein the above-mentioned neurodegenerative disease or neuroinflammatory disease is selected from the group consisting of stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Niemann-Pick disease, multiple sclerosis, prion disease, Creutzfeldt-Jakob disease, frontotemporal dementia, Lewy dementia, amyotrophic lateral sclerosis, paraneoplastic syndrome, corticobasal degeneration, multiple system atrophy, progressive supranuclear palsy, neurological autoimmune disease, spinocerebellar ataxia, inflammatory and neuropathic pain, cerebrovascular disease, spinal cord injury, and tauopathy.

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