Composition for preventing or treating brain and nervous system diseases

Abstract

The present invention relates to a composition for preventing, ameliorating, or treating cranial nervous system diseases using a binding molecule that can specifically bind to Lrig-1 protein, a protein present on the surface of regulatory T cells.

Description

[Technical field]

[0001] The present invention relates to a composition for preventing or treating a brain and nervous system disease. [Background technology]

[0002] In degenerative neurological diseases such as stroke, dementia, and Alzheimer's disease, declines in memory, attention, cognitive ability, and emotional regulation are observed, which are the result of neuronal death and atrophy of neural branches. The extension of neural branches of neural cells increases neuroplasticity and plays an important role in the memory and learning functions of neural circuits. Therefore, it is predicted that active ingredients that promote the extension of neural branches of neural cells and nerve regeneration may be developed as new therapeutic agents for degenerative neurological diseases.

[0003] With the rapid increase in the aging population, the incidence of degenerative nervous system diseases is also on the rise, and despite the innovative development of medicine, the prevention and treatment methods for degenerative nervous system diseases are still unclear, and no definitively effective drugs have been found. Currently, drugs and treatment methods for degenerative nervous system diseases are being developed, but in many cases they cause side effects and toxicity due to long-term use, and they only have the effect of alleviating symptoms rather than treating them, so there is an urgent need to develop materials that can reduce side effects and toxicity and treat them. Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a composition for various applications capable of preventing, ameliorating or treating cranial nervous system diseases, particularly neurodegenerative diseases or neuroinflammatory diseases.

[0005] However, the technical problem that the present invention aims to achieve is not limited to the problem described above, and other problems not described will be clearly understood by those having ordinary skill in the art from the following description. [Means for solving the problem]

[0006] According to one embodiment of the present invention, the present invention relates to a composition for preventing, ameliorating, or treating a brain and nervous system disease, comprising as an active ingredient a binding molecule that specifically binds to Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present in regulatory T cells (Treg cells).

[0007] As used herein, a "binding molecule" refers to an intact immunoglobulin, including a monoclonal antibody, such as a chimeric, humanized or human monoclonal antibody, or an immunoglobulin that binds an antigen, e.g., a variable domain containing immunoglobulin fragment that competes with the intact immunoglobulin for binding to monomeric or trimeric HA of influenza A virus. Regardless of structure, the antigen-binding fragment binds to the same antigen recognized by the intact immunoglobulin. Antigen-binding fragments can include peptides or polypeptides that include an amino acid sequence of two or more contiguous groups of the amino acid sequence of the binding molecule, 20 or more contiguous amino acid residues, 25 or more contiguous amino acid residues, 30 or more contiguous amino acid residues, 35 or more contiguous amino acid residues, 40 or more contiguous amino acid residues, 50 or more contiguous amino acid residues, 60 or more contiguous amino acid residues, 70 or more contiguous amino acid residues, 80 or more contiguous amino acid residues, 90 or more contiguous amino acid residues, 100 or more contiguous amino acid residues, 125 or more contiguous amino acid residues, 150 or more contiguous amino acid residues, 175 or more contiguous amino acid residues, 200 or more contiguous amino acid residues, or 250 or more contiguous amino acid residues. "Antigen-binding fragments" include, inter alia, Fab, F(ab'), F(ab')2, Fv, dAb, Fd, complementarity determining region (CDR) fragments, single chain antibodies (scFv), bivalent single chain antibodies, single chain phage antibodies, diabodies, triabodies, tetrabodies, polypeptides containing one or more fragments of an immunoglobulin sufficient to bind a specific antigen to a polypeptide. The fragments can be produced synthetically or by enzymatic or chemical degradation of intact immunoglobulins, or genetically engineered by recombinant DNA technology. Methods of production are well known in the art.

[0008] In the present invention, the "Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein" is a transmembrane protein consisting of 1091 amino acids present on the surface of regulatory T cells, and is composed of an extracellular or lumenal leucine repeat (LRR), three immunoglobulin-like domains, a transmembrane sequence, and a cytoplasmic tail. The LRIG gene family includes LRIG1, LRIG2, and LRIG3, and the amino acids between them are highly conserved. The LRIG1 gene is highly expressed in normal skin, and is expressed in basal and hair follicle cells to regulate the proliferation of epithelial stem cells. Therefore, it plays an important role in maintaining epidermal homeostasis, and its absence may lead to psoriasis or skin cancer. It has been reported that when the chromosome 3p14.3 region where LRIG1 is located is broken, it may develop into cancer cells, and in fact, it has been confirmed that the expression of LRIG1 is significantly reduced in renal cell carcinoma and cutaneous squamous cell carcinoma. However, it has been recently determined that only about 20 to 30% of cancers express Lrig-1. Meanwhile, 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.

[0009] In the present invention, the Lrig-1 protein may be a polypeptide represented by SEQ ID NO: 1, which is derived from humans, or a polypeptide represented by SEQ ID NO: 3, which is derived from mice, but is not limited thereto.

[0010] In addition, in the present invention, the Lrig-1 protein represented by SEQ ID NO: 1 may be encoded by a polynucleotide represented by SEQ ID NO: 2, but is not limited thereto.

[0011] Furthermore, in the present invention, the Lrig-1 protein represented by SEQ ID NO: 3 may be encoded by a polynucleotide represented by SEQ ID NO: 4, but is not limited thereto.

[0012] In the present invention, the binding molecule is A heavy chain variable region comprising: a heavy chain CDR1 consisting of the amino acid sequence represented by SEQ ID NO:5 or 13; a heavy chain CDR2 consisting of the amino acid sequence represented by SEQ ID NO:6 or 14; and a heavy chain CDR3 consisting of the amino acid sequence represented by SEQ ID NO:7 or 15; and The binding molecule may comprise a light chain variable region including: a light chain CDR1 represented by the amino acid sequence represented by SEQ ID NO: 8 or 16; a light chain CDR2 represented by the amino acid sequence represented by SEQ ID NO: 9 or 17; and a light chain CDR3 represented by the amino acid sequence represented by SEQ ID NO: 10 or 18.

[0013] In the present invention, the binding molecule is (a) a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO:5, a heavy chain CDR2 represented by SEQ ID NO:6, and a heavy chain CDR3 represented by SEQ ID NO:7; and (b) a heavy chain variable region selected from the group consisting of a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO: 13, a heavy chain CDR2 represented by SEQ ID NO: 14, and a heavy chain CDR3 represented by SEQ ID NO: 15; and (c) a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO:8, a light chain CDR2 represented by SEQ ID NO:9, and a light chain CDR3 represented by SEQ ID NO:10; and (d) a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO: 16, a light chain CDR2 represented by SEQ ID NO: 17, and a light chain CDR3 represented by SEQ ID NO: 18;

[0014] In the present invention, the binding molecule is (1) a binding molecule comprising a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO:5, a heavy chain CDR2 represented by SEQ ID NO:6, and a heavy chain CDR3 represented by SEQ ID NO:7; and a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO:8, a light chain CDR2 represented by SEQ ID NO:9, and a light chain CDR3 represented by SEQ ID NO:10; and (2) A binding molecule selected from the group consisting of a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO: 13, a heavy chain CDR2 represented by SEQ ID NO: 14, and a heavy chain CDR3 represented by SEQ ID NO: 15; and a binding molecule comprising a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO: 16, a light chain CDR2 represented by SEQ ID NO: 17, and a light chain CDR3 represented by SEQ ID NO: 18.

[0015] In the present invention, the binding molecule is A heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or 19; and The binding molecule may comprise a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or 20.

[0016] In the present invention, the binding molecule is A binding molecule comprising a heavy chain variable region represented by SEQ ID NO:11 and a light chain variable region represented by SEQ ID NO:12; and a binding molecule comprising a heavy chain variable region represented by SEQ ID NO:19 and a light chain variable region represented by SEQ ID NO:20.

[0017] In the present invention, the binding molecule may further comprise an Fc region (fragment crystallization region) or a constant region, which may be or be derived from an Fc region of an IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3 or IgG4 antibody, or may be a hybrid Fc region.

[0018] In the present invention, the Fc region may be an Fc region of a mammalian-derived IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, and preferably an Fc region of a human-derived IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3, or IgG4 antibody, but is not limited thereto.

[0019] As an example of the present invention, the constant region may be a mouse-derived IgG2a constant region represented by SEQ ID NO: 21, but is not limited thereto.

[0020] As an example of the present invention, the constant region may be, but is not limited to, a mouse-derived immunoglobulin kappa constant region represented by SEQ ID NO: 22.

[0021] As an example of the present invention, the constant region may be a human-derived IgG1 constant region represented by SEQ ID NO: 23 or 24, but is not limited thereto.

[0022] As an example of the present invention, the constant region may be a human-derived immunoglobulin kappa constant region represented by SEQ ID NO: 25, but is not limited thereto.

[0023] As an example of the present invention, the constant region may be the human-derived IgG2 constant region represented by SEQ ID NO: 26, but is not limited thereto.

[0024] As an example of the present invention, the constant region may be a human-derived IgG3 constant region represented by SEQ ID NO: 27, but is not limited thereto.

[0025] As an example of the present invention, the constant region may be a human-derived IgG4 constant region represented by SEQ ID NO: 28, but is not limited thereto.

[0026] As an example of the present invention, the Fc region may be, but is not limited to, a human-derived immunoglobulin lambda constant region.

[0027] In the present invention, the "hybrid Fc" is derived from a combination of human IgG subclasses or a combination of human IgD and IgG. When the hybrid Fc is bound to a biologically active molecule, such as a polypeptide, 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.

[0028] As an example of the present invention, the hybrid Fc region may be a hybrid Fc region represented by SEQ ID NO:29, but is not limited thereto.

[0029] In the binding molecule of the present invention, the Fc or constant region can be linked to the variable region via a linker, in which the linker may be linked to the C-terminus of the Fc or constant region, and the N-terminus of the binding molecule of the present invention may be linked to the linker, but is not limited thereto.

[0030] In the present invention, the "linker" may include a sequence that can be cleaved by an enzyme that is overexpressed in tissues or cells of a target disease. When the linker is cleavable by an enzyme that is overexpressed as described above, it is possible to effectively prevent the activity of the polypeptide from being reduced by the Fc or invariant region. In the present invention, a preferred example of the linker may be a peptide linker consisting of 33 amino acids located at positions 282 to 314 of human albumin, which is the most abundant in blood, and more preferably a peptide linker consisting of 13 amino acids located at positions 292 to 304, and these portions are mostly exposed to the outside in a three-dimensional structure, and are therefore the portions that are least likely to induce an immune response in the body. However, the present invention is not limited thereto.

[0031] A binding molecule of the invention can further comprise a heavy chain constant region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs:21, 23, 24, 26, 27, 28 and 29.

[0032] A binding molecule of the invention can further comprise a light chain constant region consisting of the amino acid sequence set forth in SEQ ID NO:22 or 25.

[0033] The binding molecule of the invention comprises A heavy chain constant region consisting of the amino acid sequence represented by SEQ ID NO: 21; and It may further comprise a light chain constant region consisting of the amino acid sequence shown in SEQ ID NO:22.

[0034] The binding molecule of the invention comprises A heavy chain constant region consisting of the amino acid sequence represented by SEQ ID NO: 23, 24, 26, 27 or 28; and It may further comprise a light chain constant region consisting of the amino acid sequence shown in SEQ ID NO:25.

[0035] The binding molecule of the invention comprises It may further comprise a heavy chain constant region consisting of the amino acid sequence shown in SEQ ID NO:29.

[0036] The binding molecule of the invention comprises a binding molecule comprising a heavy chain represented by SEQ ID NO: 30 and a light chain represented by SEQ ID NO: 31; A binding molecule comprising a heavy chain represented by SEQ ID NO: 32 and a light chain represented by SEQ ID NO: 33; and a binding molecule comprising a heavy chain represented by SEQ ID NO:34, and a light chain represented by SEQ ID NO:35.

[0037] The binding molecule of the present invention is characterized in that it is an antibody, but is not limited thereto. The antibody may be a monoclonal antibody, a full-length antibody, or a portion of an antibody, and includes all antibody fragments that have the ability to bind to the Lrig-1 protein and can bind to the Lrig-1 antigenic determinant site competitively with the binding molecule of the present invention.

[0038] In the present invention, the "antibody" refers to a protein molecule that plays the role of a receptor that specifically recognizes an antigen, including 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 regulatory T cells. Preferably, the antibody may specifically recognize the leucine rich region or immunoglobulin-like domain of the Lrig-1 protein, but is not limited thereto.

[0039] In the present invention, the "immunoglobulin" has a heavy chain and a light chain, each of which contains a constant region and a variable region. The variable regions of the light chain and the heavy chain contain three variable regions called complementarity determining regions (hereinafter referred to as "CDRs") and four framework regions. The CDRs mainly play a role in binding to an epitope of an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2 and CDR3, sequentially starting from the N-terminus, and are also identified by the chain in which the particular CDR is located.

[0040] In the present invention, the term "monoclonal antibody" refers to an antibody molecule of a single molecular composition obtained from a population of substantially identical antibodies, and exhibits a single binding specificity and affinity for a specific antigenic determinant (epitope).

[0041] In the present invention, the "full-length antibody" has a structure having two full-length light chains and two full-length heavy chains, each of which is linked to a heavy chain by a disulfide bond, and includes IgA, IgD, IgE, IgM, and IgG. The IgG subtypes include IgG1, IgG2, IgG3, and IgG4.

[0042] In the present invention, the "antibody fragment" refers to a fragment having an antigen-binding function, and includes Fab, Fab', F(ab')2, and Fv. The Fab has a structure including a light chain and a heavy chain variable region, a light chain constant region, and the first constant region (CH1 domain) of the heavy chain, and has one antigen-binding site. Furthermore, Fab' is different 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 by disulfide bonds between the cysteine ​​residues in the hinge region of Fab'. Fv (variable fragment) refers to the smallest antibody fragment having only a heavy chain variable region and a light chain variable region. In a double chain Fv (dsFv), the heavy chain variable region and the light chain variable region are linked by a disulfide bond, and in a single chain Fv (scFv), the heavy chain variable region and the light chain variable region are generally linked by a covalent bond via a peptide linker. The antibody fragment can be produced by recombinant gene technology, such as when proteolytic enzymes, such as papain or pepsin, are used to obtain Fab or F(ab')2 fragments.

[0043] In addition, in the present invention, the antibody may be, but is not limited to, 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.

[0044] In the present invention, the "chimeric antibody" is an antibody obtained by recombining the variable region of a mouse antibody and the constant region of a human antibody, and has a significantly improved immune response compared to mouse antibodies.

[0045] 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 has been modified to resemble an antibody mutant naturally produced by humans. For example, the humanized antibody can be produced by recombining mouse-derived CDRs with human antibody-derived FRs to produce a humanized variable region, which can then be recombined with a preferred human antibody constant region to produce a humanized antibody.

[0046] In the present invention, the binding molecule can also be provided as a bispecific antibody or bispecific antigen-binding fragment capable of binding to the Lrig-1 protein and also to other proteins.

[0047] In the present invention, the bispecific antibodies and bispecific antigen-binding fragments may comprise a binding molecule according to the present invention. In one example, the bispecific antibodies and bispecific antigen-binding fragments may comprise an antigen-binding domain capable of binding to the Lrig-1 protein, where the antigen-binding domain capable of binding to the Lrig-1 protein may comprise or consist of a binding molecule according to the present invention.

[0048] The bispecific antibodies and bispecific antigen-binding fragments provided herein comprise an antigen-binding domain that is a binding molecule capable of binding to the Lrig-1 protein according to the present invention, and an antigen-binding domain that is capable of binding to another target protein, where the antigen-binding domain that is capable of binding to another target protein may be, but is not limited to, an antigen-binding domain that is capable of binding to a protein other than the Lrig-1 protein, for example, but is not limited to, PD-1 or a cell surface receptor.

[0049] The bispecific antibodies and bispecific antigen-binding fragments according to the invention can be provided in any suitable format, for example as described in the documents fully incorporated herein by reference (see Kontermann MAbs 2012, 4(2):182-197). For example, the bispecific antibodies or bispecific antigen-binding fragments can be provided as 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 2in1-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 pair or SEED-body), small bispecific antibody molecules (e.g. diabodies (Db), dsDb, DART, scDb, tandAbs, tandem scFv (taFv), tandem dAb / VHH, triple body, triple head, Fab-scFv, or F(ab')2-scFv2), bispecific Fc and CH3 fusion proteins (e.g. taFv-Fc, di-diabody, scDb-CH3, scFv-Fc-scFv, HCAb-VHH, scFv-kih-Fc, or scFv-kih-CH3), or bispecific fusion proteins (e.g. scFv2-albumin, scDb-albumin, taFv-toxin, DNL-Fab3, DNL-Fab4-IgG, DNL-Fab4-IgG-cytokine2). See in particular Figure 2 of the document (ref: Kontermann MAbs 2012, 4(2):182-19). The skilled person is capable of designing and producing bispecific antibodies and bispecific antigen-binding fragments according to the invention.

[0050] In the present invention, methods for producing the bispecific antibodies include chemical cross-linking of antibodies or antibody fragments with reducible disulfide or non-reducible thioether bonds, as described, for example, in Segal and Bast, 2001. Production of Bispecific Antibodies. Current Protocols in Immunology. 14:IV:2.13:2.13.1-2.13.16, the entire contents of which are incorporated herein by reference. For example, N-succinimidyl-3-(-2-pyridyldithio)-propionate (SPDP) can be used to chemically cross-link Fab fragments, for example, via hinge region SH-groups, to generate disulfide-linked bispecific F(ab)2 heterodimers.

[0051] Also in accordance with the present invention, another method for producing said bispecific antibodies comprises fusing antibody-producing hybridomas, for example with polyethylene glycol, to generate quadroma cells capable of secreting bispecific antibodies, as described, for example, in the literature (see: DM and Bast, BJ 2001. Production of Bispecific Antibodies. Current Protocols in Immunology. 14:IV:2.13:2.13.1-2.13.16).

[0052] The bispecific antibodies and bispecific antigen-binding fragments according to the invention can further be recombinantly produced by expression from a nucleic acid construct encoding the polypeptide for the antigen-binding molecule, e.g. as described in Antibody Engineering: Methods and Protocols, Second Edition (Humana Press, 2012), at Chapter 40: Production of Bispecific Antibodies: Diabodies and Tandem scFv (Hornig and Farber-Schwarz), or French, How to make bispecific antibodies, Methods Mol. Med. 2000; 40: 333-339, both of which are incorporated herein by reference in their entirety.

[0053] For example, a DNA construct encoding light and heavy chain variable domains for two antigen binding domains (i.e., light and heavy chain variable domains for an antigen binding domain capable of binding, e.g., PD-1, and light and heavy chain variable domains for an antigen binding domain capable of binding another target protein), and containing sequences encoding suitable linkers or dimerization domains between the antigen binding domains, can be produced by molecular cloning techniques. Recombinant bispecific antibodies can then be produced by expression (e.g., in vitro) of the construct in a suitable host cell (e.g., a mammalian host cell), and the expressed recombinant bispecific antibodies can then be optionally purified.

[0054] Antibodies can be produced by an affinity maturation process, which produces modified antibodies that have improved affinity for the antigen compared to the unmodified parent antibody. Affinity matured antibodies can be produced by procedures known in the art, e.g., in the literature (see: Marks et al., Rio / Technology 10:779-783 (1992); Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-159 (1995); and Hawkins et al, J. Mol. Biol. 226:889-896 (1992)).

[0055] At the same time, the binding molecule provided by the present invention can include variants of the amino acid sequence, as long as it can specifically bind to Lrig-1 protein. For example, the amino acid sequence of the antibody can be altered to improve the binding affinity and / or other biological properties of the antibody. Such modifications include, for example, deletion, insertion and / or substitution of residues in the amino acid sequence of the antibody.

[0056] Such amino acid mutations are made based on the relative similarity of the amino acid side chain substitutes, e.g., hydrophobicity, hydrophilicity, charge, size, etc. Analysis of the size, shape, and type of amino acid side chain substitutes 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. Thus, based on these considerations, arginine, lysine, and histidine; alanine, glycine, and serine; and phenylalanine, tryptophan, and tyrosine are biologically functional equivalents.

[0057] When making mutations, the hydropathic index of amino acids is considered. Each amino acid is assigned a hydrophobic 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 hydropathic amino acid index is very important in conferring interactive biological functions of proteins. It is a well-known fact that only by substituting amino acids with similar hydrophobic indexes can similar biological activities be retained. When introducing mutations with reference to the hydrophobic index, substitutions are made between amino acids showing a difference in hydrophobic index within preferably ±2, more preferably ±1, and even more preferably ±0.5.

[0058] On the other hand, it is also well known that substitutions between amino acids with similar hydrophilicity values ​​result in proteins with equivalent biological activity. As disclosed in U.S. Patent No. 4,554,101, the following hydrophilicity values ​​are assigned to each amino acid residue: arginine (+3.0); lysine (+3.0); aspartate (+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 mutations with reference to the hydrophilicity value, substitutions can be made between amino acids showing a difference in hydrophilicity value within preferably ±2, more preferably within ±1, and even more preferably within ±0.5.

[0059] Amino acid exchanges in proteins that do not globally alter the activity of the molecule are known in the art (H. Neurath, R.L. Hill, The Proteins, Academic Press, New York (1979)). The most commonly occurring exchanges are between the 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, Gln / Glu.

[0060] In view of the above mentioned biologically equivalent variants, the binding molecules of the present invention are also intended to include sequences that show substantial identity to the sequences set out in the sequence listing.

[0061] In the present invention, the term "substantial identity" refers to a sequence that shows at least 61% homology, more preferably 70% homology, even more preferably 80% homology, and most preferably 90% homology, when the sequence of the present invention is aligned with any other sequence for maximum correspondence and the aligned sequences are analyzed using algorithms commonly used in the art. Methods of alignment for comparing sequences are known in the art. Various methods and algorithms for alignment are disclosed in Smith and Waterman, Adv. Appl. Math. 2:482 (1981); Needleman and Wunsch, J. Mol. Bio. 48:443 (1970); Pearson and Lipman, Methods in Mol. Biol. 24:307-31 (1988); Higgins and Sharp, Gene 73:237-44 (1988); Higgins and Sharp, CABIOS 5:151-3 (1989); Corpet et al., Nuc. Acids Res. 16:10881-90 (1988); Huang et al., Comp. Appl. BioSci. 8:155-65 (1992) and Pearson et al., Meth. Mol. Biol. 24:307-31 (1994). The NCBI Basic Local Alignment Search Tool (BLAST) (Altschul et al., J. Mol. Biol. 215:403-10 (1990)) is available at the National Center for Biological Information (NBCI) and can be used in conjunction with sequence analysis programs such as blastp, blasm, blastx, tblastn and tblastx on the Internet. BLSAT can be accessed at http: / / www.ncbi.nlm.nih.gov / BLAST / . Instructions on how to compare sequence homology using this program can be found online at http: / / www.ncbi.nlm.nih.gov / BLAST / blast_help.html.

[0062] In the present invention, the binding molecule, preferably the antibody, can be produced by conventional methods for producing antibodies, but can also be produced by affinity maturation.

[0063] In the present invention, the term "affinity maturation" refers to a process in which activated B cells produce antibodies with increased affinity to an antigen during an immune response. For the purposes of the present invention, affinity maturation can be based on the principles of mutation and selection, as in the natural process, to produce antibodies or antibody fragments produced by affinity maturation.

[0064] The binding molecules, preferably antibodies, provided by the present invention can effectively prevent, ameliorate or treat brain and nervous system diseases, in particular neurodegenerative diseases or neuroinflammatory diseases.

[0065] According to another embodiment of the present invention, there is provided a composition for preventing, ameliorating or treating a brain and nervous system disease, comprising as an active ingredient a nucleic acid molecule encoding the binding molecule provided by the present invention; an expression vector into which the nucleic acid molecule has been inserted; or a host cell line transfected with the expression vector.

[0066] The nucleic acid molecules of the present invention include all nucleic acid molecules in which the amino acid sequence of the binding molecule provided by the present invention has been translated into a polynucleotide sequence as known to those skilled in the art. Therefore, various polynucleotide sequences can be produced by ORF (open reading frame), and all of these are included in the nucleic acid molecules of the present invention.

[0067] In the present invention, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double-stranded DNA to which additional DNA segments can be ligated. Another type of vector is a phage vector. Yet another type of vector is a viral vector, to which additional DNA segments can be ligated into the viral genome. Some vectors are capable of autonomous replication in the host cell into which they are introduced (e.g., bacterial vectors are episomal mammalian vectors with a bacterial origin of replication). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of the host cell upon introduction into the host cell, and are thereby replicated along with the host genome. Moreover, some vectors are capable of directing the expression of genes to which they are linked at an operational level. Such vectors are referred to herein as "recombinant expression vectors" or simply "expression vectors". In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector.

[0068] In the present invention, specific examples of the expression vector may be selected from the group consisting of pCDNA vectors, F, R1, RP1, Col, pBR322, ToL, Ti vectors, which are widely used commercially; 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. All expression vectors known to those skilled in the art as expression vectors can be used in the present invention, and the selection of an expression vector depends on the properties of the intended host cell. When a vector is introduced into a host cell, it is carried out by calcium phosphate transfection, virus infection, DEAE-dextran regulated transfection, lipofectamine transfection, or electroporation, but is not limited thereto, and those skilled in the art can select and use an introduction method suitable for the expression vector and host cell used. Preferably, the vector contains one or more selection markers, but is not limited thereto, and a vector without a selection marker can be used to select based on the presence or absence of product production. The selection of the selectable marker is determined by the intended host cell, and this is done using methods already known to those skilled in the art, and therefore the present invention is not limited thereto.

[0069] The nucleic acid molecule of the present invention can be fused to an expression vector by inserting a tag sequence to facilitate purification. The tag includes, but is not limited to, a hexa-histidine tag, a hemagglutinin tag, a myc tag, or a flag tag, and any tag known to those skilled in the art to facilitate purification can be used in the present invention.

[0070] In the present invention, the "host cell" includes an individual cell or cell culture that can be or has been a recipient of a vector for incorporation of a polypeptide insert. A host cell includes the progeny of a single host cell, which may not necessarily be completely identical (in morphology or genomic DNA complement) to the original parent cell due to natural, accidental or deliberate mutation. A host cell includes a cell transfected in vivo with a polypeptide of the present application.

[0071] In the present invention, the host cell may include cells of mammalian, plant, insect, fungal or cellular origin, for example, bacterial cells such as Escherichia coli, Streptomyces, Salmonella typhimurium, etc.; fungal cells such as yeast cells and Pichia pastoris, etc.; insect cells such as Drosophila and Spodoptera Sf9 cells, etc.; animal cells such as CHO (Chinese hamster ovary cells), SP2 / 0 (mouse myeloma), human lymphoblastoid, COS, NSO (mouse myeloma), 293T, Bowes melanoma cells, HT-1080, BHK (Baby Hamster Kidney cells), HEK (Human Embryonic Kidney cells), or PERC.6 (human retinal cells); or plant cells, but are not limited thereto, and any cells that can be used as a host cell line known to those skilled in the art can be used.

[0072] According to yet another embodiment of the present invention, there is provided a composition for preventing, ameliorating, or treating a brain and nervous system disease, comprising as an active ingredient an antibody-drug conjugate (ADC) comprising an antibody and a drug provided by the present invention.

[0073] In the present invention, the "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 the α-amine group at the N-terminus of the heavy chain and / or light chain of an antibody.

[0074] In the present invention, the "drug" may mean any substance having a specific biological activity in cells, including DNA, RNA, or peptide. The drug may be in a form containing a reactive group capable of reacting with an α-amine group to crosslink, or may be in a form linked to a linker containing a reactive group capable of reacting with an α-amine group to crosslink.

[0075] In the present invention, examples of reactive groups capable of reacting with the α-amine group to crosslink include, but are not limited to, all types known in the art that react with amine groups, 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. Examples of reactive groups include, but are not limited to, any one of isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonyl chlorides, aldehydes, glyoxals, epoxides, oxiranes, carbonates, aryl halides, imidoesters, carbodiimides, anhydrides, and fluorophenyl esters.

[0076] In the present invention, the above-mentioned drug is included regardless of the type, so long as it is a drug capable of treating a cranial nervous system disease, particularly a neurodegenerative disease or a neuroinflammatory disease.

[0077] The cranial nervous system disease to be prevented, ameliorated or treated by the composition provided by the present invention may be a neurodegenerative disease or a neuroinflammatory disease.

[0078] In the present invention, the "neurodegenerative disease" may mean a disease caused by a decrease or loss of function of nerve cells, and the "neuroinflammatory disease" may mean a disease caused by an excessive inflammatory response of the nervous system. In the present invention, specific examples of the neurodegenerative disease or neuroinflammatory disease include stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Niemann-Pick disease, multiple sclerosis, prion disease, Creutzfeldt-Jakob disease, frontotemporal dementia, dementia with Lewy bodies, amyotrophic lateral sclerosis, paraneoplastic syndrome, corticobasal degeneration, multiple system atrophy, progressive supranuclear palsy, autoimmune diseases of the nervous system, spinocerebellar ataxia, inflammatory and neuropathic pain, cerebrovascular disease, spinal cord injury, and the like. The present invention may be selected from the group consisting of, but not limited to, tauopathy, pulmonary arterial disease, and tauopathy.

[0079] Furthermore, the composition provided by the present invention can be used as a pharmaceutical composition or a food composition, but is not limited thereto.

[0080] The "prevention" of the present invention includes, without limitation, all actions that can block, suppress or delay symptoms caused by a neurodegenerative disease or a neuroinflammatory disease using the composition of the present invention.

[0081] The "treatment" and "amelioration" of the present invention include, without limitation, any action that can improve or benefit symptoms caused by a neurodegenerative disease or a neuroinflammatory disease using the composition of the present invention.

[0082] In the present invention, the pharmaceutical composition is characterized in that it is in the form of a capsule, tablet, granule, injection, ointment, powder or drink, and the pharmaceutical composition may be characterized in that it is intended for humans.

[0083] The pharmaceutical composition of the present invention can be formulated into oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, external preparations, suppositories, and sterile injection solutions by a conventional method, but is not limited thereto. The pharmaceutical composition of the present invention can contain a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a dye, a flavoring, etc., when administered orally, and may be mixed with a buffer, a preservative, a soothing agent, a solubilizer, an isotonic agent, a stabilizer, etc., when administered locally, and may be a base, an excipient, a lubricant, a preservative, etc., when administered locally. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with the above-mentioned pharmaceutically acceptable carrier. For example, for oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injection, it can be prepared in unit dose ampoules or multiple dose forms. In addition, it can be formulated into solutions, suspensions, tablets, capsules, sustained release preparations, etc.

[0084] On the other hand, 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, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil, etc. In addition, fillers, anti-agglomerating agents, lubricants, wetting agents, flavors, emulsifiers, preservatives, etc. may be additionally included.

[0085] The routes of administration of the pharmaceutical compositions according to the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal. Oral or parenteral administration is preferred.

[0086] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention can also be administered in the form of suppositories for rectal administration.

[0087] The pharmaceutical composition of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, formulation, administration time, administration route, excretion rate, drug composition, and the severity of the specific disease to be prevented or treated. The dosage of the pharmaceutical composition varies depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, and can be appropriately selected by those skilled in the art, and can be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be once or several times a day. The dosage does not limit the scope of the present invention in any aspect. The pharmaceutical composition of the present invention may be formulated into pills, sugar-coated tablets, capsules, liquids, gels, syrups, slurries, and suspensions.

[0088] Food compositions containing the composition of the present invention as an active ingredient can be manufactured into various food forms, such as beverages, gums, teas, vitamin complexes, powders, granules, tablets, capsules, sweets, rice cakes, bread, etc. The food compositions of the present invention are made of plant extracts that have almost no toxicity or side effects, and therefore can be safely used for long-term preventive use.

[0089] When the composition of the present invention is included in a food composition, the amount added may be in the range of 0.1 to 50% by weight of the total composition.

[0090] Here, when the food composition is manufactured in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and various flavorings or natural carbohydrates, etc., can be contained as additional ingredients, like ordinary beverages. That is, natural carbohydrates can include monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, ordinary sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. Examples of the flavorings include natural flavorings (thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavorings (saccharin, aspartame, etc.).

[0091] In addition, the food composition of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavorings such as synthetic flavors and natural flavors, colorants, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like.

[0092] Such components can be used independently or in combination. The proportion of such additives is not critical, but is generally selected from the range of 0.1 to about 50 parts by weight per 100 parts by weight of the composition of the present invention.

[0093] According to yet another embodiment of the present invention, there is provided a method for preventing, ameliorating or treating a brain and nervous system disease, comprising the step of administering to an individual in need thereof a binding molecule provided by the present invention; a nucleic acid molecule encoding the binding molecule; an expression vector into which the nucleic acid molecule has been inserted; a host cell line transfected with the expression vector; or an antibody-drug conjugate (ADC) provided by the present invention.

[0094] In the present invention, the "individual" refers to an individual suspected of developing a cranial nervous system disease, and the individual suspected of developing the disease refers to mammals including rodents, including humans, that have developed or may develop the disease, and livestock, but includes, without limitation, individuals that can be treated with the effective substance provided by the present invention.

[0095] The cranial nervous system disease to be treated by the method of the present invention may be a neurodegenerative disease or a neuroinflammatory disease, and specific examples thereof include stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Niemann-Pick disease, multiple sclerosis, prion disease, Creutzfeldt-Jakob disease, frontotemporal dementia, dementia with Lewy bodies, amyotrophic lateral sclerosis, paraneoplastic syndrome, corticobasal degeneration, multiple system atrophy, progressive supranuclear palsy, autoimmune diseases of the nervous system, spinocerebellar ataxia, inflammatory and neuropathic pain, cerebrovascular disease, spinal cord injury, and the like. The present invention may be selected from the group consisting of, but not limited to, tauopathy, pulmonary arterial disease, and tauopathy.

[0096] The methods of the invention may comprise administering a pharma- ceutical effective amount of a binding molecule provided by the invention; a nucleic acid molecule encoding the binding molecule; an expression vector into which the nucleic acid molecule has been inserted; a host cell line transfected with the expression vector; or an antibody-drug conjugate (ADC) provided by the invention.

[0097] The total amount of the suitable daily use can be determined by the treating physician within the scope of sound medical judgment, and can be administered in one or several doses. However, for the purposes of the present invention, the specific therapeutically effective amount for a particular patient is preferably applied differently depending on various factors including the type and degree of response to be achieved, the specific composition containing the active ingredient, including whether other preparations are used in some cases, the age, weight, general health condition, sex and diet of the patient, the administration time, administration route and excretion rate of the composition containing the active ingredient, the duration of treatment, drugs used with or simultaneously with the specific composition, and similar factors well known in the pharmaceutical art.

[0098] On the other hand, the method for preventing or treating a central nervous system disease may be, but is not limited to, a combination therapy that further includes administering a compound or substance having therapeutic activity against one or more diseases.

[0099] In the present invention, the term "combination" should be understood to indicate simultaneous, separate or sequential administration. If the administration is sequential or separate, the interval between administrations of the second components should be such that the beneficial effect of the combination is not lost.

[0100] In the present invention, the administration dose of the binding molecule or antibody-drug conjugate may be about 0.0001 μg to 500 mg per kg of patient body weight, but is not limited thereto.

[0101] According to yet another embodiment of the present invention, there is provided a composition for diagnosing a brain and nervous system disease, comprising the binding molecule provided by the present invention.

[0102] In the present invention, the term "diagnosis" means to confirm the presence or characteristics of a pathological condition, and for the purposes of the present invention, diagnosis means to confirm the presence or absence of onset of a cranial nervous system disease, particularly a neurodegenerative disease or a neuroinflammatory disease.

[0103] In the present invention, the expression level of Lrig-1 protein can be measured using the binding molecule, thereby making it possible to diagnose brain and nervous system diseases.

[0104] According to yet another embodiment of the present invention, there is provided a diagnostic kit for cranial nervous system diseases, comprising the diagnostic composition of the present invention.

[0105] In the present invention, the "kit" refers to a set of compositions and accessories required for a specific purpose. For the purposes of the present invention, the kit of the present invention can diagnose the disease by confirming the expression level of the Lrig-1 protein. The kit of the present invention includes a primer, a probe, or an antibody that selectively recognizes the marker for measuring the expression level of a diagnostic marker for an immune-related disease, as well as one or more other component compositions, solutions, or devices suitable for the analysis method.

[0106] In the present invention, the kit may be, but is not limited to, an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0107] The diagnostic kits of the present invention may further comprise one or more other component compositions, solutions or devices suitable for the analytical method.

[0108] For example, the diagnostic kit of the present invention may further include essential elements required for performing a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes a primer pair specific to a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. In addition, a primer specific to the nucleic acid sequence of a control gene may be included. In addition, the reverse transcription polymerase reaction kit may include a test tube or other suitable container, a reaction buffer (varies in pH and magnesium concentration), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor, DEPC water, sterile water, etc.

[0109] The diagnostic kit of the present invention can also include essential elements required for performing a DNA chip. The DNA chip kit can include a substrate to which cDNA or oligonucleotides corresponding to a gene or a fragment thereof are attached, and reagents, preparations, enzymes, and the like for producing fluorescently labeled probes. Furthermore, the substrate can include cDNA or oligonucleotides corresponding to a control gene or a fragment thereof.

[0110] The diagnostic kit of the present invention may include essential elements required for performing ELISA. The ELISA kit includes an antibody specific to the protein. The antibody has high specificity and affinity for the marker protein and has almost no cross-reactivity with other proteins, and is a monoclonal antibody, a polyclonal antibody, or a recombinant antibody. Furthermore, the ELISA kit may include an antibody specific to a control protein. In addition, the ELISA kit may include reagents capable of detecting the bound antibody, such as a labeled secondary antibody, chromophores, an enzyme (e.g., conjugated with the antibody) and its substrate or other substances capable of binding to the antibody.

[0111] According to yet another embodiment of the present invention, there is provided a method for measuring the expression level of Lrig-1 protein present in a biological sample of an individual of interest using the binding molecule provided by the present invention, thereby providing information regarding the diagnosis of a nervous system disease.

[0112] In the present invention, the "target individual" refers to an individual for whom it is uncertain whether or not he or she will develop a cranial nervous system disease, particularly a neurodegenerative disease or a neuroinflammatory disease, and who has a high probability of developing the disease.

[0113] In the present invention, the "biological sample" may be any material obtained from or derived from an individual, tissue, cell, whole blood, serum, plasma, tissue autopsy samples (brain, skin, lymph nodes, spinal cord, etc.), cell culture supernatant, ruptured eukaryotic cells, and bacterial expression systems, but may preferably be regulatory T cells.

[0114] In the present invention, the method for measuring or comparatively analyzing the expression level of the Lrig-1 protein includes, but is not limited to, protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radial immunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, complement fixation analysis, two-dimensional electrophoresis analysis, liquid chromatography-Mass Spectrometry (LC-MS), LC-MS / MS (liquid chromatography-Mass Spectrometry / Mass Spectrometry), Western blotting, or ELISA (enzyme linked immunosorbent assay).

[0115] In the present invention, the method may include a step of predicting that there is a high possibility of developing a neurodegenerative disease or a neuroinflammatory disease when the expression level of the Lrig-1 protein measured in a biological sample of an individual of interest is increased or decreased compared to a normal control group.

[0116] In the present invention, the cranial nervous system disease to be diagnosed by the diagnostic composition, the diagnostic kit, or the information providing method may be a neurodegenerative disease or a neuroinflammatory disease, and specific examples thereof include stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Niemann-Pick disease, multiple sclerosis, prion disease, Creutzfeldt-Jakob disease, frontotemporal dementia, dementia with Lewy bodies, amyotrophic lateral sclerosis, paraneoplastic syndrome, corticobasal degeneration, multiple system atrophy, progressive supranuclear palsy, autoimmune diseases of the nervous system, spinocerebellar ataxia, and the like. The therapeutic agent may be selected from the group consisting of, but is not limited to, ataxia, inflammatory and neuropathic pain, cerebrovascular disease, spinal cord injury and tauopathy. Effect of the Invention

[0117] The binding molecules provided by the present invention can not only specifically prevent, ameliorate or treat brain and nervous system diseases such as various neurodegenerative or neuroinflammatory diseases, but also provide a method for diagnosing the same. [Brief description of the drawings]

[0118] [Figure 1] FIG. 1 shows the structure of Lrig-1 protein according to one embodiment of the present invention. [Diagram 2] FIG. 1 shows the structure of Lrig-1 protein according to one embodiment of the present invention. [Diagram 3] FIG. 2 shows the results of predicting the epitope of Lrig-1 protein according to one embodiment of the present invention. [Figure 4] FIG. 2 shows the results of predicting the epitope of Lrig-1 protein according to one embodiment of the present invention. [Diagram 5] A graph showing the expression level of Lrig-1 mRNA according to one embodiment of the present invention. [Figure 6] A graph showing the expression level of Lrig-1 mRNA according to one embodiment of the present invention. [Figure 7] A graph showing the expression level of Lrig-1 mRNA according to one embodiment of the present invention. [Figure 8] A figure showing the expression levels of Lrig-1, Lrig-2 and Lrig-3 mRNA according to one embodiment of the present invention. [Figure 9] FIG. 1 shows a comparison of the expression levels of Lrig-1 protein in regulatory T cells and non-regulatory T cells according to one embodiment of the present invention. [Figure 10] FIG. 1 shows expression of Lrig-1 protein on the surface of regulatory T cells according to one embodiment of the present invention. [Figure 11] FIG. 1 shows the results of analyzing the binding affinity of an Lrig-1 protein-specific monoclonal antibody according to one embodiment of the present invention to Lrig-1 protein. [Figure 12] 1 shows an experimental design diagram for confirming the therapeutic effect of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention on multiple sclerosis. [Figure 13] FIG. 13 shows the results of an analysis of the therapeutic effect of multiple sclerosis after administration of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention. [Figure 14] FIG. 13 shows the results of an analysis of the therapeutic effect of Alzheimer's disease after administration of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention. [Figure 15] FIG. 13 shows the results of an analysis of the therapeutic effect of Alzheimer's disease after administration of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention. [Figure 16] FIG. 13 shows the results of an analysis of the therapeutic effect of Alzheimer's disease after administration of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention. [Figure 17] FIG. 13 shows the results of an analysis of the therapeutic effect of Alzheimer's disease after administration of a monoclonal antibody specific to Lrig-1 protein according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0119] The present invention relates to a composition for preventing, ameliorating, or treating a brain and nervous system disease, which comprises as an active ingredient a binding molecule that specifically binds to the Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present in regulatory T cells (Treg cells).

[0120] wherein the binding molecule is

[0121] A heavy chain variable region comprising: a heavy chain CDR1 consisting of the amino acid sequence represented by SEQ ID NO:5 or 13; a heavy chain CDR2 consisting of the amino acid sequence represented by SEQ ID NO:6 or 14; and a heavy chain CDR3 consisting of the amino acid sequence represented by SEQ ID NO:7 or 15; and

[0122] The binding molecule may comprise a light chain variable region including: a light chain CDR1 represented by the amino acid sequence represented by SEQ ID NO: 8 or 16; a light chain CDR2 represented by the amino acid sequence represented by SEQ ID NO: 9 or 17; and a light chain CDR3 represented by the amino acid sequence represented by SEQ ID NO: 10 or 18. EXAMPLES

[0123] The present invention will be described in more detail with reference to the following examples. It will be obvious to those skilled in the art that these examples are merely intended to more specifically illustrate the present invention, and that the scope of the present invention is not limited to these examples according to the gist of the present invention.

[0124] Working Example [Preparation Example 1] Cultivation of T cell subtypes To confirm whether Lrig-1 protein is expressed only in regulatory T cells (Treg), T cell subtypes Th0, Th1, Th2, Th17, and iTreg were prepared. The iTreg is different from naturally isolated nTreg, and refers to cells whose differentiation was artificially induced in a medium containing the following composition.

[0125] To determine the subtypes of T cells, naive T cells were first isolated from the mouse spleens and then cultured in RPMI1640 (Invitrogen Gibco, Grand Island, NY) nutrient medium containing 10% fetal bovine serum (FBS; hyclone, logan, UT) and the components listed in Table 1 below for 72 hours in a 37°C, 5% CO2 incubator to induce differentiation into each type of cell.

[0126] [Table 1]

[0127] [Example 1] Analysis of Lrig-1 structure In order to generate antibodies specific to Lrig-1 protein, a surface protein of regulatory T cells, the three-dimensional structure of the extracellular domain of Lrig-1 protein was predicted.

[0128] First, to confirm the structure of the extracellular domain (ECD) of the Lrig-1 protein for predicting the epitope base sequence, the three-dimensional structure was predicted using Uniprot (http: / / www.uniprot.org) and RCSB Protein Data Bank (http: / / www.rcsb.org / pdb) tools, and the results are shown in Figures 1 and 2.

[0129] As shown in Figure 1, the Lrig-LRR domain (amino acid sequence 41 to 494) in the extracellular domain of the Lrig-1 protein contained a total of 15 leucine-rich regions, LRR1 to LRR15. Each of the LRR domains was composed of 23 to 27 amino acids and contained 3 to 5 leucines.

[0130] Furthermore, as shown in FIG. 2, the extracellular domain of the Lrig-1 protein contained three immunoglobulin-like domains in the amino acid sequence 494 to 781 of the Lrig-1 protein.

[0131] [Example 2] Prediction of Lrig-1 epitope amino acid sequence The base sequence was predicted using the Ellipro server (http: / / tools.iedb.org / ellipro / ), which is an epitope prediction software based on the structure of the Lrig-1 protein. The Ellipro search engine was used because it is known to be the most reliable of the currently available algorithms for predicting antigenic determinants.

[0132] After inputting the extracellular domain analyzed in Example 1 into the antigenic determinant prediction software, the predicted contiguous or discontinuous amino acid sequences of the predicted antigenic determinants are shown in FIGS.

[0133] As shown in Figures 3 and 4, a total of 22 amino acid sequences of continuous antigenic determinants were predicted, and a total of 8 amino acid sequences of discontinuous antigenic determinants were predicted.

[0134] [Production Examples 1 to 8] Preparation of monoclonal antibodies specific to Lrig-1 protein The present invention has produced an antibody specific to the Lrig-1 protein. This antibody was produced not by determining a specific antigenic determinant, but by producing an antibody capable of binding to any site on the Lrig-1 protein.

[0135] To prepare the antibody, cells expressing Lrig-1 protein were prepared. More specifically, the DNA fragment corresponding to SEQ ID NO: 2 and pcDNA (hygro) were cleaved with a cleavage enzyme, then cultured at 37°C for ligation to prepare pcDNA into which the DNA sequence of Lrig-1 protein was inserted. The prepared pcDNA into which SEQ ID NO: 2 was inserted was introduced into L cells by transfection to allow the expression of Lrig-1 protein on the surface of the L cells.

[0136] A total of eight heavy and light chains were selected from the Human scFv library for light and heavy chain amino acid sequences capable of binding to Lrig-1 expressed on the cell surface.

[0137] The selected heavy and light chain amino acid sequences were fused to the MLG2a Fc region to produce monoclonal antibodies, the sequences of which are shown in Table 2 below.

[0138] [Table 2-1]

[0139] [Table 2-2]

[0140] [Table 2-3]

[0141] [Table 2-4]

[0142] [Table 2-5]

[0143] [Example 3] Confirmation of specific expression of Lrig-1 mRNA in regulatory T cells We examined whether Lrig-1 protein could act as a specific biomarker for regulatory T cells.

[0144] For this validation, CD4 cells were extracted from mouse spleens using magnet-activated cell sorting (MACS) with CD4 beads. + T cells were isolated. Thereafter, regulatory T cells (CD4 + CD25 + T) cells and non-regulatory T (CD4 + CD25 - T) cells were isolated. After extracting mRNA from each of the cells and the cells differentiated in Preparation Example 1 using Trizol, gDNA was removed from the genomic RNA using a gDNA extraction kit (Qiagen) according to the protocol provided by the company. The gDNA-removed mRNA was synthesized into cDNA using the BDsprint cDNA synthesis kit (Clonetech).

[0145] To quantitatively confirm the expression level of Lrig-1 mRNA using the cDNA, real-time polymerase chain reaction (real-time PCR) was performed.

[0146] The real-time polymerase chain reaction was performed using SYBR Green (Molecular Probes) according to the protocol provided by the company under the conditions of 95°C for 3 minutes, 61°C for 15 seconds, and 72°C for 30 seconds, for 40 cycles, using the primers in Table 3 below. The relative gene expression levels were calculated using the ΔCT method and normalized using HPRT. The results are shown in Figures 5 to 8.

[0147] [Table 3]

[0148] As shown in Figure 5, non-regulatory T (CD4 + CD25 - T) cells, compared with regulatory T (CD4 + CD25 + It can be seen that Lrig-1 expression was 18.1 times higher in naturally isolated T cells. This was about 10 times higher than the previously known regulatory T cell markers Lag3 and Ikzf4. Furthermore, as shown in Figures 6 and 7, Lrig-1 mRNA expression was significantly higher in naturally isolated T cells (nTreg) than in other types of immune cells, particularly in induced T cells (iTreg).

[0149] Furthermore, as shown in FIG. 8, among Lrig family members Lrig-1, Lrig-2 and Lrig-3, expression of Lrig-1 was the highest.

[0150] From the above results, it can be seen that the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells, particularly in naturally occurring regulatory T cells.

[0151] [Example 4] Confirmation of specific expression of Lrig-1 protein in regulatory T cells We confirmed whether the Lrig-1 protein expressed from Lrig-1 mRNA was specifically expressed only in regulatory T cells.

[0152] Using FOXP3-RFP knock-in mice, in which RFP (red fluorescence protein) is bound to the FOXP3 promoter, a transcription factor specific to regulatory T cells, CD4 cells were sorted from the spleen of the mice using a magnet-activated cell sorting (MACS) machine with CD4 beads. + T cells were isolated. Thereafter, regulatory T cells (CD4 + RFP + T) cells and non-regulatory T (CD4 + RFP - The cells were stained with the purchased Lrig-1 antibody and the negative control group was stained with an isotype antibody, and the expression level of Lrig-1 was measured using a fluorescence activated cell sorter. The results are shown in FIG.

[0153] As shown in Figure 9, non-regulatory T cells, represented by the dotted line, showed Lrig-1 expression levels almost the same as those of the negative control group, whereas regulatory T cells contained a large number of cells with high Lrig-1 expression levels.

[0154] From the above results, it can be seen that the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells.

[0155] [Example 5] Confirmation of specific expression of Lrig-1 protein on the surface of regulatory T cells In order for the Lrig-1 protein to be a target for cell therapy, it must be expressed on the surface of regulatory T cells for effective targeted therapy, so we confirmed whether the Lrig-1 protein is expressed on the surface.

[0156] Each differentiated T cell subtype from Preparatory Example 1 was stained with anti-CD4-APC and anti-Lrig-1-PE antibodies, and the expression level of Lrig-1 on the cell surface was measured using a Fluorescence-Activated Cell Sorter (FACS). The results are shown in Figure 10.

[0157] As shown in Figure 10, Lrig-1 was expressed at levels of 0.77 to 15.3 in activated T cells, Th1 cells, Th2 cells, Th17 cells, and naive T cells, whereas it was highly expressed at 83.9 in differentiated T cells (iTreg).

[0158] From the above results, it can be seen that the Lrig-1 protein according to the present invention is not only specifically expressed in regulatory T cells (Treg) cells, but is particularly highly expressed on the surface of Treg cells.

[0159] [Example 6] Evaluation of the binding ability of the antibody according to the present invention to Lrig-1 protein In order to confirm whether the monoclonal antibodies of the present invention prepared in Preparation Examples 1 to 8 can effectively recognize Lrig-1, each of the antibodies prepared in Preparation Examples 1 to 8 was bound to L cells stably expressing Lrig-1, and then a secondary antibody capable of recognizing mouse antibodies and conjugated with eFlour670 was added, and the binding strength of the monoclonal antibodies to the Lrig-1 protein was analyzed using FACS, and the results are shown in Figure 11.

[0160] As shown in Figure 11, it was confirmed that the Lrig-1 protein-specific monoclonal antibodies according to the present invention (GTC110-04 and GTC210-01) effectively recognized and bound to the Lrig-1 protein present on the surface of L cells.

[0161] [Example 7] Evaluation of therapeutic ability of the antibody according to the present invention for multiple sclerosis 1. Creation of Multiple Sclerosis Mouse Model To prepare a mouse model of multiple sclerosis, an experiment was performed according to the experimental design diagram shown in FIG. 12. On day 0, 7-week-old C57BL / 6 mice were subcutaneously injected with MOG peptide and intraperitoneally injected with Mycobacterium tuberculosis toxin. On day 2, the mice were injected with Mycobacterium tuberculosis toxin again to boost their immune system. On day 9, the mice were administered with the GTC210-01 antibody prepared in Preparation 1, and anti-IL-17a antibody was administered as a positive control.

[0162] 2. Evaluation of therapeutic effects on multiple sclerosis From the 7th day, the therapeutic effect on multiple sclerosis in mice was evaluated by calculating a clinical score according to the following evaluation criteria, and the results are shown in FIG.

[0163] <Evaluation criteria> 0 points: No symptoms, functioning normally 1 point: Part of the end of the tail is paralyzed and hangs down 2 points: The entire tail is paralyzed and drooping 3 points: One of the hind legs is partially paralyzed, but still able to respond to stimuli 4 points: One of the hind legs is completely paralyzed and unresponsive, dragging the leg 5 points: All hind legs are paralyzed and the animal drags itself along using its front legs 6 points: Difficulty dragging with front legs, but front legs still responsive to stimulation 7 points: Mouse cannot move, but one front paw responds to stimulation 8 points: Mouse is immobile and neither front paw responds to stimulation 9 points: Mouse is immobile and breathing is irregular 10 points: Mouse is dead

[0164] As a result, when the antibody according to the present invention was administered to an induced multiple sclerosis model, it was confirmed that multiple sclerosis was significantly treated, and in particular, it was confirmed that recovery was achieved to a level similar to that of normal mice by the 22nd and 23rd days.

[0165] [Example 8] Evaluation of the therapeutic ability of the antibody according to the present invention for Alzheimer's To evaluate the therapeutic ability of the antibody according to the present invention against Alzheimer's disease, the groups shown in Table 4 below were designed. Specifically, 7-month-old Alzheimer's-induced 5xFAD mice were intravenously injected with GTC110-04 antibody and GTC210-01 antibody at 10 mpk each for one month. 8-month-old Alzheimer's mice were subcutaneously injected with glatiramer acetate (GA, Copaxone®) at 100 μg for three weeks as a positive control group.

[0166] [Table 4]

[0167] 1. Y-maze test The Y-maze test used a maze frame with three identical arms, 40 cm long (wall height 15 cm), arranged at a 120 degree angle. This is a behavioral experiment that utilizes the instinctive exploration behavior of rodents, focusing on the high possibility of exploring new areas. The higher the memory score, the more the rat remembered the arm it had just explored and did not enter the same arm. Each rat was given 8 minutes of exploration time, and the final results are shown in Figure 14 as spontaneous alteration (%). The spontaneous alteration (%) score was calculated using the following formula 1. Here, the behavioral pattern analysis was performed using SMART VIDEO TRACKING Software (Panlab, USA).

[0168] [Formula 1] Spontaneous alteration(%)= The number of triplet / (total arm entry-2)

[0169] As a result, in this experiment, which measured the relative frequency with which experimental animals grasped peripheral cues and entered the maze sequentially, it was confirmed that when Alzheimer's-induced mice were administered the antibodies according to the present invention (GTC210-01 and GTC110-04 antibodies), they had spontaneous alternation values ​​at a similar level to the normal control group.

[0170] 2. Novel Object Recognition The novel object recognition test was performed to evaluate memory ability using two different objects in a 40 cm × 40 cm acrylic cage. After getting used to the inside of the acrylic cage, the two objects were placed in fixed positions and allowed to be recognized freely, and the time spent exploring each object was measured. A 24-hour delay was given to each individual, and only one object was replaced with another object in the same position. At this time, the longer the exploration time after recognizing the replaced object as a novel object, the better the memory is judged to be. If the object from 24 hours ago is not remembered, it is possible that the new object cannot be distinguished from the existing object and that both objects are explored equally. The rats were allowed to explore freely for a total of 10 minutes, and the results are shown in Figure 15 as a preference index (novel object exploring time / total exploring time). Analysis was performed using SMART VIDEO TRACKING Software (Panlab, USA).

[0171] As a result, the preference index for the novel object was measured as 0.51±0.06 in the normal control group (G1) and 0.42±0.04 in the G2 group (vehicle), and the preference index was observed to be lower in the Alzheimer's-induced group (G2) than in the normal control group (G1). However, it was confirmed that when the antibodies according to the present invention (GTC210-01 and GTC110-04 antibodies) were administered, the preference index increased to a level higher than that of the normal control group.

[0172] 3. Water maze test The underwater maze test was performed based on the method devised by Morris. A stainless steel pool (diameter 90 cm, height 50 cm) was filled with water (22 ± 1°C) and the water surface was set to a height of 30 cm. A hidden platform (diameter 5 cm) was placed 1 cm below the water surface. Three to four practice sessions were conducted on the first day of evaluation. The total swimming time for each animal was set to 60 seconds, and animals that found the platform within 60 seconds were made to stay on the platform for 10 seconds to induce memory. Animals that could not find the platform after 60 seconds were artificially guided to the platform and made to stay on the platform for 10 seconds, with an escape time of 60 seconds. After the swimming practice, a probe test was conducted on the sixth day to measure spatial perception ability by calculating the percentage of time spent in the four quadrants where the platform was in relation to the total swimming time (Percentage of time in SW area, %) and the time it took to reach the platform location (Latency to target, sec). The results are shown in Figures 16 and 17. Analysis was performed using SMART VIDEO TRACKING Software (Panlab, USA). The criterion for exclusion from the interpretation of the results was set to individuals that did not explore by swimming but only turned around a specific area.

[0173] As a result, when the antibodies according to the present invention (GTC210-01 and GTC110-04 antibodies) were administered, the time spent in the four quadrants where the platform was located, calculated as a percentage of the total swimming time (Percentage of time in SW area, %), significantly increased, and it was confirmed that the time it took to reach the position of the platform (Latency to target, sec) significantly decreased to a level similar to that of the normal control group.

[0174] Through these experiments, it was confirmed that the antibody according to the present invention has the effect of preventing, improving or treating diseases of the central nervous system.

[0175] Although the present invention has been described in detail above, the scope of the invention is not limited thereto, and it will be obvious to those having ordinary skill in the art that various modifications and variations are possible within the scope that does not deviate from the technical idea of ​​the invention described in the claims.

[0176] [Industrial Applicability] The present invention relates to a composition for preventing or treating a brain and nervous system disease.

[0177]

Claims

1. A pharmaceutical composition for preventing or treating a brain and nervous system disease, comprising as an active ingredient a binding molecule that specifically binds to Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present on the surface of regulatory T cells (Treg cells), the binding molecule is an antibody or an antigen-binding fragment thereof; The therapeutic pharmaceutical composition, wherein the brain and nervous system disease is Alzheimer's disease or multiple sclerosis.

2. The pharmaceutical composition according to claim 1, wherein the Lrig-1 protein consists of the amino acid sequence represented by SEQ ID NO: 1 or 3.

3. The pharmaceutical composition of claim 1, wherein the Lrig-1 protein is encoded by a polynucleotide represented by SEQ ID NO: 2 or 4.

4. The antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising a heavy chain CDR1 consisting of the amino acid sequence represented by SEQ ID NO:5 or 13, a heavy chain CDR2 consisting of the amino acid sequence represented by SEQ ID NO:6 or 14, and a heavy chain CDR3 consisting of the amino acid sequence represented by SEQ ID NO:7 or 15; and A light chain variable region comprising a light chain CDR1 represented by the amino acid sequence represented by SEQ ID NO:8 or 16, a light chain CDR2 represented by the amino acid sequence represented by SEQ ID NO:9 or 17, and a light chain CDR3 represented by the amino acid sequence represented by SEQ ID NO:10 or 18. The pharmaceutical composition of claim 1 , comprising:

5. The antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO:5, a heavy chain CDR2 represented by SEQ ID NO:6, and a heavy chain CDR3 represented by SEQ ID NO:7; and (b) a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO: 13, a heavy chain CDR2 represented by SEQ ID NO: 14, and a heavy chain CDR3 represented by SEQ ID NO: 15; a heavy chain variable region selected from the group consisting of: (c) a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO: 8, a light chain CDR2 represented by SEQ ID NO: 9, and a light chain CDR3 represented by SEQ ID NO: 10; and (d) a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO: 16, a light chain CDR2 represented by SEQ ID NO: 17, and a light chain CDR3 represented by SEQ ID NO:

18. A light chain variable region selected from the group consisting of The pharmaceutical composition of claim 1 , comprising:

6. The antibody or antigen-binding fragment thereof comprises: (1) A binding molecule comprising a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO:5, a heavy chain CDR2 represented by SEQ ID NO:6, and a heavy chain CDR3 represented by SEQ ID NO:7; and a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO:8, a light chain CDR2 represented by SEQ ID NO:9, and a light chain CDR3 represented by SEQ ID NO:10; and (2) A binding molecule comprising a heavy chain variable region comprising a heavy chain CDR1 represented by SEQ ID NO: 13, a heavy chain CDR2 represented by SEQ ID NO: 14, and a heavy chain CDR3 represented by SEQ ID NO: 15; and a light chain variable region comprising a light chain CDR1 represented by SEQ ID NO: 16, a light chain CDR2 represented by SEQ ID NO: 17, and a light chain CDR3 represented by SEQ ID NO:

18. The pharmaceutical composition of claim 1, selected from the group consisting of:

7. The antibody or antigen-binding fragment thereof comprises: A heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or 19; and A light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or 20 The pharmaceutical composition of claim 1 , comprising:

8. The antibody or antigen-binding fragment thereof comprises: A binding molecule comprising a heavy chain variable region represented by SEQ ID NO:11 and a light chain variable region represented by SEQ ID NO:12; and The pharmaceutical composition of claim 1 , selected from the group consisting of binding molecules comprising a heavy chain variable region represented by SEQ ID NO: 19 and a light chain variable region represented by SEQ ID NO:

20.

9. The pharmaceutical composition of claim 1 , wherein the antibody or antigen-binding fragment thereof further comprises an Fc region or a constant region.

10. 10. The pharmaceutical composition of claim 9, wherein the Fc region is an Fc region of an IgA, IgD, IgE, IgM, IgG1, IgG2, IgG3 or IgG4 antibody, or a hybrid Fc region.

11. The pharmaceutical composition of claim 1 , wherein the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 23, 24, 26, 27, 28 and 29.

12. The pharmaceutical composition of claim 1 , wherein the antibody or antigen-binding fragment thereof further comprises a light chain constant region consisting of the amino acid sequence represented by SEQ ID NO: 22 or 25.

13. The antibody or antigen-binding fragment thereof comprises: A heavy chain constant region consisting of the amino acid sequence represented by SEQ ID NO:21; and The pharmaceutical composition of claim 1 , further comprising a light chain constant region consisting of the amino acid sequence represented by SEQ ID NO:

22.

14. The antibody or antigen-binding fragment thereof comprises: A heavy chain constant region consisting of the amino acid sequence represented by SEQ ID NO: 23, 24, 26, 27 or 28; and The pharmaceutical composition of claim 1, further comprising a light chain constant region consisting of the amino acid sequence represented by SEQ ID NO:

25.

15. The pharmaceutical composition of claim 1 , wherein the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region consisting of the amino acid sequence represented by SEQ ID NO:

29.

16. The antibody or antigen-binding fragment thereof comprises: A binding molecule comprising a heavy chain represented by SEQ ID NO: 30 and a light chain represented by SEQ ID NO: 31; A binding molecule comprising a heavy chain represented by SEQ ID NO:32 and a light chain represented by SEQ ID NO:33; and A binding molecule comprising a heavy chain represented by SEQ ID NO:34 and a light chain represented by SEQ ID NO:

35. The pharmaceutical composition of claim 1, which is selected from the group consisting of:

17. 2. The pharmaceutical composition of claim 1, wherein the antibody or antigen-binding fragment thereof is a chimeric antibody, a humanized antibody, a bivalent antibody, a bispecific molecule, a minibody, a domain antibody, a bispecific antibody, a unibody, a diabody, a triabody, a tetrabody, or a fragment thereof.

18. A nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof that specifically binds to Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present on the surface of regulatory T cells (Treg cells); an expression vector into which the nucleic acid molecule has been inserted; or A host cell line transfected with the expression vector A pharmaceutical composition for preventing or treating a brain and nervous system disease, comprising as an active ingredient: The cranial nervous system disease is Alzheimer's disease or multiple sclerosis, The antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a heavy chain CDR1 having the amino acid sequence represented by SEQ ID NO:5 or 13; a heavy chain CDR2 having the amino acid sequence represented by SEQ ID NO:6 or 14; and a heavy chain CDR3 having the amino acid sequence represented by SEQ ID NO:7 or 15; and a light chain variable region comprising a light chain CDR1 having the amino acid sequence represented by SEQ ID NO:8 or 16; a light chain CDR2 having the amino acid sequence represented by SEQ ID NO:9 or 17; and a light chain CDR3 having the amino acid sequence represented by SEQ ID NO:10 or 18. Therapeutic pharmaceutical compositions.

19. A pharmaceutical composition for preventing or treating a brain and nervous system disease, comprising an antibody-drug conjugate (ADC) as an active ingredient, The cranial nervous system disease is Alzheimer's disease or multiple sclerosis, The antibody in the antibody-drug conjugate is It specifically binds to Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present on the surface of regulatory T cells (Treg cells), A heavy chain variable region comprising: a heavy chain CDR1 consisting of the amino acid sequence represented by SEQ ID NO:5 or 13; a heavy chain CDR2 consisting of the amino acid sequence represented by SEQ ID NO:6 or 14; and a heavy chain CDR3 consisting of the amino acid sequence represented by SEQ ID NO:7 or 15; and A light chain variable region including a light chain CDR1 represented by the amino acid sequence represented by SEQ ID NO: 8 or 16; a light chain CDR2 represented by the amino acid sequence represented by SEQ ID NO: 9 or 17; and a light chain CDR3 represented by the amino acid sequence represented by SEQ ID NO: 10 or 18. Therapeutic pharmaceutical compositions.

20. A composition for diagnosing a brain and nervous system disease, comprising an antibody or an antigen-binding fragment thereof, The cranial nervous system disease is Alzheimer's disease or multiple sclerosis, The antibody or antigen-binding fragment thereof comprises: It specifically binds to Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present on the surface of regulatory T cells (Treg cells), A heavy chain variable region comprising: a heavy chain CDR1 consisting of the amino acid sequence represented by SEQ ID NO:5 or 13; a heavy chain CDR2 consisting of the amino acid sequence represented by SEQ ID NO:6 or 14; and a heavy chain CDR3 consisting of the amino acid sequence represented by SEQ ID NO:7 or 15; and A light chain variable region including a light chain CDR1 represented by the amino acid sequence represented by SEQ ID NO: 8 or 16; a light chain CDR2 represented by the amino acid sequence represented by SEQ ID NO: 9 or 17; and a light chain CDR3 represented by the amino acid sequence represented by SEQ ID NO: 10 or 18. Diagnostic compositions.

21. A diagnostic kit for a cranial nervous system disease, comprising the diagnostic composition according to claim 20.

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