Anti-cd300c antibody or antigen binding fragment of the same and uses thereof for preventing or treating degenerative brain diseases

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of degenerative brain disease comprising one or more of an anti-CD300c antibody and an antigen-binding fragment thereof; a method for the prevention or treatment of degenerative brain disease using the same; a feed composition for the prevention or improvement of degenerative brain disease comprising one or more of an anti-CD300c antibody and an antigen-binding fragment thereof; a food composition; and the use of the anti-CD300c antibody and the antigen-binding fragment thereof for the prevention or treatment of degenerative brain disease.

Description

Technology Field

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of degenerative brain disease comprising one or more of an anti-CD300c antibody and an antigen-binding fragment thereof; a method for the prevention or treatment of degenerative brain disease using the same; a feed composition or food composition for the prevention or improvement of degenerative brain disease comprising one or more of an anti-CD300c antibody and an antigen-binding fragment thereof; and the use of the anti-CD300c antibody and / or the antigen-binding fragment thereof for the prevention or treatment of degenerative brain disease. Background Technology

[0003] Alzheimer's disease (AD) is a representative example of a degenerative brain disease characterized by the progressive loss of cognitive and memory functions and neurodegeneration of the central nervous system, accounting for approximately 50% of all forms of dementia. AD is characterized by the presence of amyloid plaques, neurofibrillary tangles, synaptic loss, and selective neurogenic death. Amyloid plaques are caused by abnormal levels of extracellular amyloid beta-peptides, while neurofibrillary tangles are associated with the presence of intracellular hyperphosphorylated tau protein. Symptoms are characterized by forgetfulness, along with dysphasia, dyspraxia, and agnosia (the inability to recognize objects, people, sounds, shapes, or scents), which contribute to their association with cortical association sites. The incidence of AD increases particularly after age 65 and is rapidly rising worldwide as the population ages.

[0004] While commercially available AD treatments help improve cognitive function, they are primarily composed of AChE inhibitors and NMDA receptor antagonists aimed at temporary symptom relief. Aducanumab, a monoclonal antibody drug targeting amyloid beta, received FDA approval in 2021; unlike previous symptom-relief agents, it is a treatment that targets the underlying pathophysiology, although there is controversy regarding its efficacy and side effects. Additionally, research has been reported on treating Alzheimer's disease by targeting the PD-1 / PD-L1 signaling pathway (Non-patent Literature 1). This study suggested the potential of immunological methods, indicating that blocking PD-1 / PD-L1 signaling increased monocyte-derived macrophages in the brain parenchyma, consequently leading to a reduction in plaques and brain lesions. Although developing AD treatments is difficult and has a very low success rate, the acceleration of the aging society, the increase in AD patients, and the growing socioeconomic burden caused by AD necessitate the development of various drugs for fundamental treatment rather than just symptom relief.

[0005] Meanwhile, CD300c (CD300 antigen-like family member C) protein is a protein encoded by the CD300c gene in humans, and it is present on the surface of various tumor cells and monocytes. It has been revealed that inhibiting the activity or expression of CD300c protein can reduce the proliferation of cancer cells by improving immune function (e.g., T cell activation) through this (Patent Document 1). Prior art literature

[0007] Korean Patent Publication No. 10-2021-0060355

[0008] 1. Rosenzweig, N., Dvir-Szternfeld, R., Tsitsou-Kampeli, A., Keren-Shaul, H., Ben-Yehuda, H., Weill-Raynal, P., Cahalon, L., Kertser, A., Baruch, K., Amit, I., Weiner, A., & Schwartz, M. PD-1 / PD-L1 checkpoint blockade harnesses monocyte-derived macrophages to combat cognitive impairment in a tauopathy mouse model. Nature Communications, 10(1) (2019) The problem to be solved

[0009] The present invention aims to solve all of the aforementioned problems.

[0010] Another objective of the present invention is to provide an antibody drug for the prevention or treatment of degenerative brain diseases.

[0011] Another objective of the present invention is to provide a composition for the prevention or treatment of degenerative brain diseases.

[0012] Another objective of the present invention is to provide a method for preventing or treating degenerative brain diseases.

[0013] Another objective of the present invention is to provide the use of the anti-CD300c antibody and its antigen-binding fragment for the prevention or treatment of degenerative brain diseases. means of solving the problem

[0015] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may be applied to other descriptions and embodiments. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions provided below. Additionally, numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated by reference into this specification in their entirety to more clearly explain the level of the art to which the present invention pertains and the content of the present invention.

[0017] One aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of a degenerative brain disease, comprising one or more of an anti-CD300c antibody and an antigen-binding fragment thereof as an active ingredient.

[0018] In one embodiment, the antibody of the present invention may have the ability to promote differentiation from monocytes into MDM (Monocyte-derived macrophage) or M1 macrophage, and may have an increased ability compared to general therapeutic agents for neurodegenerative diseases and immunotherapies, but is not limited thereto. In the present invention, MDM may be M1-MDM.

[0019] In the present invention, the anti-CD300c antibody and its antigen-binding fragment refer to antigen-binding molecules capable of specifically binding to the CD300c protein, and may, but are not limited to, inhibit the reaction of CD300c.

[0020] In the present invention, the term "CD300c" is used interchangeably with "CD300c protein" or "CD300c antigen," and is a type of CD300 family cell surface cell involved in immune regulation that may be expressed in antigen-presenting cells, cancer cells, or immune cells, but is not limited thereto.

[0021] In the present invention, the term "antibody" refers to a proteinaceous molecule capable of specifically recognizing an antigenic site, comprising an immunoglobulin or a part thereof that is immunologically reactive with a specific antigen. The antibodies of the present invention include polyclonal antibodies, monoclonal antibodies, whole antibodies, and antibody fragments. Furthermore, the antibodies of the present invention include antibodies from mice, humans, rabbits, and rats, without being limited to their origin. The antibodies of the present invention include animal antibodies, chimeric antibodies (e.g., humanized murine antibodies), humanized antibodies, multispecific antibodies, bivalent or bispecific molecules (e.g., bispecific antibodies), diabodies, triabodidies, tetrabodies, minibodies, and antibody fusions (e.g., fusions of antibodies and (poly)peptides or fusions of antibodies and compounds). The antibodies of the present invention further comprise short-chain antibodies having a binding function to FcRn (neonatal Fc receptor), scaffolds, derivatives of antibody constant regions, and artificial antibodies based on protein scaffolds. The total antibody has a structure having two full-length light chains (LC) and two full-length heavy chains (HC), and each light chain may be connected to the heavy chain by a disulfide bond. The total antibody includes any antibody such as IgA, IgD, IgE, IgM, and IgG, wherein IgG includes subtypes IgG1, IgG2, IgG3, and IgG4. Such antibodies may be prepared by cloning each gene into an expression vector according to a conventional method to obtain a protein encoded by said marker gene, and from the obtained protein by a conventional method, but are not limited thereto.

[0022] In the present invention, the prefix "anti-" indicates that, when associated with an antigen, the corresponding antibody is reactive with that antigen. Antibodies reactive with a specific antigen may be produced by synthetic and / or recombinant methods, such as the selection of a recombinant antibody library from a phage or similar vector, or by the immunization of animals using an antigen or antigen-coding nucleic acid, but are not limited thereto.

[0023] In one embodiment, the anti-CD300c antibody of the present invention may mean an antibody that is reactive with CD300c.

[0025] In the present invention, the term “fragment” or “antibody fragment” refers to any part of an antibody, and may include scFv, dsFv, Fab, Fab’, F(ab’)2, Fc, Fd, sdAb, nanobody, and combinations thereof, and may include an antibody fragment, and may include an antigen recognition site, but is not limited thereto.

[0026] In the present invention, the term "antigen-binding fragment" refers to a fragment possessing an antigen-binding function. In the present invention, the antigen-binding fragment may be a fragment comprising a site capable of recognizing an antigenic site.

[0027] In the present invention, Fd refers to the heavy chain portion included in the Fab fragment. In the present invention, Fab has a structure having a variable region 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. 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 the smallest antibody fragment having only the heavy chain variable region and the light chain variable region. In a double disulfide Fv (dsFv), the heavy chain variable region and the light chain variable region are connected by disulfide bonds, and in a single chain variable fragment (scFv), the heavy chain variable region and the light chain variable region are generally connected by covalent bonds through a peptide linker. A double disulfide single chain Fv may be a single chain Fv in which the heavy chain variable region and the light chain variable region are additionally connected by disulfide bonds. The above sdAb and nanobody are single variable domain antibody fragments, and may include, for example, antibody fragments produced by protein hydrolysis or genetic recombination technology among heavy chain antibodies containing a naturally occurring single variable domain (VH) and two constant domains (CH2 and CH3), and single domain antibody fragments produced by artificially modifying the antibody light chain or heavy chain variable domain, but are not limited thereto. These antibody fragments can be obtained using proteolytic enzymes (for example, Fab can be obtained by restricting the whole antibody with papain and F(ab')2 fragment can be obtained by cleaving it with pepsin), or produced through recombinant technology, but are not limited thereto.

[0029] In one embodiment, the anti-CD300c antibody of the present invention may be a monoclonal antibody.

[0030] 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, and such monoclonal antibodies exhibit single binding specificity and affinity for a specific epitope.

[0032] Generally, antibodies have a heavy chain and a light chain, and each heavy chain and light chain includes a constant region and a variable region. The variable regions of the light chain and heavy chain include three variable regions called complementarity-determining regions (CDRs) and four structural regions (FRs).

[0033] The above CDRs primarily serve to bind to the epitope of the antigen. The CDRs of each chain are typically named sequentially starting from the N-terminus as CDR1, CDR2, and CDR3, and are also identified by the chain in which a specific CDR is located. The complementarity determining region is situated between regions called the relatively conservative, constant region (FR). Each VH (heavy chain variable region) and VL (light chain variable region) consists of three CDRs and four FRs, arranged in the following order from the amino-terminus to the carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The CDRs of the heavy chain variable region may be referred to as HCDR1, HCDR2, and HCDR3, the CDRs of the light chain variable region as LCDR1, LCDR2, and LCDR3, the FRs of the heavy chain variable region as HFR1, HFR2, HFR3, and HFR4, and the FRs of the light chain variable region as LFR1, LFR2, LFR3, and LFR4.

[0034] In one embodiment, any one or more of the antibody of the present invention and the antigen-binding fragment thereof comprises (i) a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO. 7, SEQ ID NO. 19, SEQ ID NO. 31, SEQ ID NO. 43, SEQ ID NO. 55, SEQ ID NO. 67, SEQ ID NO. 79, SEQ ID NO. 91, SEQ ID NO. 103, SEQ ID NO. 115, SEQ ID NO. 127, SEQ ID NO. 139, SEQ ID NO. 151, SEQ ID NO. 163, SEQ ID NO. 175, SEQ ID NO. 187, SEQ ID NO. 199, SEQ ID NO. 211, SEQ ID NO. 223, SEQ ID NO. 235, SEQ ID NO. 247, SEQ ID NO. 259, SEQ ID NO. 271, SEQ ID NO. 283, and SEQ ID NO. 295; CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO 8, SEQ ID NO 20, SEQ ID NO 32, SEQ ID NO 44, SEQ ID NO 56, SEQ ID NO 68, SEQ ID NO 80, SEQ ID NO 92, SEQ ID NO 104, SEQ ID NO 116, SEQ ID NO 128, SEQ ID NO 140, SEQ ID NO 152, SEQ ID NO 164, SEQ ID NO 176, SEQ ID NO 188, SEQ ID NO 200, SEQ ID NO 212, SEQ ID NO 224, SEQ ID NO 236, SEQ ID NO 248, SEQ ID NO 260, SEQ ID NO 272, SEQ ID NO 284, and SEQ ID NO 296; A heavy chain variable region comprising a CDR3 composed of an amino acid sequence selected from the group consisting of SEQ ID NO. 9, SEQ ID NO. 21, SEQ ID NO. 33, SEQ ID NO. 45, SEQ ID NO. 57, SEQ ID NO. 69, SEQ ID NO. 81, SEQ ID NO. 93, SEQ ID NO. 105, SEQ ID NO. 117, SEQ ID NO. 129, SEQ ID NO. 141, SEQ ID NO. 153, SEQ ID NO. 165, SEQ ID NO. 177, SEQ ID NO. 189, SEQ ID NO. 201, SEQ ID NO. 213, SEQ ID NO. 225, SEQ ID NO. 237, SEQ ID NO. 249, SEQ ID NO. 261, SEQ ID NO. 273, SEQ ID NO. 285, and SEQ ID NO. 297;and (ii) a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO 10, SEQ ID NO 22, SEQ ID NO 34, SEQ ID NO 46, SEQ ID NO 58, SEQ ID NO 70, SEQ ID NO 82, SEQ ID NO 94, SEQ ID NO 106, SEQ ID NO 118, SEQ ID NO 130, SEQ ID NO 142, SEQ ID NO 154, SEQ ID NO 166, SEQ ID NO 178, SEQ ID NO 190, SEQ ID NO 202, SEQ ID NO 214, SEQ ID NO 226, SEQ ID NO 238, SEQ ID NO 250, SEQ ID NO 262, SEQ ID NO 274, SEQ ID NO 286 and SEQ ID NO 298; CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO. 11, SEQ ID NO. 23, SEQ ID NO. 35, SEQ ID NO. 47, SEQ ID NO. 59, SEQ ID NO. 71, SEQ ID NO. 83, SEQ ID NO. 95, SEQ ID NO. 107, SEQ ID NO. 119, SEQ ID NO. 131, SEQ ID NO. 143, SEQ ID NO. 155, SEQ ID NO. 167, SEQ ID NO. 179, SEQ ID NO. 191, SEQ ID NO. 203, SEQ ID NO. 215, SEQ ID NO. 227, SEQ ID NO. 239, SEQ ID NO. 251, SEQ ID NO. 263, SEQ ID NO. 275, SEQ ID NO. 287, and SEQ ID NO. 299; It may include a light chain variable region comprising a CDR3 composed of an amino acid sequence selected from the group consisting of SEQ ID NO. 12, SEQ ID NO. 24, SEQ ID NO. 36, SEQ ID NO. 48, SEQ ID NO. 60, SEQ ID NO. 72, SEQ ID NO. 84, SEQ ID NO. 96, SEQ ID NO. 108, SEQ ID NO. 120, SEQ ID NO. 132, SEQ ID NO. 144, SEQ ID NO. 156, SEQ ID NO. 168, SEQ ID NO. 180, SEQ ID NO. 192, SEQ ID NO. 204, SEQ ID NO. 216, SEQ ID NO. 228, SEQ ID NO. 240, SEQ ID NO. 252, SEQ ID NO. 264, SEQ ID NO. 276, SEQ ID NO. 288, and SEQ ID NO. 300.;

[0035] In one embodiment, in the total amino acid sequence of the heavy chain variable region of the anti-CD300c antibody of the present invention or its antigen-binding fragment, the amino acid sequence from the first amino acid up to CDR1 may be FR1 of the heavy chain variable region; the amino acid sequence from after CDR1 up to CDR2 may be FR2 of the heavy chain variable region; the amino acid sequence from after CDR2 up to CDR3 may be FR3 of the heavy chain variable region; and the sequence from after CDR3 up to the last amino acid of the total amino acid sequence of the heavy chain variable region may be FR4 of the heavy chain variable region. In the total amino acid sequence of the light chain variable region of the anti-CD300c antibody of the present invention or its antigen-binding fragment, the amino acid sequence from the first amino acid up to CDR1 may be FR1 of the light chain variable region; the amino acid sequence from after CDR1 up to CDR2 may be FR2 of the light chain variable region; the amino acid sequence from after CDR2 up to CDR3 may be FR3 of the light chain variable region; and the sequence from after CDR3 up to the last amino acid of the total amino acid sequence of the light chain variable region may be FR4 of the light chain variable region. In addition, the polynucleotides of FR1 to FR4 of the heavy chain variable region and FR1 to FR4 of the light chain variable region can also be defined in the same way as the amino acid sequences. Accordingly, the amino acid sequences of FR1 to FR4 of the heavy chain variable region and FR1 to FR4 of the light chain variable region in the antibody of the present invention and the polynucleotide sequences encoding them can be identified from SEQ ID NOs. 1 to 400.

[0036] In another embodiment, any one or more of the antibody of the present invention and the antigen-binding fragment thereof comprises: (i) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO. 79, SEQ ID NO. 115, and SEQ ID NO. 211; a CDR2 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO. 80, SEQ ID NO. 116, and SEQ ID NO. 212; and a CDR3 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO. 81, SEQ ID NO. 117, and SEQ ID NO. 213; and (ii) a CDR1 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO. 82, SEQ ID NO. 118, and SEQ ID NO. 214; and a CDR2 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO. 83, SEQ ID NO. 119, and SEQ ID NO. 215. It may include a light chain variable region comprising a CDR3 composed of an amino acid sequence selected from the group consisting of SEQ ID NO. 84, SEQ ID NO. 120 and SEQ ID NO. 216.

[0037] As one embodiment of the above-described embodiment, any one or more of the antibody of the present invention and the antigen-binding fragment thereof may be selected from (i) to (iii) below: (i) comprising a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 79, a CDR2 composed of SEQ ID NO. 80, and a CDR3 composed of SEQ ID NO. 81; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 82, a CDR2 composed of SEQ ID NO. 83, and a CDR3 composed of SEQ ID NO. 84; (ii) comprising a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 115, a CDR2 composed of SEQ ID NO. 116, and a CDR3 composed of SEQ ID NO. 117; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 118, a CDR2 composed of SEQ ID NO. 119, and a CDR3 composed of SEQ ID NO. 120. and (iii) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 211, CDR2 composed of SEQ ID NO. 212, and CDR3 composed of SEQ ID NO. 213; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 214, CDR2 composed of SEQ ID NO. 215, and CDR3 composed of SEQ ID NO. 216.

[0038] In one embodiment of the above-described embodiment, any one or more of the antibody of the present invention and the antigen-binding fragment thereof may comprise: (i) a heavy chain variable region comprising a CDR1 composed of an amino acid sequence represented by SEQ ID NO. 79, a CDR2 composed of an amino acid sequence represented by SEQ ID NO. 80, and a CDR3 composed of an amino acid sequence represented by SEQ ID NO. 81; and (ii) a light chain variable region comprising a CDR1 composed of an amino acid sequence represented by SEQ ID NO. 82, a CDR2 composed of an amino acid sequence represented by SEQ ID NO. 83, and a CDR3 composed of an amino acid sequence represented by SEQ ID NO. 84.

[0039] In one embodiment, any one or more of the anti-CD300c antibody of the present invention and the antigen-binding fragment thereof comprises a heavy chain variable region composed of any one of SEQ ID NO 303, SEQ ID NO 307, SEQ ID NO 311, SEQ ID NO 315, SEQ ID NO 319, SEQ ID NO 323, SEQ ID NO 327, SEQ ID NO 331, SEQ ID NO 335, SEQ ID NO 339, SEQ ID NO 343, SEQ ID NO 347, SEQ ID NO 351, SEQ ID NO 355, SEQ ID NO 359, SEQ ID NO 363, SEQ ID NO 367, SEQ ID NO 371, SEQ ID NO 375, SEQ ID NO 379, SEQ ID NO 383, SEQ ID NO 387, SEQ ID NO 391, SEQ ID NO 395, and SEQ ID NO 399; It may include, but is not limited to, a light chain variable region composed of any one of sequence number 304, sequence number 308, sequence number 312, sequence number 316, sequence number 320, sequence number 324, sequence number 328, sequence number 332, sequence number 336, sequence number 340, sequence number 344, sequence number 348, sequence number 352, sequence number 356, sequence number 360, sequence number 364, sequence number 368, sequence number 372, sequence number 376, sequence number 380, sequence number 384, sequence number 388, sequence number 392, sequence number 396, and sequence number 400.

[0040] In one embodiment of the above-described embodiment, any one or more of the anti-CD300c antibody and the antigen-binding fragment thereof may comprise a heavy chain variable region composed of any one of SEQ ID NO. 327, SEQ ID NO. 339, and SEQ ID NO. 371; and a light chain variable region composed of any one of SEQ ID NO. 328, SEQ ID NO. 340, and SEQ ID NO. 372, but are not limited thereto.

[0041] As one embodiment of the above-described embodiment, any one or more of the anti-CD300c antibody and the antigen-binding fragment thereof may include a heavy chain variable region formed by SEQ ID NO. 327 and a light chain variable region formed by SEQ ID NO. 328, but are not limited thereto.

[0042] In one embodiment, any one or more heavy chain variable regions of the anti-CD300c antibody of the present invention and its antigen-binding fragment are encoded by any one of SEQ ID NO. 305, SEQ ID NO. 309, SEQ ID NO. 313, SEQ ID NO. 317, SEQ ID NO. 321, SEQ ID NO. 325, SEQ ID NO. 329, SEQ ID NO. 333, SEQ ID NO. 337, SEQ ID NO. 341, SEQ ID NO. 345, SEQ ID NO. 349, SEQ ID NO. 353, SEQ ID NO. 357, SEQ ID NO. 361, SEQ ID NO. 365, SEQ ID NO. 369, SEQ ID NO. 373, SEQ ID NO. 377, SEQ ID NO. 381, SEQ ID NO. 385, SEQ ID NO. 389, SEQ ID NO. 393, SEQ ID NO. 397, and SEQ ID NO. 341, and any one or more light chain variable regions of the anti-CD300c antibody of the present invention and its antigen-binding fragment are encoded by SEQ ID NO. 306, SEQ ID NO. It may be coded by any one of 310, sequence number 314, sequence number 318, sequence number 322, sequence number 326, sequence number 330, sequence number 334, sequence number 338, sequence number 342, sequence number 346, sequence number 350, sequence number 354, sequence number 358, sequence number 362, sequence number 366, sequence number 370, sequence number 374, sequence number 378, sequence number 382, ​​sequence number 386, sequence number 390, sequence number 394, sequence number 398, and sequence number 402, but is not limited thereto.

[0044] In another embodiment, any one or more of the antibody of the present invention and the antigen-binding fragment thereof may include a heavy chain variable region comprising CDR1 to CDR3, each consisting of an amino acid sequence represented by the following formulas (1) to (3), and a light chain variable region comprising CDR1 to CDR3, each consisting of an amino acid sequence represented by the following formulas (4) to (6):

[0045] FTFX1X2X3X4MX5WVR (1)

[0046] In the above formula,

[0047] X1 = G or S

[0048] X2 = S, R or D

[0049] X3 = N or Y

[0050] X4 = Y, A, G or H

[0051] X5 = S or H

[0052] X1ISX2SGX3X4TYYAX5 (2)

[0053] In the above formula,

[0054] X1 = T or A

[0055] X2 = G or S

[0056] X3 = T or G

[0057] X4 = S or Y

[0058] X5 = D or E

[0059] YCAX1X2X3X4X5X6X7X8X9W (3)

[0060] In the above formula,

[0061] X1 = R or S

[0062] X2 = G or S

[0064] X3 = M, S, Y or I

[0065] X4 = W, Q, G or R

[0066] X5 = G or L

[0067] X6 = M, I or P

[0068] X7 = D, F or L

[0069] X8 = V or D

[0070] X9 = I, Y or not exist

[0071] CX1X2X3X4X5X6X7X8X9X10X11VX12W (4)

[0072] In the above formula,

[0073] X1 = T or S

[0074] X2 = G or R

[0075] X3 = K, N or S

[0076] X4 = H, N or S

[0077] X5 = R, I or G

[0078] X6 = H, G or I

[0079] X7 = T, I or S

[0080] X8 = R, A, K, or does not exist

[0081] X9 = R, S, G, or does not exist

[0082] X10 = N or does not exist

[0083] X11 = Y or does not exist

[0084] X12 = N, H or Q

[0085] X1X2X3X4RPSGVX5 (5)

[0086] In the above formula,

[0087] X1 = L, S, R or E

[0088] X2 = D, K or N

[0089] X3 = S or N

[0090] X4 = E, N, Q or K

[0091] X5 = P or R

[0092] YCX1X2X3X4X5X6X7X8X9X10VF (6)

[0093] In the above formula,

[0094] X1 = Q, A, or S

[0095] X2 = S or A

[0096] X3 = Y or W

[0097] X4 = D or A

[0098] X5 = S, D or G

[0099] X6 = S, N or T

[0100] X7 = S, L, N or K

[0101] X8 = V, S, N or G

[0102] X9 = G, L, V or does not exist

[0103] X10 = P or does not exist.

[0105] In one embodiment, any one or more of the antibody and antigen-binding fragment of the present invention may include one or more of the 25 antibodies shown in Table 1, such as CK1, CK2, CK3, CL4, CL5, CL6, CB201, CL8, CL9, CL10, SK11, SK12, SK13, SK14, SK15, SK16, SK17, SL18, CB301_H3L1_A10, CB301_H3L1_A12, CB301_H3L1_E6, CB301_H3L1_F4, CB301_H3L1_G11, CB301_OPALTL_B5 and CB301_OPALTL_E6.

[0107] antibodies Types of waste paper domain Amino acid sequence (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Sequence number CK1 kappa heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSRYAMTWVR QAPGKGLEWVS SMSGTGGTTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGAYGFDHW GQGTLVTVSS 303 CDR1 FTFSRYAMTWVR 7 CDR2 SMSGTGGTTYYAD 8 CDR3 YCARGAYGFDHW 9 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRASQSIGNYLNW YQQKPGQAPRLLIY DASNLETGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSSAIPYTF GQGTKVEIK 304 CDR1 CRASQSIGNYLNW 10 CDR2 DASNLETGIP 11 CDR3 YCQQSSAIPYTF 12 CK2 kappa heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGTSIYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGGTAFDYW GQGTLVTVSS 307 CDR1 FTFSSYGMHWVR 19 CDR2 AISGSGTSIYYAD 20 CDR3 YCARGGTAFDYW 21 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRASQRSDNYLAW YQQKPGQAPRLLIY DASNRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSYSTPFTF GQGTKVETK 308 CDR1 CRASQRSDNYLAW 22 CDR2 DASNRATGIP 23 CDR3 YCQQSYSTPFTF 24 CK3 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAISWVR QAPGKGLEWVS ATSGSGRATYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARDTWWEGYFDLW GQGTLVTVSS 311 CDR1 FTFSSYAISWVR 31 CDR2 ATSGSGRATYYAD 32 CDR3 YCARDTWWEGYFDLW 33 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CQASHISTHLNW YQQKPGQAPRLLIY GASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQYNTYPPTF GQGTKVEIK 312 CDR1 CQASHISTHLNW 34 CDR2 GASSRATGIP 35 CDR3 YCQQYNTYPPTF 36 CL4 Lambda heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFGSNYMSWVR QAPGKGLEWVS TISGSGTSTYYAD SLKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGMWGMDVW GQGTLVTVSS 315 CDR1 FTFGSNYMSWVR 43 CDR2 TISGSGTSTYYAD 44 CDR3 YCARGMWGMDVW 45 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CTGKHRHTVNW YQLLPGTAPKLLIY LDSERPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCQSYDSSSVVF GGGTKLTVL 316 CDR1 CTGKHRHTVNW 46 CDR2 LDSERPSGVP 47 CDR3 YCQSYDSSSVVF 48 CL5 Lambda heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMHWVR QAPGKGLEWVS SISGGGYGTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSTVWAFDIW GQGTLVTVSS 319 CDR1 FTFSSYAMHWVR 55 CDR2 SISGGGYGTYYAD 56 CDR3 YCARSTVWAFDIW 57 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGSKSVHW YQQLPGTAPKLLIY DVSKRPSERPD RFSGSKSGTSASLAISDLRSEDEADY YCQSFDSSGTWIF GGGTKLTVL 320 CDR1 CSGNNIGSKSVHW 58 CDR2 DVSKRPSERPD 59 CDR3 YCQSFDSSGTWIF 60 CL6 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCAVSGAGRGFFDYW GQGTLVTVSS 323 CDR1 FTFSSYGMHWVR 67 CDR2 AISGSGGSTYYAD 68 CDR3 YCAVSGAGRGFFDYW 69 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGSSSNIGSNYVYW YQQLPGTAPKLLIY EDNKRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYTSSSTVIF GGGTKLTVL 324 CDR1 CSGSSSNIGSNYVYW 70 CDR2 EDNKRPSGVP 71 CDR3 YCSSYTSSSTVIF 72 CB201 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSRYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSSQGIFDIW GQGTLVTVSS 327 CDR1 FTFSRYAMSWVR 79 CDR2 AISGSGGSTYYAD 80 CDR3 YCARSSQGIFDIW 81 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGTRRVHW YQQLPDTAPKLLIY SKNNRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCAAWDDSLSGPVF GGGTKLTVL 328 CDR1 CSGNNIGTRRVHW 82 CDR2 SKNNRPSGVP 83 CDR3 YCAAWDDSLSGPVF 84 CL8 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRNNSKNTLYLQMNSLRAEDTAVY YCARSGRYADLTSG GQGTLVTVSS 331 CDR1 FTFSSYAMSWVR 91 CDR2 AISGSGGSTYYAD 92 CDR3 YCARSGRYADLTSG 93 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGSNSNIGNNYVSW YQQLPDTPPKLLIY DNNKRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYTSSSTVMF GGGTKLTVL 332 CDR1 CSGSNSNIGNNYVSW 94 CDR2 DNNKRPSGVP 95 CDR3 YCSSYTSSSTVMF 96 CL9 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYYWSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARIDVYGFDIW GQGTLVTVSS 335 CDR1 FTFSSYYWSWVR 103 CDR2 AISGSGGSTYYAD 104 CDR3 YCARIDVYGFDIW 105 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGSTSNIGTNYVYW YQQLPGTAPKLLIY DNNNRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCQTWDSSTDVVF GGGTKLTVL 336 CDR1 CSGSTSNIGTNYVYW 106 CDR2 DNNNRPSGVP 107 CDR3 YCQTWDSSTDVVF 108 CL10 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCASGYGLMDVW GQGTLVTVSS 339 CDR1 FTFSSYGMHWVR 115 CDR2 AISGSGGSTYYAD 116 CDR3 YCASGYGLMDVW 117 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CTRSSGIIASNYVQW YQQLPGTAPKLLIY RNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYAGNNNLVF GGGTKLTVL 340 CDR1 CTRSSGIIASNYVQW 118 CDR2 RNNQRPSGVP 119 CDR3 YCSSYAGNNNLVF 120 SK11 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSTYGMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGLSGLDYW GQGTLVTVSS 343 CDR1 FTFSTYGMHWVR 127 CDR2 AISGSGGSTYYAD 128 CDR3 YCARGLSGLDYW 129 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRSSQGITNYLAW YQQKPGQAPRLLIY DASNRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSYSTPLTF GQGTKVEIK 344 CDR1 CRSSQGITNYLAW 130 CDR2 DASNRATGIP 131 CDR3 YCQQSYSTPLTF 132 SK12 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMHWVR QAPGKGLEWVS AISGSGGDTYHAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCTRGLSGFDYW GQGTLVTVSS 347 CDR1 FTFSSYAMHWVR 139 CDR2 AISGSGGDTYHAD 140 CDR3 YCTRGLSGFDYW 141 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRASQSISSYLNW YQQKPGQAPRLLIY DASNRAPGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSYSIPITF GQGTKVEIKFSD 348 CDR1 CRASQSISSYLNW 142 CDR2 DASNRAPGIP 143 CDR3 YCQQSYSIPITF 144 SK13 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSDYAMSWVR QAPGKGLEWVS SISSSSSYIYYTD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGGYGFDYW GQGTLVTVSS 351 CDR1 FTFSDYAMSWVR 151 CDR2 SISSSSSYIYYTD 152 CDR3 YCARGGYGFDYW 153 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRASQSISSYLNW YQQKPGQAPRLLIY SASSRPQGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQYDDLPFTF GQGTKVEIK 352 CDR1 CRASQSISSYLNW 154 CDR2 SASSRPQGIP 155 CDR3 YCQQYDDLPFTF 156 SK14 kappa heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSNFAIAWVR QAPGKGLEWVS AISGRGTSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGVSGFDSW GQGTLVTVSS 355 CDR1 FTFSNFAIAWVR 163 CDR2 AISGRGTSTYYAD 164 CDR3 YCARGVSGFDSW 165 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRASQSISSHLAW YQQKPGQAPRLLIY DTSNRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSYSTPFTF GQGTKVEIK 356 CDR1 CRASQSISSHLAW 166 CDR2 DTSNRATGIP 167 CDR3 YCQQSYSTPFTF 168 SK15 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMHWVR QAPGKGLEWVS AINGSGGSTYYAD SVKGRFTISRDNSKNTLYLQTNSLRAEDTAVY YCARGLQGFDYW GQGTLVTVSS 359 CDR1 FTFSSYAMHWVR 175 CDR2 AINGSGGSTYYAD 176 CDR3 YCARGLQGFDYW 177 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CQASQDITNYLNW YQQKPGQAPRLLIY DASSLETGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSYSTPITF GQGTKVEIK 360 CDR1 CQASQDITNYLNW 178 CDR2 DASSLETGIP 179 CDR3 YCQQSYSTPITF 180 SK16 kappa heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMSWVR QAPGKGLEWVS AINGSGGSTLYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGVSGFDSW GQGTLVTVSS 363 CDR1 FTFSSYAMSWVR 187 CDR2 AINGSGGSTLYAD 188 CDR3 YCARGVSGFDSW 189 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CRISQSISSYLNW YQQKPGQAPRLLIY DASLRATGIP DRFSGSGSGTDFTLTISRLEPEDFAV YYCQQSYKTPITF GQGTKVEIK 364 CDR1 CRISQSISSYLNW 190 CDR2 DASLRATGIP 191 CDR3 YYCQQSYKTPITF 192 SK17 kappa heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYYWSWVR QAPGKGLEWVS TITGSGGSTDYAN SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCATGGGIFDYW GQGTLVTVSS 367 CDR1 FTFSSYYWSWVR 199 CDR2 TITGSGGSTDYAN 200 CDR3 YCATGGGIFDYW 201 Light chain variable region entire EIVLTQSPGTLSLSPGERATLS CQASQTISNYLNW YQQKPGQAPRLLIY DASNRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVY YCQQYNSYPPSF GQGTKVEIK 368 CDR1 CQASQTISNYLNW 202 CDR2 DASNRATGIP 203 CDR3 YCQQYNSYPPSF 204 SL18 Lambda heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSDYHMHWVR QAPGKGLEWVS TISSSGGYTYYAE SVKSRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSIRLPLDYW GQGTLVTVSS 371 CDR1 FTFSDYHMHWVR 211 CDR2 TISSSGGYTYYAE 212 CDR3 YCARSIRLPLDYW 213 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGSKGVHW YQQLPGTAPKLLIY EDSKRPSGVR DRFSGSKSGTSASLAISGLRSEDEADY YCQSYDSTKGVVF GGGTKLTVL 372 CDR1 CSGNNIGSKGVHW 214 CDR2 EDSKRPSGVR 215 CDR3 YCQSYDSTKGVVF 216 CB301_H3L1_A10 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQSGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCVRGYGAMDVW GQGTLVTVSS 375 CDR1 FTFSSYGMHWVR 223 CDR2 AISGSGGSTYYAD 224 CDR3 YCVRGYGAMDVW 225 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CTRSSGSIASNYVQW YQQLPGTAPKLLIY RNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYTTSSTLVF GGGTKLTVL 376 CDR1 CTRSSGSIASNYVQW 226 CDR2 RNNQRPSGVP 227 CDR3 YCSSYTTSSTLVF 228 CB301_H3L1_A12 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAKDTAVY YCASGYGLMDVW GQGTLVTVSS 379 CDR1 FTFSSYAMHWVR 235 CDR2 AISGSGGSTYYAD 236 CDR3 YCASGYGLMDVW 237 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CTGTSSDVGNYNLVSW YQQLPGTAPKLLIY SNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYTGSNALLF GGGTKLTVL 380 CDR1 CTGTSSDVGNYNLVSW 238 CDR2 SNNQRPSGVP 239 CDR3 YCSSYTGSNALLF 240 CB301_H3L1_E6 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARWHYSFDYW GQGTLVTVSS 383 CDR1 FTFSSYAMSWVR 247 CDR2 AISGSGGSTYYAD 248 CDR3 YCARWHYSFDYW 249 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CRGNNIGSKRVHW YQQLPGTAPKLLIY SYNHRPSGVP DRFSGSKSGTSASLAITGLRSEDEADY YCNTWDDSLEGPVF GGGTKLTVL 384 CDR1 CRGNNIGSKRVHW 250 CDR2 SYNHRPSGVP 251 CDR3 YCNTWDDSLEGPVF 252 CB301_H3L1_F4 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSGYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSPSGLFDYW GQGTLVTVSS 387 CDR1 FTFSGYAMSWVR 259 CDR2 AISGSGGSTYYAD 260 CDR3 YCARSPSGLFDYW 261 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CGGNNIGSKRVHW YQQLPGTAPKLLIY NTSNKHSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYLQQHSLF GGGTKLTVL 388 CDR1 CGGNNIGSKRVHW 262 CDR2 NTSNKHSGVP 263 CDR3 YCSSYLQQHSLF 264 CB301_H3L1_G11 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSSSASSCAASG FTFSSYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCTRFVGAIGAFDYW GQGTLVTVSS 391 CDR1 FTFSSYAMSWVR 271 CDR2 AISGSGGSTYYAD 272 CDR3 YCTRFVGAIGAFDYW 273 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGSRSVHW YQQLPGTAPKLLIY RNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCAAWDDSLSGPVF GGGTKLTVL 392 CDR1 CSGNNIGSRSVHW 274 CDR2 RNNQRPSGVP 275 CDR3 YCAAWDDSLSGPVF 276 CB301_OPALTL_B5 OPALTL heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSHYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARGWDSPTLTYFDSW GQGTLVTVSS 395 CDR1 FTFSHYAMSWVR 283 CDR2 AISGSGGSTYYAD 284 CDR3 YCARGWDSPTLTYFDSW 285 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGTSSNIGNNDVSW YQQLPGTAPKLLIY QDTKRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCAAWDDSLSGPVF GGGTKLTVL 396 CDR1 CSGTSSNIGNNDVSW 286 CDR2 QDTKRPSGVP 287 CDR3 YCAAWDDSLSGPVF 288 CB301_OPALTL_E6 OPALTL heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGGYTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARWHYSFDYW GQGTLVTVSS 399 CDR1 FTFSSYGMHWVR 295 CDR2 AISGSGGYTYYAD 296 CDR3 YCARWHYSFDYW 297 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGSSSNIGNNYVSW YQQLPGTAPKLLIY RNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCQSYDNSNVLF GGGTKLTVL 400 CDR1 CSGSSSNIGNNYVSW 298 CDR2 RNNQRPSGVP 299 CDR3 YCQSYDNSNVLF 300

[0108] In the present invention, the variable region CDR was determined by a conventional method according to a system devised by Kabat et al. (see reference [Kabat et al., Sequences of Proteins of Immunological Interest (5th), National Institutes of Health, Bethesda, MD. (1991)]). Although the CDR numbering used in the present invention utilized the Kabat method, antibodies containing CDRs determined by other methods, such as the IMGT method, the Chothia method, and the AbM method, are also included within the scope of the present invention.

[0109] When the antibody of the present invention includes a constant region, it may include a constant region derived from IgG, IgA, IgD, IgE, IgM, or a combination thereof or a hybrid thereof.

[0110] In the present invention, the term “combination” means that when forming a dimer or a multimer, a polypeptide encoding a single-chain immunoglobulin constant region of the same origin forms a combination with a single-chain polypeptide of a different origin. For example, a dimer or a multimer can be formed from two or more constant regions selected from the group consisting of the constant regions of IgG, IgA, IgD, IgE, and IgM.

[0111] In the present invention, the term "hybrid" means that there are sequences corresponding to two or more immunoglobulin heavy chain constant regions of different origin within a short-chain immunoglobulin heavy chain constant region, and, for example, a hybrid of a domain consisting of one to four domains selected from the group consisting of CH1, CH2, CH3, and CH4 of IgG, IgA, IgD, IgE, and IgM is possible.

[0112] In addition, in the present invention, the origin of the variable region and the constant region of the antibody may be the same or different, and the origin of the variable region and the constant region excluding the CDR may be the same or different.

[0114] As one embodiment of the above-described embodiment, any one or more of the anti-CD300c antibody and antigen-binding fragment may include a sequence having sequence identity of 80% or more, 85% or more, 90% or more, more preferably 95% or more, and most preferably 98% or more compared to the CDR sequence or heavy chain variable region and light chain variable region sequence presented in Table 1.

[0115] As an exemplary embodiment, amino acid sequence variants of the antibody of the present invention may be considered. Variants produced to the extent generally practiced in the art to maintain or enhance the function of the anti-CD300c antibody and its antigen-binding fragment of the present invention, provided that the anti-CD300c antibody and its antigen-binding fragment of the present invention possess characteristics identical to or equivalent to those of the present invention, may also be included in the present invention. For example, this may involve improving the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of the antibody may be produced by introducing appropriate modifications into the nucleotide sequence encoding the molecule or by peptide synthesis. Such modifications include, for example, the deletion of residues from the amino acid sequence of the antibody and / or the insertion of residues into such amino acid sequence and / or the substitution of residues within such amino acid sequence. Any combination of deletions, insertions, and substitutions may be performed to arrive at a final construct, but the final construct must possess desired properties, for example, antigen-binding properties. The sites of interest for substitution mutagenicity include the heavy chain variable region (HVR) and the backbone region (FR). Conservative substitutions are provided in Table 2 under the item “Desired substitutions” and are further described below in relation to amino acid side chain classes (1) to (6). Amino acid substitutions can be introduced into products screened for molecules of interest and desired activities, e.g., maintained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0117] Original residual Exemplary Substitution Desirable substitution Ala(A) Val; Leu; Ile Val Arg(R) Lys; Gln; Asn Lys Asn(N) Gln; His; Asp; Lys; Arg Gln Asp(D) Glu; Asn Glu Cys(C) Ser; Ala Ser Gln(Q) Asn; Glu Asn Glu(E) Asp; Gln Asp Gly(G) Ala Ala His(H) Asn; Gln; Lys; Arg Arg Ile(I) Leu; Val; Met; Ala; Phe; Norleucine Leu Leu(L) Norleucine; Ile; Val; Met; Ala; Phe Ile Lys(K) Arg; Gln; Asn Arg Met(M) Leu; Phe; Ile Ile Phe(F) Trp; Leu; Val; Ile; Ala; Tyr Tyr Pro(P) Ala Ala Ser(S) Thr Thr Thr(T) Val; Ser Ser Trp(W) Tyr; Phe Tyr Tyr(Y) Trp; Phe; Thr; Ser Phe Val(V) Ile; Leu; Met; Phe; Ala; Norleucine Leu

[0118] Amino acids can be grouped as follows according to their typical side chain properties:

[0119] (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile;

[0120] (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

[0121] (3) Acids: Asp, Glu;

[0122] (4) Basics: His, Lys, Arg;

[0123] (5) Residues affecting chain orientation: Gly, Pro;

[0124] (6) Aromatic: Trp, Tyr, Phe.

[0125] Non-conservative substitution involves exchanging a member of one of these classes for another.

[0126] In the present invention, the term "amino acid sequence variant" comprises a substantial variant in which amino acid substitutions exist at one or more hypervariable region residues of a parent antibody-binding molecule (e.g., humanized or human antibody). Generally, variants selected and generated for further study will have modifications, such as improvements (e.g., increased affinity, decreased immunogenicity) of specific biological properties compared to the parent antibody-binding molecule, or will substantially retain specific biological properties of the parent antigen-binding molecule. An exemplary substitution variant is an affinity-maturing antibody, which can be conveniently generated, for example, using phage display-based affinity maturation techniques known in the art. Briefly, one or more HVR (Hyper-variable region) residues are mutated, and variant antigen-binding molecules are displayed on phages to screen for specific biological activities (e.g., binding affinity). In a specific embodiment, substitutions, insertions, or deletions may occur within one or more HVRs, provided that such changes do not substantially reduce the ability of the antigen-binding molecule to bind to the antigen. For example, conservative modifications that do not substantially reduce binding affinity (e.g., conservative substitutions as provided herein) may be made in HVR. Amino acid sequence insertions may include the insertion of a single or multiple amino acid residues into the sequence, as well as amino-terminal and / or carboxyl-terminal fusions ranging in length from one residue to a polypeptide containing one hundred or more residues. Examples of terminal insertions include antibodies having an N-terminal methionyl residue. Other insertion variants of the molecule may include fusions to the N-terminus or C-terminus of the polypeptide to increase the serum half-life of the antibody. Additionally, other insertion variants of the molecule may include fusions to the N-terminus or C-terminus of the polypeptide to facilitate passage across the blood-brain barrier (BBB).

[0127] In the present invention, the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CK1 are each SEQ ID NOs 1 to 3; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CK1 are each SEQ ID NOs 4 to 6; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CK1 are each SEQ ID NOs 7 to 9; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CK1 are each SEQ ID NOs 10 to 12; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CK2 are each SEQ ID NOs 13 to 15; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CK2 are each SEQ ID NOs 16 to 18. The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody CK2 are each SEQ NOs 19 to 21; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody CK2 are each SEQ NOs 22 to 24; the polynucleotide sequences encoding the heavy chain variable region CDR1 to CDR3 of antibody CK3 are each SEQ NOs 25 to 27; the polynucleotide sequences encoding the light chain variable region CDR1 to CDR3 of antibody CK3 are each SEQ NOs 28 to 30; the amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody CK3 are each SEQ NOs 31 to 33; and the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody CK3 are each SEQ NOs 34 to 36; The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL4 are each SEQ ID NOs 37 to 39; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL4 are each SEQ ID NOs 40 to 42; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CL4 are each SEQ ID NOs 43 to 45;The amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CL4 are each SEQ ID NOs 46 to 48; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL5 are each SEQ ID NOs 49 to 51; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL5 are each SEQ ID NOs 52 to 54; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CL5 are each SEQ ID NOs 55 to 57; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL5 are each SEQ ID NOs 58 to 60; and the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ ID NOs 61 to 63. The polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 64 to 66; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 67 to 69; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 70 to 72; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 61 to 63; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 64 to 66; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ NOs 67 to 69; The amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CL6 are each SEQ ID NOs 70 to 72; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB201 are each SEQ ID NOs 73 to 75; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB201 are each SEQ ID NOs 76 to 78.The amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CB201 are each SEQ NOs 79 to 81; the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CB201 are each SEQ NOs 82 to 84; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL8 are each SEQ NOs 85 to 87; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL8 are each SEQ NOs 88 to 90; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CL8 are each SEQ NOs 91 to 93; and the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CL8 are each SEQ NOs 94 to 96. The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL9 are each SEQ NOs 97 to 99; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL9 are each SEQ NOs 100 to 102; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL9 are each SEQ NOs 103 to 105; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL9 are each SEQ NOs 106 to 108; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CL10 are each SEQ NOs 109 to 111; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CL10 are each SEQ NOs 112 to 114; The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody CL10 are each SEQ ID NOs 115 to 117; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody CL10 are each SEQ ID NOs 118 to 120;The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK11 are each SEQ NOs 121 to 123; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK11 are each SEQ NOs 124 to 126; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK11 are each SEQ NOs 127 to 129; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK11 are each SEQ NOs 130 to 132; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK12 are each SEQ NOs 133 to 135; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK12 are each SEQ NOs 136 to 138. The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody SK12 are each SEQ NOs 139 to 141; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody SK12 are each SEQ NOs 142 to 144; the polynucleotide sequences encoding the heavy chain variable region CDR1 to CDR3 of antibody SK13 are each SEQ NOs 145 to 147; the polynucleotide sequences encoding the light chain variable region CDR1 to CDR3 of antibody SK13 are each SEQ NOs 148 to 150; the amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody SK13 are each SEQ NOs 151 to 153; and the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody SK13 are each SEQ NOs 154 to 156. The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK14 are each SEQ ID NOs 157 to 159; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK14 are each SEQ ID NOs 160 to 162;The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody SK14 are each SEQ NOs 163 to 165; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody SK14 are each SEQ NOs 166 to 168; the polynucleotide sequences encoding the heavy chain variable region CDR1 to CDR3 of antibody SK15 are each SEQ NOs 169 to 171; the polynucleotide sequences encoding the light chain variable region CDR1 to CDR3 of antibody SK15 are each SEQ NOs 172 to 174; the amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody SK15 are each SEQ NOs 175 to 177; and the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody SK15 are each SEQ NOs 178 to 180. The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK16 are each SEQ NOs 181 to 183; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK16 are each SEQ NOs 184 to 186; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK16 are each SEQ NOs 187 to 189; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK16 are each SEQ NOs 190 to 192; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SK17 are each SEQ NOs 193 to 195; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SK17 are each SEQ NOs 196 to 198. The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody SK17 are each SEQ ID NOs 199 to 201; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody SK17 are each SEQ ID NOs 202 to 204;The polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody SL18 are each SEQ ID NOs 205 to 207; the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody SL18 are each SEQ ID NOs 208 to 210; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody SL18 are each SEQ ID NOs 211 to 213; the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody SL18 are each SEQ ID NOs 214 to 216; and the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A10 are each SEQ ID NOs 217 to 219. The polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A10 are each SEQ ID NOs 220 to 222; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A10 are each SEQ ID NOs 223 to 225; the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A10 are each SEQ ID NOs 226 to 228; and the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A12 are each SEQ ID NOs 229 to 231. The polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A12 are each SEQ ID NOs 232 to 234; the amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A12 are each SEQ ID NOs 235 to 237; the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_A12 are each SEQ ID NOs 238 to 240; and the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_E6 are each SEQ ID NOs 241 to 243.The polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_E6 are each SEQ ID NOs 244 to 246; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_E6 are each SEQ ID NOs 247 to 249; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_E6 are each SEQ ID NOs 250 to 252; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CCB301_H3L1_F4 are each SEQ ID NOs 253 to 255; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_F4 are each SEQ ID NOs 256 to 258. The amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody CB301_H3L1_F4 are each SEQ ID NOs 259 to 261; the amino acid sequences of the light chain variable region CDR1 to CDR3 of antibody CB301_H3L1_F4 are each SEQ ID NOs 262 to 264; the polynucleotide sequences encoding the heavy chain variable region CDR1 to CDR3 of antibody CB301_H3L1_G11 are each SEQ ID NOs 265 to 267; the polynucleotide sequences encoding the light chain variable region CDR1 to CDR3 of antibody CB301_H3L1_G11 are each SEQ ID NOs 268 to 270; and the amino acid sequences of the heavy chain variable region CDR1 to CDR3 of antibody CB301_H3L1_G11 are each SEQ ID NOs 271 to 273. The amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CB301_H3L1_G11 are SEQ ID NOs 274 to 276, respectively; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_B5 are SEQ ID NOs 277 to 279, respectively;The polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_B5 are each SEQ ID NOs 280 to 282; the amino acid sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_B5 are each SEQ ID NOs 283 to 285; the amino acid sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_B5 are each SEQ ID NOs 286 to 288; the polynucleotide sequences encoding the heavy chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_E6 are each SEQ ID NOs 289 to 291; and the polynucleotide sequences encoding the light chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_E6 are each SEQ ID NOs 292 to 294. The amino acid sequences of the heavy chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_E6 are shown in SEQ ID NOs 295 to 297, respectively; and the amino acid sequences of the light chain variable regions CDR1 to CDR3 of antibody CB301_OPALTL_E6 are shown in SEQ ID NOs 298 to 300, respectively.

[0128] In addition, the polynucleotide sequence encoding the heavy chain variable region of antibody CK1, the polynucleotide sequence encoding the light chain variable region of antibody CK1, the amino acid sequence of the heavy chain variable region of antibody CK1, and the amino acid sequence of the light chain variable region of antibody CK1 are each SEQ ID NOs 301 to 304; the polynucleotide sequence encoding the heavy chain variable region of antibody CK2, the polynucleotide sequence encoding the light chain variable region of antibody CK2, the amino acid sequence of the heavy chain variable region of antibody CK2, and the amino acid sequence of the light chain variable region of antibody CK2 are each SEQ ID NOs 305 to 308; the polynucleotide sequence encoding the heavy chain variable region of antibody CK3, the polynucleotide sequence encoding the light chain variable region of antibody CK3, the amino acid sequence of the heavy chain variable region of antibody CK3, and the amino acid sequence of the light chain variable region of antibody CK3 are each SEQ ID NOs 309 to 312; The polynucleotide sequence encoding the heavy chain variable region of antibody CL4, the polynucleotide sequence encoding the light chain variable region of antibody CL4, the amino acid sequence of the heavy chain variable region of antibody CL4, and the amino acid sequence of the light chain variable region of antibody CL4 are each SEQ ID NOs 313 to 316; the polynucleotide sequence encoding the heavy chain variable region of antibody CL5, the polynucleotide sequence encoding the light chain variable region of antibody CL5, the amino acid sequence of the heavy chain variable region of antibody CL5, and the amino acid sequence of the light chain variable region of antibody CL5 are each SEQ ID NOs 317 to 320; the polynucleotide sequence encoding the heavy chain variable region of antibody CL6, the polynucleotide sequence encoding the light chain variable region of antibody CL6, the amino acid sequence of the heavy chain variable region of antibody CL6, and the amino acid sequence of the light chain variable region of antibody CL6 are each SEQ ID NOs 321 to 324; The polynucleotide sequence encoding the heavy chain variable region of antibody CB201, the polynucleotide sequence encoding the light chain variable region of antibody CB201, the amino acid sequence of the heavy chain variable region of antibody CB201, and the amino acid sequence of the light chain variable region of antibody CB201 are each SEQ ID NOs 325 to 328;The polynucleotide sequence encoding the heavy chain variable region of antibody CL8, the polynucleotide sequence encoding the light chain variable region of antibody CL8, the amino acid sequence of the heavy chain variable region of antibody CL8, and the amino acid sequence of the light chain variable region of antibody CL8 are each SEQ ID NOs 329 to 332; the polynucleotide sequence encoding the heavy chain variable region of antibody CL9, the polynucleotide sequence encoding the light chain variable region of antibody CL9, the amino acid sequence of the heavy chain variable region of antibody CL9, and the amino acid sequence of the light chain variable region of antibody CL9 are each SEQ ID NOs 333 to 336; the polynucleotide sequence encoding the heavy chain variable region of antibody CL10, the polynucleotide sequence encoding the light chain variable region of antibody CL10, the amino acid sequence of the heavy chain variable region of antibody CL10, and the amino acid sequence of the light chain variable region of antibody CL10 are each SEQ ID NOs 337 to 340; The polynucleotide sequence encoding the heavy chain variable region of antibody SK11, the polynucleotide sequence encoding the light chain variable region of antibody SK11, the amino acid sequence of the heavy chain variable region of antibody SK11, and the amino acid sequence of the light chain variable region of antibody SK11 are each SEQ ID NOs 341 to 344; the polynucleotide sequence encoding the heavy chain variable region of antibody SK12, the polynucleotide sequence encoding the light chain variable region of antibody SK12, the amino acid sequence of the heavy chain variable region of antibody SK12, and the amino acid sequence of the light chain variable region of antibody SK12 are each SEQ ID NOs 345 to 348; the polynucleotide sequence encoding the heavy chain variable region of antibody SK13, the polynucleotide sequence encoding the light chain variable region of antibody SK13, the amino acid sequence of the heavy chain variable region of antibody SK13, and the amino acid sequence of the light chain variable region of antibody SK13 are each SEQ ID NOs 349 to 352; The polynucleotide sequence encoding the heavy chain variable region of antibody SK14, the polynucleotide sequence encoding the light chain variable region of antibody SK14, the amino acid sequence of the heavy chain variable region of antibody SK14, and the amino acid sequence of the light chain variable region of antibody SK14 are each SEQ ID NOs 353 to 356;The polynucleotide sequence encoding the heavy chain variable region of antibody SK15, the polynucleotide sequence encoding the light chain variable region of antibody SK15, the amino acid sequence of the heavy chain variable region of antibody SK15, and the amino acid sequence of the light chain variable region of antibody SK15 are each SEQ ID NOs 357 to 360; the polynucleotide sequence encoding the heavy chain variable region of antibody SK16, the polynucleotide sequence encoding the light chain variable region of antibody SK16, the amino acid sequence of the heavy chain variable region of antibody SK16, and the amino acid sequence of the light chain variable region of antibody SK16 are each SEQ ID NOs 361 to 364; the polynucleotide sequence encoding the heavy chain variable region of antibody SK17, the polynucleotide sequence encoding the light chain variable region of antibody SK17, the amino acid sequence of the heavy chain variable region of antibody SK17, and the amino acid sequence of the light chain variable region of antibody SK17 are each SEQ ID NOs 365 to 368; The polynucleotide sequence encoding the heavy chain variable region of antibody SL18, the polynucleotide sequence encoding the light chain variable region of antibody SL18, the amino acid sequence of the heavy chain variable region of antibody SL18, and the amino acid sequence of the light chain variable region of antibody SL18 are each SEQ ID NOs 369 to 372; the polynucleotide sequence encoding the heavy chain variable region of antibody CB301_H3L1_A10, the polynucleotide sequence encoding the light chain variable region of antibody CB301_H3L1_A10, the amino acid sequence of the heavy chain variable region of antibody CB301_H3L1_A10, and the amino acid sequence of the light chain variable region of antibody CB301_H3L1_A10 are each SEQ ID NOs 373 to 376; The polynucleotide sequence encoding the heavy chain variable region of antibody CB301_H3L1_A12, the polynucleotide sequence encoding the light chain variable region of antibody CB301_H3L1_A12, the amino acid sequence of the heavy chain variable region of antibody CB301_H3L1_A12, and the amino acid sequence of the light chain variable region of antibody CB301_H3L1_A12 are each SEQ ID NOs 377 to 380;The polynucleotide sequence encoding the heavy chain variable region of antibody CB301_H3L1_E6, the polynucleotide sequence encoding the light chain variable region of antibody CB301_H3L1_E6, the amino acid sequence of the heavy chain variable region of antibody CB301_H3L1_E6, and the amino acid sequence of the light chain variable region of antibody CB301_H3L1_E6 are each SEQ ID NOs 381 to 384; the polynucleotide sequence encoding the heavy chain variable region of antibody CB301_H3L1_F4, the polynucleotide sequence encoding the light chain variable region of antibody CB301_H3L1_F4, the amino acid sequence of the heavy chain variable region of antibody CB301_H3L1_F4, and the amino acid sequence of the light chain variable region of antibody CB301_H3L1_F4 are each SEQ ID NOs 385 to 388; The polynucleotide sequence encoding the heavy chain variable region of antibody CB301_H3L1_G11, the polynucleotide sequence encoding the light chain variable region of antibody CB301_H3L1_G11, the amino acid sequence of the heavy chain variable region of antibody CB301_H3L1_G11, and the amino acid sequence of the light chain variable region of antibody CB301_H3L1_G11 are each SEQ ID NOs 389 to 392; the polynucleotide sequence encoding the heavy chain variable region of antibody CB301_OPALTL_B5, the polynucleotide sequence encoding the light chain variable region of antibody CB301_OPALTL_B5, the amino acid sequence of the heavy chain variable region of antibody CB301_OPALTL_B5, and the amino acid sequence of the light chain variable region of antibody CB301_OPALTL_B5 are each SEQ ID NOs 393 to 396; The polynucleotide sequence encoding the heavy chain variable region of antibody CB301_OPALTL_E6, the polynucleotide sequence encoding the light chain variable region of antibody CB301_OPALTL_E6, the amino acid sequence of the heavy chain variable region of antibody CB301_OPALTL_E6, and the amino acid sequence of the light chain variable region of antibody CB301_OPALTL_E6 are each SEQ ID NOs. 397 to 400; and the amino acid sequence of CD300C ECD and the polynucleotide sequence encoding it are each shown in SEQ ID NOs. 401 and 402.;

[0130] One or more of the antibodies of the present invention and their antigen-binding fragments may induce the promotion of differentiation into MDM (Monocyte-derived macrophage) or M1 macrophage through engagement with CD300c, but are not limited thereto.

[0131] In one embodiment of the above-described embodiment, the anti-CD00c antibody and its antigen-binding fragment can specifically bind to the extracellular domain of the CD300c protein. The extracellular domain of the CD300c may be the extracellular domain of the human CD300c protein and may include the amino acid sequence represented by SEQ ID NO. 402.

[0132] In one embodiment, the antibody and / or antigen-binding fragment thereof of the present invention has the ability to restore or improve cognitive function of a subject having or at risk of a degenerative brain disease, said cognitive function may include cognitive ability, memory ability, language ability, visuospatial ability, orientation, executive ability, or a combination thereof.

[0134] In one embodiment, the antibody of the present invention may have the ability to promote differentiation from monocytes into MDM (Monocyte-derived macrophage) or M1 macrophage, and may have said promoting ability increased compared to general neurodegenerative disease treatments and immunotherapies. As a result, it is expected that it may exhibit a significant preventive or therapeutic effect against neurodegenerative disease even in patients with neurodegenerative disease who have not been treated by general neurodegenerative disease treatments and immunotherapies.

[0136] In the present invention, the term "degenerative brain disease" refers to a brain disease caused by damage to nerve cells and may be accompanied by cognitive impairment. The cognitive impairment may include a decline in cognitive ability, memory impairment, language impairment, decline in visuospatial ability, orientation impairment, executive function impairment, or a combination thereof.

[0137] In one embodiment, the degenerative brain disease may include, but is not limited to, one or more selected from the group consisting of dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, cerebral amyloid angiopathy, amyloid stroke, Dutch amyloidosis, tauopathy, mild cognitive impairment, corticobasal degeneration, posterior cortical atrophy, primary progressive aphasia, progressive supranuclear palsy, corticobasal deteneration (CBD), amyotrophic lateral sclerosis, Creutzfeldt-Jakob disease, amnesia, learning disability, and memory impairment.

[0138] The above dementia refers to a decline or progressive decline in cognitive function caused by brain damage or disease, and generally may include disorientation, impairment of memory, judgment and intelligence, unstable emotions, or a combination thereof. The above dementia may include, but is not limited to, one or more selected from the group including, Alzheimer's disease dementia, Huntington's disease dementia, Parkinson's disease dementia, cerebrovascular dementia, neuroinflammatory dementia, cerebral infarction dementia, senile dementia, Lewy body dementia (Dimentia with Lewy bodies, DLB), Multi-Infarct Dementia (MID), Frontotemporal lobar degeneration (FTLD), Creutzfeldt-Jakob disease dementia, Pick's disease dementia, corticobasal degeneration dementia, dementia due to normal pressure hydrocephalus, and dementia due to head trauma.

[0139] As an example of implementation, a degenerative brain disease could be Alzheimer's disease.

[0140] In one embodiment of the present invention, through a Y-maze test using a 5xFAD mouse with a degenerative brain disease, it was confirmed that the change behavioral ability reduced by the induction of the degenerative brain disease was statistically significantly restored by the administration of an anti-CD300c antibody.

[0141] In addition, in another embodiment of the present invention, it was confirmed through the Morris water maze test that the cognitive ability and memory of mice reduced by the induction of brain disease were statistically significantly restored by the administration of an anti-CD300c antibody.

[0142] Furthermore, in another embodiment of the present invention, it was confirmed that the anti-CD300c antibody of the present invention possesses the ability to promote differentiation from monocytes into MDM (Monocyte-derived macrophage) or M1 macrophages. The neurodegenerative diseases of the present invention may be prevented or treated by promoting differentiation from monocytes into MDM or M1 macrophages, and these results suggest that the anti-CD300c antibody of the present invention and / or its antigen-binding fragment may also treat various neurodegenerative diseases (e.g., Huntington's disease and Parkinson's disease, etc.) that are treated by promoting differentiation into MDM or M1 macrophages.

[0144] The composition of the present invention prevents or treats degenerative brain diseases, wherein any one or more of the antibodies and antigen-binding fragments included therein may restore or improve the cognitive function of a subject having a degenerative brain disease or at risk thereof.

[0145] In the present invention, the term "treatment" generally means obtaining desired pharmacological and / or physiological effects and may include any act in which the symptoms of a degenerative brain disease are improved, delayed, or beneficially altered by the administration of the composition of the present invention. Furthermore, an act in which the current state is maintained without the worsening of symptoms due to the nature of the brain disease may also be included in the treatment. These effects are therapeutic in that they partially or completely cure the disease and / or the side effects caused by such disease. Desirable therapeutic effects include, but are not limited to, prevention of the onset or recurrence of the disease, improvement of symptoms, reduction of any direct or indirect pathological consequences of the disease, reduction of the rate of disease progression, improvement or alleviation of the disease state, and remission or improved prognosis.

[0146] In the present invention, the term "prevention" relates to preventive treatment, that is, obtaining an effect to prevent rather than treat a disease. Such prevention means obtaining a desired preventive pharmacological effect and / or physiological effect in the sense of partially or completely preventing a disease or its symptoms.

[0148] The above pharmaceutical composition may further include a pharmaceutically acceptable carrier.

[0149] In the present invention, the term "pharmaceuticalally acceptable carrier" refers to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, provided that they are sterile and biocompatible. Additionally, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, as well as pills, capsules, granules, or tablets.

[0150] The above pharmaceutical composition may be in various dosage forms for oral or parenteral administration. When formulated, it is prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and these solid dosage forms are prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin, with one or more compounds. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid dosage forms for oral administration include suspensions, liquids, emulsions, and syrups, and may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Preparations for parenteral administration include, for example, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrogol, Tween 61, cocoa paste, laurin paste, glycerogelatin, etc. may be used.

[0151] The above pharmaceutical composition may have any one formulation selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid formulations, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories.

[0152] The pharmaceutical composition of the present invention can be administered to a subject in a pharmaceutically effective amount.

[0153] In the present invention, the term "pharmaceuticalally effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including individual type and severity, age, gender, type of brain disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. It may also be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by considering all of the above factors, and this can be easily determined by a person skilled in the art.

[0154] In the present invention, the term "object" is used interchangeably with "patient" and "individual," and may be a mammal requiring prevention or treatment of a degenerative brain disease, e.g., primates (e.g., humans), companion animals (e.g., dogs, cats, etc.), livestock animals (e.g., cattle, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). In one embodiment of the present invention, the object may be a human.

[0156] Another aspect of the present invention provides a method for preventing or treating a degenerative brain disease, comprising the step of administering one or more of an anti-CD300c antibody and an antigen-binding fragment thereof to a subject having a degenerative brain disease or at risk thereof.

[0157] The above anti-CD300c antibody, its antigen-binding fragment, degenerative brain disease, subject, prevention, and treatment, etc., are as described in other embodiments.

[0158] In the present invention, the term "administration" means a method of providing a substance (e.g., any one or more of an anti-CD300c antibody and its antigen-binding fragments) to a subject for a preventive or therapeutic purpose (e.g., prevention or treatment of a degenerative brain disease).

[0159] The antibody and / or antigen-binding fragment of the present invention may be administered in various ways depending on whether local or systemic treatment is desired and the area to be treated. The method of administering the antibody and / or antigen-binding fragment of the present invention to a subject can be easily determined by those skilled in the art. The route of administration may be oral, parenteral, inhaled, or local. For example, parenteral administration may include, but is not limited to, intravenous administration, subcutaneous administration, intraperitoneal administration, intramuscular administration, rectal administration, vaginal administration, intrathecal administration, intraventricular administration, or intraventricular administration. In one embodiment, the antibody and / or antigen-binding fragment of the present invention may be delivered across the blood-brain barrier (BBB) ​​using various suitable compositions and methods described herein or known in the art.

[0160] The effective or effective no-toxic amount of the antibody and / or antigen-binding fragment of the present invention can be determined by conventional experiment. For example, the therapeutically active amount of the antibody and / or antigen-binding fragment of the present invention may vary depending on factors such as the stage of the disease, the severity of the disease, the age, sex, medical complications, and body weight of the subject, and the ability of the antibody to induce the desired response in the subject. The dosage and administration regimen of the antibody and / or antigen-binding fragment of the present invention may be adjusted to provide an optimal therapeutic response. For example, several divided doses may be administered daily, weekly, every two weeks, every three weeks, every four weeks, etc., or the dose may be proportionally reduced or increased depending on the urgency of the treatment situation.

[0161] The antibody and / or antigen-binding fragment of the present invention may be administered in combination with one or more other agents effective for the prevention or treatment of neurodegenerative diseases. The other agents may include, but are not limited to, any agents capable of improving or resolving neurodegenerative diseases, such as compounds, gene therapies, and proteins (including antibodies). When the antibody and / or antigen-binding fragment of the present invention is administered together with other agents, they may be formulated as a single composition for simultaneous delivery, or they may be formulated individually into two or more compositions (e.g., a kit). Each component may be administered to a subject at a different time than when the other components are administered. In a specific embodiment, each administration may be given non-simultaneously (e.g. individually or sequentially) at multiple intervals over a given period. Additionally, individual components may be administered to a subject by the same or different routes. The pharmaceutical composition of the present invention may be used in combination with one or more previously known treatments for neurodegenerative diseases.

[0163] Another aspect of the present invention provides a food composition for preventing or improving degenerative brain diseases comprising any one or more of an anti-CD300c antibody and an antigen-binding fragment thereof.

[0164] The above anti-CD300c antibody, its antigen-binding fragment, degenerative brain disease, and prevention, etc., are as described in other embodiments.

[0165] The above food composition may be a health functional food.

[0166] In the present invention, the term "improvement" refers to any act of using any one or more of the anti-CD300c antibody and its antigen-binding fragments to reduce the survival, recurrence, and / or resistance to therapeutic agents of individuals suspected of or suffering from neurodegenerative disease, or to improve or benefit symptoms.

[0167] In the present invention, the term "food" includes all foods in the conventional sense, such as meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, and health functional foods, and is not limited thereto as long as it may include one or more of the anti-CD300c antibody and its antigen-binding fragments according to the present invention. When preparing the above food composition, it may be prepared by adding raw materials and ingredients that are conventionally added in the art, and the types thereof are not particularly limited. For example, as with conventional foods, various herbal extracts, food-grade acceptable food additives, or natural carbohydrates may be included as additional ingredients, but are not limited thereto. The mixing amount of the active ingredients may be appropriately determined according to the purpose of use.

[0168] In the present invention, the term "health functional food" is synonymous with "food for special health use (FosHU)," and refers to a food manufactured or processed for the purpose of health supplementation by using specific ingredients as raw materials or by methods such as extraction, concentration, purification, or mixing of specific ingredients contained in food raw materials. It refers to a food designed and processed to fully exert bio-regulatory functions on the body, such as biological defense, regulation of biological rhythms, and prevention and recovery from disease, through the aforementioned ingredients; and the composition for the health functional food can perform functions related to the prevention and recovery from disease. The health functional food of the present invention may be used interchangeably with terms known in the art, such as "functional food."

[0170] Another aspect of the present invention provides a feed composition for preventing or improving degenerative brain diseases comprising any one or more of an anti-CD300c antibody and an antigen-binding fragment thereof.

[0171] The above anti-CD300c antibody, its antigen-binding fragment, prevention of degenerative brain disease, and improvement, etc., are as described in other embodiments.

[0172] The above feed composition may be for animals other than humans, and may be for said individuals, such as mammals including cattle, pigs, sheep, chickens, dogs, humans, etc., birds, etc., but is not limited thereto.

[0173] In addition to the anti-CD300c antibody and its antigen-binding fragment of the present invention, the above feed composition may include known carriers, stabilizers, or additives acceptable for pharmaceutical, food, or feed purposes. For example, binders, emulsifiers, preservatives, etc. added to prevent quality degradation, and amino acid preparations, vitamins, enzymes, flavorings, non-protein nitrogen compounds, silicates, buffers, extractants, oligosaccharides, etc. added to the feed to enhance utility. In addition, feed mixing agents, etc., may be additionally included, but are not limited thereto. The above feed composition may include various nutrients such as vitamins, amino acids, and minerals, antioxidants, and other additives as needed, and may be in a suitable form such as powder, granules, pellets, or suspension. The feed composition of the present invention may be supplied to monogastric animals alone or mixed with feed. The feed of the present invention is not particularly limited and any feed such as powder feed, solid feed, dry feed, wet feed, moist pellet feed, dry pellet feed, EP (Extruder Pellet) feed, raw feed, etc. may be used.

[0175] Another aspect of the present invention provides the use of an anti-CD300c antibody and / or an antigen-binding fragment thereof for the prevention or treatment of degenerative brain diseases.

[0176] In one embodiment, the above-mentioned degenerative brain disease may be prevented or treated by promoting differentiation from monocytes into MDM or M1 macrophages.

[0177] Another aspect of the present invention provides an anti-CD300c antibody and / or an antigen-binding fragment thereof for use in manufacturing a composition (medicine, etc.) for the prevention or treatment of degenerative brain diseases.

[0178] The above anti-CD300c antibody, its antigen-binding fragment, degenerative brain disease, prevention, and treatment, etc., are as described in other embodiments.

[0180] Another aspect of the present invention provides a method for promoting differentiation from monocytes into MDM or M1 macrophages, comprising the steps of administering one or more of an anti-CD300c antibody and its antigen-binding fragments to a subject; or treating isolated cells.

[0181] The above subject may have a degenerative brain disease or be at risk thereof, but is not limited thereto.

[0182] Another aspect of the present invention provides the use of an anti-CD300c antibody and / or its antigen-binding fragment to promote differentiation into MDM or M1 macrophages.

[0183] The above method and / or use for promoting differentiation into MDM or M1 macrophages is in vitro or in vivo It could be.

[0184] The above anti-CD300c antibody, its antigen-binding fragment, the target, etc. are as described in other embodiments. Effects of the invention

[0186] According to the present invention, an antibody that specifically binds to CD300c and its antigen-binding fragment can be effectively used as a preventive or therapeutic agent for degenerative brain diseases.

[0187] Through the Y-maze test and the Morris water maze test using 5xFAD mice mutated to express genes related to neurodegenerative diseases, it was confirmed that the anti-CD300c antibody or its antigen-binding fragment significantly restored cognitive ability and memory in a mouse model of neurodegenerative diseases. Therefore, the anti-CD300c antibody or its antigen-binding fragment has an effective drug effect for the prevention or treatment of neurodegenerative diseases. Brief explanation of the drawing

[0189] Figure 1 shows the results of confirming the binding power of an anti-CD300c monoclonal antibody to a CD300c antigen using a conjugation ELISA. Figure 2 shows the results of a sequence comparison between the heavy chain variable regions of the anti-CD300c monoclonal antibodies CB201, CL10, and SL18. Figure 3 shows the results of confirming the binding power of an anti-CD300c monoclonal antibody to a CD300c antigen using surface plasmon resonance (SPR). Figure 4 is a figure showing the results of confirming the binding power of the anti-CD300c monoclonal antibody to the CD300c antigen of cells. Figure 5 is a figure confirming the results of promoting the differentiation ability of MDM (Monocyte-derived macrophage) by anti-CD300c monoclonal antibody by observing cell morphology. Figure 6 is a figure confirming the results of promoting MDM differentiation ability of the anti-CD300c monoclonal antibody through the analysis of factors involved in cell signaling. Figure 7 is a figure confirming the results of promoting MDM differentiation ability of the anti-CD300c monoclonal antibody CB201 through the measurement of differentiation marker (TNF-α, IL-1β, and IL-8) production. Figure 8 is a figure confirming the results of promoting MDM differentiation ability of anti-CD300c monoclonal antibodies CL10 and SL18 through the measurement of differentiation marker (TNF-α) production. Figure 9 is a figure confirming the results of promoting MDM differentiation ability of the anti-CD300c monoclonal antibody through extracellular protein marker analysis. Figure 10 is a figure confirming the results of promoting MDM differentiation ability by the anti-CD300c monoclonal antibody through marker analysis. (in vivo ). Figure 11 is a figure confirming the results of promoting MDM differentiation ability by the anti-CD300c monoclonal antibody through marker analysis. (in vivo ). Figure 12 is a figure confirming the promotion of MDM differentiation ability and changes in the immune system by the anti-CD300c monoclonal antibody through nanostring immune profiling. Figure 13 shows the animal model construction and experimental method for the Y-maze test. Figure 14 is a figure showing a scene from a video taken during the Morris underwater maze test. Figure 15 is a figure quantifying the results of the Y-maze test and the Morris underwater maze test. Specific details for implementing the invention

[0190] The present invention will be explained in more detail below through examples. However, the following examples are merely preferred embodiments for illustrating the present invention and are therefore not intended to limit the scope of the present invention. Meanwhile, technical matters not described in this specification can be fully understood and easily implemented by a person skilled in the art in the field of the present invention or a similar field.

[0192] Example 1. Preparation of anti-CD300c antibodies (CB201, CL10, and SL18)

[0193] An antibody expression vector was constructed by separating the heavy and light chains using a nucleotide sequence encoding an anti-CD300c monoclonal antibody that specifically binds to the CD300c antigen with high binding affinity.

[0194] To this end, genes for expressing the heavy and light chains were inserted into the pCIW3.3 vector using the CB201 CDR sequences (the amino acid sequence of CB201 and the polynucleotide sequence encoding it) shown in Tables 3 and 4 and the sequence list. A known method was used for vector construction. The constructed CB201 heavy and light chain expression vectors were mixed with PEI (polyethylenimine) in a 1:1 mass ratio and transfected into 293T (human colorectal cancer cell line) cells to induce antibody expression. On day 8, the culture medium was centrifuged to remove the cells, and the culture medium was obtained. The obtained culture medium was filtered and then resuspended in a solution containing a mixture of 0.1 M NaH2PO4 and 0.1 M Na2HPO4 (pH 7.0). The resuspended solution was purified by affinity chromatography using protein A beads (GE healthcare) and finally eluted using elution buffer (Thermofisher).

[0196] antibodies Types of waste paper domain DNA sequences (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Sequence number CB201 VH3VL1 heavy chain variable region entire GAGGTGCAGCTGTTGGAGTCTGGTGGAGGCTTGGTACAGCCTGGAGGTTCTCTTCGCCTCTCCTGTGCAGCCTCCGGATTCACTTTC AGCCGCTACGCAATGAGCTGGGTCAGA CAGGCACCAGGTAAGGGACTGGAGTGGGTCTCT GCAATTAGCGGTAGCGGTGGTAGCACTTACTACGCAGAC AGCGTGAAGGGTCGCTTCACCATCTCACGCGACAACTCCAAGAACACCCTGTACCTGCAGATGAACAGCCTTCGCGCAGAGGACACTGCCGTGTATTAC TGCGCACGTAGCAGCCAGGGTATCTTCGACATCTGGGGA CAAGGTACTCTGGTCACTGTCTCCTCA 325 CDR1 AGCCGCTACGCAATGAGCTGGGTCAGA 73 CDR2 GCAATTAGCGGTAGCGGTGGTAGCACTTACTACGCAGAC 74 CDR3 TGCGCACGTAGCAGCCAGGGTATCTTCGACATCTGGGGA 75 Light chain variable region entire CAGTCTGTGCTGACTCAGCCACCTTCAGCATCTGGTACTCCAGGTCAGCGCGTCACCATCAGC TGCAGTGGTAACAATATCGGTACTAGACGCGTG CATTGGTATCAGCAACTCCCAGACACCGCTCCTAAGCTCCTGATTTACAGT AAGAACAACCGTCCTAGTGGTGTG CCTGATCGCTTTTCTGGGTCCAAGTCTGGCACCTCAGCCTCTCTGGCTATCAGTGGACTTCGCTCCGAGGACGAGGCTGACTATTAC TGCGCAGCATGGGACGACAGCCTGAGCGGTCCTGTGTTC GGCGGTGGGACCAAACTGACCGTCCTA 326 CDR1 TGCAGTGGTAACAATATCGGTACTAGACGCGTG 76 CDR2 AAGAACAACCGTCCTAGTGGTGTG 77 CDR3 TGCGCAGCATGGGACGACAGCCTGAGCGGTCCTGTGTTC 78

[0197] antibodies Types of waste paper domain Amino acid sequence (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Sequence number CB201 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSRYAMSWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSSQGIFDIW GQGTLVTVSS 327 CDR1 FTFSRYAMSWVR 79 CDR2 AISGSGGSTYYAD 80 CDR3 YCARSSQGIFDIW 81 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGTRRVHW YQQLPDTAPKLLIY SKNNRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCAAWDDSLSGPVF GGGTKLTVL 328 CDR1 CSGNNIGTRRVHW 82 CDR2 SKNNRPSGVP 83 CDR3 YCAAWDDSLSGPVF 84

[0198] Anti-CD300c monoclonal antibodies CL10 and SL18 were also prepared in the same manner as CB201 using Tables 5 to 8 below.

[0200] antibodies Types of waste paper domain DNA sequences (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Sequence number CL10 VH3VL1 heavy chain variable region entire GAGGTGCAGCTGTTGGAGTCTGGTGGAGGCTTGGTACAGCCTGGAGGTTCTCTTCGCCTCTCCTGTGCAG CCTCCGGATTCACTT TCAGCAGCTACGGTATGCATTGGGTCAGACA GGCACCAGGTAAGGGACTGGAGTGGGTCT CTGCAATTAGCGGTAGCGGTGGTAGCACTTACTACGCAGACAG CGTGAAGGGTCGCTTCACCATCTCACGCGACAACTCCAAGAACACCCTGTACCTGCAGATGAACAGCCTTCGCGCAGAGGACACTGCCGTGTATT ACTGCGCAAGCGGTTACGGTCTGATGGACGTTGTGGGGACA AGGTACTCTGGTCACTGTCTCCTCA 337 CDR1 TCAGCAGCTACGGTATGCATTGGGTCAGACA 109 CDR2 CTGCAATTAGCGGTAGCGGTGGTAGCACTTACTACGCAGACAG 110 CDR3 ACTGCGCAAGCGGTTACGGTCTGATGGACGTTGTGGGGACA 111 Light chain variable region entire TCTGTGCTGACTCAGCCACCTTCAGCATCTGGTACTCCAGGTCAGCGCGTCACCATCA GCTGCACTCGTAGCAGCGGTATCATCGCAAGCAACTACGTGCA GTGGTATCAGCAACTCCCAGGCACCGCTCCTAAGCTCCTGATTTAC CGCAACAACCAGCGCCCTAGTGGTGTG CCTGATCGCTTTTCTGGGTCCAAGTCTGGCACCTCAGCCTCTCTGGCTATCAGTGGACTTCGCTCCGAGGACGAGGCTGACTATT ACTGCAGCAGCTACGCAGGTAACAACAACCTGGTGTTCGG CGGTGGGACCAAACTGACCGTCCTA 338 CDR1 GCTGCACTCGTAGCAGCGGTATCATCGCAAGCAACTACGTGCA 112 CDR2 CGCAACAACCAGCGCCCTAGTGGTGTG 113 CDR3 ACTGCAGCAGCTACGCAGGTAACAACAACCTGGTGTTCGG 114

[0201] antibodies Types of waste paper domain Amino acid sequence (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Amino acid sequence number CL10 VH3VL1 heavy chain variable region entire EVQLLESGGGLVQPGGSLRLSCAASG FTFSSYGMHWVR QAPGKGLEWVS AISGSGGSTYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCASGYGLMDVW GQGTLVTVSS 339 CDR1 FTFSSYGMHWVR 115 CDR2 AISGSGGSTYYAD 116 CDR3 YCASGYGLMDVW 117 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CTRSSGIIASNYVQW YQQLPGTAPKLLIY RNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADY YCSSYAGNNNLVF GGGTKLTVL 340 CDR1 CTRSSGIIASNYVQW 118 CDR2 RNNQRPSGVP 119 CDR3 YCSSYAGNNNLVF 120

[0202] antibodies Types of waste paper domain DNA sequences (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Sequence number SL18 Lambda heavy chain variable region entire CGAGTGCAGCTGCTGGAAAGTGGAGGTGGACTGGTGCAGCCTGGCGGCAGCCTGCGCCTGAGCTGTGCCG CCAGCGGAT TCACCTTCAGCGATTATCATATGCATTGGGTTCG CCAAGCACCTGGCAAAGGCCTGGAATGGGTGAGCACCA TCAGCAGCAGCGGCGGCTATACCTATTATGCCGA AAGCGTGAAAAGCCGCTTTACCATCAGCCGCGATAACAGCAAAAACACCCTGTATCTGCAGATGAACAGCCTGCGCGCCGAGGACACCGCAGTCTACTACTGTG CCCGATCGATACGCCTGCCTCTGGATTATTGGGG ACAAGGTACTCTGGTGACCGTGAGCAG CA 369 CDR1 TCACCTTCAGCGATTATCATATGCATTGGGTTCG 205 CDR2 TCAGCAGCAGCGGCGGCTATACCTATTATGCCGA 206 CDR3 CCCGATCGATACGCCTGCCTCTGGATTATTGGGG 207 Light chain variable region entire CAGAGCGTGCTGACCCAGCCTCCTAGCGCCTCCGGTACACCAGGACAGCGCGTGACTATTAGCTGTA GCGGCAACAACATCGGCAGCAAAGGCGTGCATTGGTA TCAGCAACTGCCTGGAACTGCACCTAAGCTGCTGATCT ATGAAGATAGCAAACGCCCTAGCGGCGTGCGTGA TCGCTTTAGCGGTAGCAAATCAGGCACCAGCGCCAGCCTGGCCATCAGCGGCCTTCGCTCCGAAGATGAAGCCGATTATTATTGTCAGA GCTATGATAGCACCAAAGGCGTGGTGTTTGGTGG CGGTACCAAGC GACCGTGCTG 370 CDR1 GCGGCAACAACATCGGCAGCAAAGGCGTGCATTGGTA 208 CDR2 ATGAAGATAGCAAACGCCCTAGCGGCGTGCGTGA 209 CDR3 GCTATGATAGCACCAAAGGCGTGGTGTTTGGTGG 210

[0203] antibodies Types of waste paper domain Amino acid sequence (CDR1, CDR2, and CDR3 are shown in bold and underlined in order) Amino acid sequence number SL18 Lambda heavy chain variable region entire RVQLLESGGGLVQPGGSLRLSCAASG FTFSDYHMHWVR QAPGKGLEWVS TISSSGGYTYYAE SVKSRFTISRDNSKNTLYLQMNSLRAEDTAVY YCARSIRLPLDYW GQGTLVTVSS 371 CDR1 FTFSDYHMHWVR 211 CDR2 TISSSGGYTYYAE 212 CDR3 YCARSIRLPLDYW 213 Light chain variable region entire QSVLTQPPSASGTPGQRVTIS CSGNNIGSKGVHW YQQLPGTAPKLLIY EDSKRPSGVR DRFSGSKSGTSASLAISGLRSEDEADY YCQSYDSTKGVVF GGGTKLTVL 372 CDR1 CSGNNIGSKGVHW 214 CDR2 EDSKRPSGVR 215 CDR3 YCQSYDSTKGVVF 216

[0205] Example 2. Confirmation of binding affinity of anti-CD300c monoclonal antibody to CD300c antigen (I): Conjugation ELISA

[0206] CD300c antigen (250 µg / mL) was diluted to a concentration of 800 ng / mL in coating buffer (0.1 M sodium carbonate, pH 9.0), and 100 µL of this solution was added to each well of a 96-well microplate and incubated overnight at 4°C. The next day, the microplate was washed three times with 200 µL of PBST. Afterward, 200 µL of blocking buffer (5% skim milk) was added to each well and blocked at room temperature for 1 hour.

[0207] The anti-CD300c monoclonal antibody CB201 was diluted to 200 µg / mL in PBS, and the concentration was verified using a Nanodrop (product name: NanoDrop One / Onec). Subsequently, CB201 was quadrupled from 10 µg / mL in PBS, and 100 µL was added to each well. The mixture was then incubated at room temperature for 1 hour and washed three times with 200 µL of PBST. 100 µL of the secondary antibody (conjugated anti-Fc IgG), diluted 1:10,000 in blocking buffer, was added to each well and incubated at room temperature for 1 hour. Afterward, 200 µL of PBST was added and washed three times. Next, TMB and hydrogen peroxide were mixed in a 1:1 ratio, and 100 µL was added to each well. The mixture was then incubated at room temperature for 7 to 9 minutes. After that, 50 µl of 1 N sulfuric acid was added to stop the color development, and the binding affinity results were obtained by measuring the value appearing at 450 nm using a microplate reader (product name: Varioskan LUX).

[0208] In addition, the anti-CD300c monoclonal antibodies CL10 and SL18 also obtained binding affinity results in the same manner as CB201.

[0210] As a result, as shown in Figure 1, it was confirmed that the anti-CD300c monoclonal antibodies CB201, CL10, and SL18 all bind to CD300c in a concentration-dependent manner, which indicates that the anti-CD300c monoclonal antibodies CB201, CL10, and SL18 all have excellent binding affinity and specificity for the antigen CD300c.

[0212] From the above results, it was confirmed that the anti-CD300c monoclonal antibodies not only have excellent binding affinity and specificity for CD300c, but antibodies CB201, CL10, and SL18 are all anti-CD300c monoclonal antibodies with similar properties, which is due to the similar variable region sequences of the three antibodies as shown in Figure 2.

[0214] Example 3. Confirmation of binding affinity of anti-CD300c monoclonal antibody to CD300c antigen (II): Surface plasmon resonance (SPR)

[0215] Surface plasmon resonance experiments were conducted to further confirm the binding affinity between the antigen CD300c and the anti-CD300c monoclonal antibody CB201.

[0216] To immobilize CD300c on a CM5 chip, 5 µg / ml of CD300c was diluted in 10 mM acetate buffer (pH 5.5). Then, the flow rate was set to 10 ml / min for all samples, and the target RU (Resonance Units) for each sample was set to 300 RU. The CM5 chip was activated with a mixture of 0.2 M EDC and 0.05 M NHS and blocked with 1 M ethanolamine to immobilize the CM5 chip such that the final RU of CD300c was 399.2 RU. Subsequently, CB201 was diluted in PBST to concentrations of 0, 0.195, 0.39, 0.78, 1.56, 3.125, and 6.25 µg / ml, respectively, and a kinetics / affinity test was performed with a binding time of 240 seconds, a dissociation time of 900 seconds, and a flow rate of 30 µl / min. Afterward, the surface was regenerated by flowing 50 mM NaOH over the CD300c-immobilized CM5 chip at a rate of 30 µl / min for 30 seconds.

[0217] As a result, as shown in Figure 3, the KD value was analyzed to be 5.199E-10 M, and the binding affinity of the anti-CD300c monoclonal antibody was confirmed to be 0.52 nM, which is a subnanomol level. This means that the binding power of the anti-CD300c monoclonal antibody to the antigen is high.

[0219] Example 4. Confirmation of Cell Antigen Recognition by Anti-CD300c Monoclonal Antibody

[0220] FACS binding was performed to confirm that the anti-CD300c monoclonal antibody (CB201) recognizes cell antigens.

[0221] To this end, CD300c was overexpressed in 293T cells (ATCC), and then these cells were [strained] 2 x 10⁶ per microcentrifuge tube 5The cells were dispensed. Then, anti-CD300c monoclonal antibodies serially diluted three-fold starting from 10 µg / ml were reacted with the cells in a CO2 incubator for 30 minutes, and the tubes were washed twice with FACS buffer. Subsequently, FITC-conjugated anti-human IgG(H+L) diluted 1:100 in FACS buffer was reacted with the anti-CD300c monoclonal antibodies bound to the cells in a CO2 incubator for 30 minutes, and the tubes were washed twice with FACS buffer. Next, the FITC signal was measured using a Beckman Coulter CytoFLEX instrument, and the MFI value was calculated using the CytExpert program. Using the obtained MFI value, a sigmoidal curve was plotted using the sigmaplot program to calculate the EC50 (The effective concentration of drug that causes 50% of the maximum response). As a result, an EC50 of 2.7 nM was obtained for 293T cells.

[0222] As shown in Figure 4, in the S-shaped curve resulting from FACS binding, the anti-CD300c monoclonal antibody bound to CD300c overexpressed on the surface of 293T cells with strong binding affinity. Therefore, it was confirmed that the anti-CD300c monoclonal antibody binds antigen-specifically to CD300c even at the cellular level.

[0224] Example 5. Confirmation of promotion of differentiation ability into MDM (Monocyte-derived macrophage) (I): Observation of cell morphology

[0225] To confirm the morphological pattern of differentiation from monocytes to MDM (Monocyte-derived macrophage) when monocytes were treated with an anti-CD300c monoclonal antibody, THP-1 cells (human monocyte cell line) treated with 10 μg / ml of the anti-CD300c monoclonal antibody CB201 were cultured for 48 hours, after which the cell morphology was observed under a microscope.

[0226] As a result, as shown in Figure 5, it was confirmed that in the experimental group (CB201) treated with the anti-CD300c monoclonal antibody, the morphology of THP-1 cells changed from suspension cells to round adherent cells in the form of MDM.

[0227] Through the above results, it was confirmed that treatment with the anti-CD300c monoclonal antibody promotes differentiation from monocyte cells into MDM.

[0229] Example 6. Confirmation of Promotion of Differentiation Ability into MDM (II): Cell Signaling

[0230] To further confirm that the differentiation ability from monocytes to MDMs increases when THP-1 cells are treated with CB201, an anti-CD300c monoclonal antibody, the signaling of MAPK (mitogen-activated protein kinase) and NF-kB, which are representative signals of MDM differentiation, was examined.

[0231] To this end, 8.8 x 10⁻⁶ in a 6-well plate 5 THP-1 cells were dispensed into wells and treated with 10 μg / mL of anti-CD300c monoclonal antibody. As a control, an equal amount of phosphate buffer (PBS) was injected. After incubating them for 48 hours, phosphorylated SAPK / JNK, phosphorylated ERK (p44 / 42), and phosphorylated p38 for MAPK signaling, and phosphorylated NF-kB for NF-kB signaling were confirmed by Western blotting.

[0232] From this, the results confirming the signal transduction of MAPK and NF-Kb, respectively, are shown in Figure 6. It was confirmed that the amount of phosphorylated MAPK and NF-kB increased when anti-CD300c was treated compared to the control group.

[0233] Through the above results, it was confirmed that cell signaling for differentiation into MDM increased when treated with the anti-CD300c monoclonal antibody.

[0235] Example 7. Comparison of the differentiation ability of anti-CD300c monoclonal antibody into MDM: Measurement of production of three differentiation markers (TNF-α, IL-1β, and IL-8).

[0236] To confirm the MDM differentiation ability of the anti-CD300c monoclonal antibody and other antibodies, 8.8 x 10⁶ in a 6-well plate 5 THP-1 cells were dispensed into wells, and treated with 10 μg / mL of the anti-CD300c monoclonal antibody CB201. The production of MDM differentiation markers TNF-α, IL-1β, and IL-8 was confirmed using an ELISA kit. As another antibody, a human IgG isotype control was treated at a concentration of 10 μg / mL, and the production of TNF-α, IL-1β, and IL-8 was confirmed using an ELISA kit.

[0237] In addition, anti-CD300c monoclonal antibodies CL10 and SL18 were also treated to THP-1 cells in the same manner as CB201, and the amount of TNF-α produced was confirmed using an ELISA kit, and the results are shown in Figure 7 (CB201) and Figure 8 (CL10 and SL18).

[0238] As shown in Figure 7, it was confirmed that the anti-CD300c monoclonal antibody CB201 significantly increased the production of TNF-α, IL-1β, and IL-8 compared to the control group treated with the isotype control alone. Similarly, as shown in Figure 8, it was confirmed that the anti-CD300c monoclonal antibodies CL10 and SL18 significantly increased the production of TNF-α compared to the control group treated with the isotype control alone. This suggests that the anti-CD300c monoclonal antibodies CL10 and SL18 have similar characteristics to CB201.

[0239] Through the above results, it was confirmed that the anti-CD300c monoclonal antibody significantly increases the differentiation ability of monocytes into MDMs.

[0241] Example 8. Confirmation of Promotion of Differentiation Ability into MDM by Anti-CD300c Monoclonal Antibody (III): Identification of Extracellular Protein Expression Markers

[0242] To confirm the effect of CB201, an anti-CD300c monoclonal antibody, on differentiating monocytes into MDMs, changes in the expression of proteins that are MDM differentiation markers were examined via FACS in THP-1 cells treated with CB201 in the same manner as in Example 6. As a result of examining the changes in expression using respective antibodies that recognize the monocyte marker CD11b, the MDM marker CD80, and the M2 macrophage marker CD206, it was confirmed that the MDM marker CB80 increased in cells treated with CB201, as shown in Figure 9.

[0243] From the above results, it was confirmed that treating monocytes with CB201, an anti-CD300c monoclonal antibody, increased the expression of the MDM marker among extracellular proteins. Through this, it was confirmed that CB201 differentiates monocytes into MDM.

[0245] Example 9. Preparation of a mouse model of a degenerative brain disease.

[0246] Eight male and eight female 4-8 week old 5xFAD mice (source: Jackson lab) and three male and three female 6-week old B6SJLF1 / J mice (source: Jackson lab) were purchased and their health status was confirmed by observing general symptoms after an acclimatization period of about 4-5 months, and healthy animals were used in the experiment. 5xFAD mice are mice mutated to express human APP (Amyloid Precursor Protein) and PSEN1 genes that cause a total of five AD-related mutations (Swedish (K670N / M671L), Florida (I716V), London (V717I) mutations of APP, and M146L, L286V mutations of PSEN1), and were selected as a suitable model for pathological studies related to neuronal damage diseases, including Alzheimer's disease symptoms as well as dementia (Tatsuhiro Ayabe et al, The Neuroscience of Dementia, 2020, Vol. 2, 833-847). The experiment was conducted in Animal Rearing Area No. 3 of Notus Co., Ltd., with environmental conditions set to a temperature of 23±3℃, relative humidity of 55±15%, ventilation rate of 10–20 times / hr, lighting time of 12 hours (lights turned on at 8:00 AM to off at 8:00 PM), and illuminance of 150–300 Lux. The experimental animals were classified into three groups: a normal mouse group (G1), a degenerative brain disease mouse group (hereinafter, 5xFAD control group; G2), and a CB201-administered degenerative brain disease mouse group (hereinafter, CB201 administration group; G3). The CB201 administration group was administered CB201 at a dose of 40 mg / kg, while the control groups (normal mouse group and 5xFAD control group) were administered PBS at an equal volume. The test substance was administered intraperitoneally a total of two times: on the test substance start date (Day 0) and on the 25th day after the start of administration (Day 25). Refer to Table 9 below for the mouse groups used in the study.

[0248] army gender Number of animals (parts) Animal number system Administered substance Dosage (mg / kg) Administered volume (mL / kg) Normal mouse (G1) M / F 3 / 3 1-3 / 12-14 B6SJLF1 / J Vehicle - 10 Degenerative brain disease mice (5xFAD control group, G2) M / F 4 / 4 4-7 / 15-18 5xFAD Vehicle - 10 CB201 administration group (G3) M / F 4 / 4 8-11 / 19-22 5xFAD CB201 40 10

[0250] Example 10. In vivo ( in vivo Confirmation of the effect of anti-CD300c monoclonal antibody on increasing MDM in ).

[0251] To determine whether the anti-CD300c monoclonal antibody increases MDM in a mouse model, brain tissue was extracted from mice administered CB201 by the method of Example 9 (CB201 administration group; G3), and the ratio of MDM in the brain tissue was determined using antibodies against iNOS, an MDM marker, and CD206, an M2 macrophage marker.

[0252] In addition, an experimental group of mice administered an anti-PD-1 antibody was also prepared by administering 40 mg / kg of PD-1 antibody instead of 40 mg / kg of CB201 to 5xFAD mice in the same manner as the preparation of the CB201 administration group in Example 9.

[0253] As a result, as shown in Figure 10, in the brain tissue of the experimental group administered an anti-PD-1 antibody, which is known to have an effect of improving symptoms of neurodegenerative disease as an immunotherapeutic agent, MDM was slightly increased (iNOS was slightly increased) compared to the control group (neurodegenerative brain disease mice in Table 9; G2), but in the brain tissue of the experimental group administered an anti-CD300c monoclonal antibody (CB201) (CB201 administration group in Table 9; G3), MDM was significantly increased (iNOS was significantly increased) compared to the control group and M2 macrophages were hardly observed.

[0254] From the above results, it was confirmed that the anti-CD300c monoclonal antibody CB201 has a significant effect in differentiating monocytes into MDMs compared to other immunotherapies.

[0256] Example 11. Confirmation of increased expression of MDM differentiation genes by anti-CD300c monoclonal antibody

[0257] To confirm the effect of the anti-CD300c monoclonal antibody CB201 on brain MDM under in vivo conditions, brain tissue was extracted from mice in the CB201 administration group (G3 in Table 9) prepared as in Example 9, and the cells were stained with a stain for confirming cell viability (DAPI), a total monocyte marker F4 / 80, and an antibody against the MDM marker CD11b (Source: Invitrogen). Subsequently, the data was read using a CytoFLEX flow cytometer, and the resulting data was analyzed using FlowJo software.

[0258] As a result of quantifying each marker relative to surviving cells, as shown in Fig. 11, it was confirmed that in mouse brain tissue treated with the anti-CD300c monoclonal antibody alone compared to the control group, the expression level of the monocyte marker F4 / 80 was similar, while the expression level of the MDM differentiation marker CD11b increased (corresponding to M1 MDM in CD11b+ in Fig. 11).

[0259] As described below, it has been confirmed that the anti-CD300c monoclonal antibody has a therapeutic effect on neurodegenerative diseases. The aforementioned results suggest that the therapeutic effect on neurodegenerative diseases is achieved by increasing MDM differentiation in brain tissue through the administration of the anti-CD300c monoclonal antibody.

[0261] Example 12. Nanostring Immunoprofiling

[0262] To confirm changes in immune cells and the brain tissue environment within a mouse model of neurodegenerative disease when an anti-CD300c monoclonal antibody (CB201) was administered, brain tissue was extracted from mice in the CB201 administration group (G3 in Table 9) prepared as in Example 9. RNA was extracted and purified from the tissue, and changes in monocyte markers were confirmed through nanostring immune profiling, a known method. The results observed in comparison with the control group are shown in Figure 12.

[0263] As shown in Figure 12, upon administration of the anti-CD300c monoclonal antibody, it was confirmed that the expression of MDM differentiation markers CD163, Siglec1, Lyve1, and C4b increased, and the expression of IL-10, which exacerbates the progression of neurodegenerative diseases in the brain, decreased. Through this, it was confirmed that the brain's immune system changes in a direction that leads to the differentiation of MDM in brain tissue and the treatment of neurodegenerative diseases.

[0265] Example 13. Y-maze test

[0266] We purchased 6-week-old 5xFAD mice and, after an acclimatization period of about 5 months, developed a neurodegenerative disease model that exhibits symptoms related to neuronal damage, including neurodegenerative disease symptoms.

[0267] Subsequently, 40 mpk of the anti-CD300c monoclonal antibody (CB201) was administered via IP (intraperitoneal) to a neurodegenerative disease model, followed by another administration on the 28th day, and a Y-maze test was performed on the 31st day, while the control group was injected with an equal amount of PBS. The timing of the experiment is shown in Figure 13.

[0268] The experimental apparatus for the Y-maze test consisted of a Y-shaped maze constructed from black acrylic plates (10 cm x 41 cm x 25 cm). The mazes were designated as areas A, B, and C, and experimental animals were placed in random areas and allowed to move freely for 7 minutes. The number of times and the order in which each animal entered each maze were measured to evaluate spontaneous alteration (%). If an animal entered three different areas sequentially, it was counted as 1 point (in the order of alteration, ABC, BCA, CAB, etc.); if it did not enter sequentially, no score was awarded. Spontaneous alteration (% spontaneous alteration) was calculated using the following formula.

[0270]

[0272] As a result, as shown in Figure 15, experimental results showed that while the ability to change was reduced in the mouse model of neurodegenerative disease compared to normal mice (G1; wild type) (G2; 5XFAD+Mock), when CB201 was administered to the mouse model of neurodegenerative disease (G3; 5XFAD+CB201), the ability to change was restored to 87% of the level of normal mice (G1; wild type). The above results suggest that the anti-CD300c monoclonal antibody can be a drug that can alleviate symptoms such as cognitive decline and reduced ability to change that may be caused by neurodegenerative disease.

[0274] Example 14. Morris Underwater Maze Test

[0275] After administering CB201 to mice of a degenerative brain disease model in the same manner as in Example 13, the Morris water maze test was performed on days 35–40. The experimental equipment consisted of a circular tank (stainless steel, diameter 150 cm, height 45 cm), an escape platform (diameter 10 cm, height 30 cm), and four markers attached to the wall to remember the location of the escape platform. The tank was filled with water at a temperature of approximately 22 ± 2 ℃, and the escape platform was positioned either 1 cm above the water surface or 0.5–1.5 cm below the water surface, depending on the experimental purpose. When positioned below the water surface, the water was opaque using white water-based paint so that the escape platform was not visible to the naked eye. The underwater maze was divided into quadrants: Northeast (NE), Northwest (NW), Southeast (SE), and Southwest (SW). An escape platform was placed in the center of the Northeast quadrant, and one of the remaining quadrants was used as the starting position. The experiment was conducted for a total of 6 days, and the starting quadrants and escape platform locations for the animals are indicated in Table 10 below.

[0277] 1 day 2 days 3 days 4 days 5 days 6th Escape location Starting direction Escape location: Southwest, Starting location: As follows No escape squad Test 1 southwest other west north north dong north Test 2 northwest north other west dong other Test 3 northeast other north dong west west Test 4 center dong dong west other dong Test 5 southeast west other other north north Note that on Day 1, both the escape platform location and the starting direction change, whereas on Days 2 through 5, the escape platform location remains constant while the starting direction changes. On Day 6, there is no escape platform, and the test is performed once. Since the starting direction on Day 6 is the farthest from the previous escape platform location (SW), a certain distance must be moved before entering the previously learned escape platform quadrant.

[0278] On Day 1, cued learning was performed by providing a cue by making the escape bar visible. Each individual was attempted 5 times at 15-second intervals, with a time limit of 60 seconds. If the animal found the escape bar within 60 seconds, it was allowed to stay on the escape bar for 5 seconds and then immediately moved to the rearing cage; if it did not find the escape bar within 60 seconds, it was placed on the escape bar for 20 seconds and then immediately moved to the rearing cage.

[0279] From day 2 to day 5, the animals were instructed to memorize a hidden platform by placing the escape platform below the water surface. Each individual attempted this five times at 15-second intervals. A time limit of 60 seconds was set; if the escape platform was found within 60 seconds, the animal was allowed to remain on the platform for 5 seconds and then immediately moved to its rearing cage. If the escape platform was not found within 60 seconds, the animal was placed on the platform for 20 seconds and then immediately moved to its rearing cage.

[0280] On Day 6, a Prospective Randomized Open Blinded End-Point (Probe) Trial was conducted. Without placing an escape platform, the animals were placed in the quadrant furthest from where the escape platform had been located during the experiments on Days 2 through 5, and a single trial was performed with a time limit of 60 seconds. The escape latency—the time it took for the animals to find the escape platform—and the path length to find it were measured and recorded during the cue learning and hidden platform trials. Swimming speed was calculated by dividing the path length by the escape latency. During the probe trial on Day 6, the time the animals spent in the quadrant where the escape platform had previously been located was recorded.

[0281] The aforementioned Y-maze test was filmed as shown in Fig. 14. As shown in Figs. 14 and 15, the experimental results showed that the Time spent in platform quadrants (TSPQ) decreased in the mouse model of neurodegenerative disease compared to normal mice (G1; wild type) (G2; 5XFAD+Mock), whereas in the group administered CB201 to the mouse model of neurodegenerative disease (G3; 5XFAD+CB201), the TSPQ recovered to 107% compared to normal mice (G1; wild type). These results confirm that the administration of CB201 to disease model mice restores cognitive ability and memory in the mouse model of neurodegenerative disease.

[0283] Example 15. Statistical Analysis

[0284] Assuming normality of the data for the test results, a parametric one-way ANOVA was performed. If the results were significant, a post-hoc test was conducted using Dunnett's multiple comparison test to analyze significant differences between the test groups. Statistical analysis was performed using Prism 7.04 (GraphPad Software Inc., San Diego, CA, USA), and a p-value of less than 0.05 was deemed statistically significant.

[0286] From the foregoing description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without altering its technical concept or essential features. In this regard, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as including all modifications or variations derived from the meaning and scope of the claims set forth below and their equivalents, rather than from the detailed description above.

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of a degenerative brain disease comprising, as an active ingredient, one or more of an anti-CD300c antibody and an antigen-binding fragment thereof, wherein one or more of the anti-CD300c antibody and the antigen-binding fragment thereof comprises: (i) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 79, CDR2 composed of SEQ ID NO. 80, and CDR3 composed of SEQ ID NO. 81; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 82, CDR2 composed of SEQ ID NO. 83, and CDR3 composed of SEQ ID NO. 84; (ii) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 115, CDR2 composed of SEQ ID NO. 116, and CDR3 composed of SEQ ID NO. 117; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 118, CDR2 composed of SEQ ID NO. 119, and CDR3 composed of SEQ ID NO.

120. A pharmaceutical composition selected from (iii) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 211, a CDR2 composed of SEQ ID NO. 212, and a CDR3 composed of SEQ ID NO. 213; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 214, a CDR2 composed of SEQ ID NO. 215, and a CDR3 composed of SEQ ID NO.

216. Claim 2 A pharmaceutical composition according to claim 1, wherein the degenerative brain disease is a disease accompanied by cognitive impairment. Claim 3 A pharmaceutical composition according to paragraph 2, wherein the cognitive dysfunction comprises one or more selected from the group including cognitive decline, memory impairment, language impairment, visuospatial decline, orientation impairment, and executive function impairment. Claim 4 A pharmaceutical composition according to claim 1, wherein the degenerative brain disease comprises one or more selected from the group including dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, cerebral amyloid angiopathy, amyloid stroke, Dutch amyloidosis, tauopathy, mild cognitive impairment, corticobasal degeneration, posterior cortical atrophy, primary progressive aphasia, progressive supranuclear palsy, corticobasal deteneration (CBD), amyotrophic lateral sclerosis, Creutzfeldt-Jakob disease, amnesia, learning disability, and memory impairment. Claim 5 A pharmaceutical composition according to claim 4, wherein the dementia comprises one or more selected from the group comprising Alzheimer's disease dementia, Huntington's disease dementia, Parkinson's disease dementia, cerebrovascular dementia, neuroinflammatory dementia, cerebral infarction dementia, senile dementia, Lewy body dementia (Dimentia with Lewy bodies, DLB), Multi-Infarct Dementia (MID), Frontotemporal lobar degeneration (FTLD), Creutzfeldt-Jakob disease dementia, Pick's disease dementia, corticobasal degeneration dementia, dementia due to normal pressure hydrocephalus, and dementia due to head trauma. Claim 6 A pharmaceutical composition according to claim 1, wherein the degenerative brain disease includes Alzheimer's disease. Claim 7 delete Claim 8 delete Claim 9 delete Claim 10 A pharmaceutical composition according to claim 1, wherein at least one of the antibody and the antigen-binding fragments thereof comprises: a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence represented by SEQ ID NO. 79, a CDR2 consisting of an amino acid sequence represented by SEQ ID NO. 80, and a CDR3 consisting of an amino acid sequence represented by SEQ ID NO. 81; and a light chain variable region comprising a CDR1 consisting of an amino acid sequence represented by SEQ ID NO. 82, a CDR2 consisting of an amino acid sequence represented by SEQ ID NO. 83, and a CDR3 consisting of an amino acid sequence represented by SEQ ID NO.

84. Claim 11 delete Claim 12 A pharmaceutical composition according to claim 1, wherein one or more of the antibody and its antigen-binding fragments restore or improve the cognitive function of a subject having a degenerative brain disease or at risk thereof. Claim 13 A method for the prevention or treatment of a neurodegenerative disease, comprising the step of administering one or more of an anti-CD300c antibody and its antigen-binding fragments to a subject other than a human having or at risk of having a neurodegenerative disease, wherein one or more of the anti-CD300c antibody and its antigen-binding fragments comprise: (i) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 79, a CDR2 composed of SEQ ID NO. 80, and a CDR3 composed of SEQ ID NO. 81; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 82, a CDR2 composed of SEQ ID NO. 83, and a CDR3 composed of SEQ ID NO. 84; (ii) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 115, a CDR2 composed of SEQ ID NO. 116, and a CDR3 composed of SEQ ID NO. 117; A method selected from: (iii) a light chain variable region comprising a CDR1 composed of SEQ ID NO. 118, a CDR2 composed of SEQ ID NO. 119, and a CDR3 composed of SEQ ID NO. 120; and (iii) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 211, a CDR2 composed of SEQ ID NO. 212, and a CDR3 composed of SEQ ID NO. 213; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 214, a CDR2 composed of SEQ ID NO. 215, and a CDR3 composed of SEQ ID NO.

216. Claim 14 A food composition for the prevention or improvement of degenerative brain diseases comprising, as an active ingredient, one or more of an anti-CD300c antibody and an antigen-binding fragment thereof, wherein one or more of the anti-CD300c antibody and the antigen-binding fragment thereof comprises: (i) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 79, CDR2 composed of SEQ ID NO. 80, and CDR3 composed of SEQ ID NO. 81; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 82, CDR2 composed of SEQ ID NO. 83, and CDR3 composed of SEQ ID NO. 84; (ii) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 115, CDR2 composed of SEQ ID NO. 116, and CDR3 composed of SEQ ID NO. 117; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 118, CDR2 composed of SEQ ID NO. 119, and CDR3 composed of SEQ ID NO.

120. A food composition selected from (iii) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 211, a CDR2 composed of SEQ ID NO. 212, and a CDR3 composed of SEQ ID NO. 213; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 214, a CDR2 composed of SEQ ID NO. 215, and a CDR3 composed of SEQ ID NO.

216. Claim 15 A feed composition for the prevention or improvement of degenerative brain diseases comprising, as an active ingredient, one or more of an anti-CD300c antibody and an antigen-binding fragment thereof, wherein one or more of the anti-CD300c antibody and the antigen-binding fragment thereof comprises: (i) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 79, CDR2 composed of SEQ ID NO. 80, and CDR3 composed of SEQ ID NO. 81; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 82, CDR2 composed of SEQ ID NO. 83, and CDR3 composed of SEQ ID NO. 84; (ii) a heavy chain variable region comprising CDR1 composed of SEQ ID NO. 115, CDR2 composed of SEQ ID NO. 116, and CDR3 composed of SEQ ID NO. 117; and a light chain variable region comprising CDR1 composed of SEQ ID NO. 118, CDR2 composed of SEQ ID NO. 119, and CDR3 composed of SEQ ID NO.

120. A feed composition selected from (iii) a heavy chain variable region comprising a CDR1 composed of SEQ ID NO. 211, a CDR2 composed of SEQ ID NO. 212, and a CDR3 composed of SEQ ID NO. 213; and a light chain variable region comprising a CDR1 composed of SEQ ID NO. 214, a CDR2 composed of SEQ ID NO. 215, and a CDR3 composed of SEQ ID NO. 216.

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