Anti-βKlotho antibody and its use

Abstract

The present invention relates to a novel anti-βKlotho antibody and its use.The anti-βKlotho antibody can be used for the treatment of diseases mediated by βKlotho or mediated by βKlotho-FGFR1c-FGF21, particularly metabolic disorders, such as type 1 diabetes, type 2 diabetes, dyslipidemia, nonalcoholic fatty liver disease (NASH), nonalcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

Description

[Technical Field]

[0001] (Cross-reference of related applications) This application claims priority to Chinese Patent Application No. 202111683753.3, filed on 30 December 2021, the entire contents of which are incorporated herein by reference for all purposes.

[0002] The present invention generally relates to the field of biopharmaceuticals, and more specifically to novel anti-βKlotho antibodies or their antigen-binding fragments, pharmaceutical compositions comprising the antibody or its antigen-binding fragment, and the medical use of the antibody or its antigen-binding fragment. [Background technology]

[0003] Fibroblast factor 21 (FGF21) is a member of the FGF19 subfamily of the fibroblast factor (FGF) family and plays a crucial role in regulating glucose, lipid, and energy metabolism. In addition to FGF21, the FGF19 subfamily includes two other members: FGF19 and FGF23. Unlike classical fibroblast factors that need to bind to heparin to function, the three proteins of the FGF19 subfamily do not bind to heparin and are secreted into the bloodstream to function physiologically in other tissues or organs of the body. Therefore, these three proteins, including FGF21, are also called endocrine fibroblast factors. FGF21 is mainly expressed in the liver, and also in small amounts in tissues such as the pancreas and brain. It acts on tissues and organs such as adipose tissue and the central nervous system, performing effects such as improving insulin sensitivity, increasing glucose removal from the blood, suppressing hepatic glucose breakdown, improving energy expenditure, increasing hepatic ketone production, and lowering triglyceride levels in the blood. In addition to FGF21, FGF19 is mainly expressed in the small intestine and acts primarily on hepatocytes, involved in regulating bile acid synthesis and secretion in hepatocytes, while FGF23 is a phosphateuria hormone that plays an important role in regulating phosphate balance in humans.

[0004] Three FGF19 subfamily proteins, including FGF21, exert their physiological functions by activating the FGF receptor (FGFR). They cannot activate the FGF receptor alone; they require the assistance of another co-receptor, the Klotho protein, to exert their physiological functions. Klotho proteins are classified into two types, αKlotho and βKlotho, which are necessary for FGF19, FGF21, and FGF23 to bind to their congeneral FGF receptors with high affinity. Here, αKlotho mediates the activation of the FGF receptor by FGF23, while βKlotho mediates the activation of the receptor by FGF19 and FGF21. βKlotho is a single-pass type 1 membrane protein that shares homology with members of family 1 glycosidases but possesses an extracellular domain consisting of two internal repeat sequences that lack glucosidase catalytic activity. βKlotho expression has been detected in tissues such as the liver, pancreas, adipose tissue, and the central nervous system. According to research, β-klotho deficiency (KLB) - / - It was shown that the mRNA levels of CYP7A1 and CY8B1 in mice were elevated, and bile acid synthesis and secretion were increased.

[0005] FGF21 binds to the FGF receptor and forms a ternary complex with β-Klotho. This structure promotes the dimerization of the FGF receptor, forming a hexa-complex that activates the FGF receptor. The activated FGF receptor then activates downstream signaling pathways such as MAPK (Erk1 / 2) and AKT1, thereby exerting its biological function. FGF21 can activate FGF receptors on adipocytes, thereby promoting GLUT-1 expression by adipocytes and increasing glucose uptake by adipocytes. Simultaneously, it can upregulate UCP-1 expression, promoting brown-like transformation of adipocytes, and further induce the expression and secretion of adiponectin or other adipocyte factors that have insulin-sensitizing effects. Adipocyte factors such as adiponectin can further act on other tissues or organs (e.g., the liver) and further improve systemic metabolism. Recent studies have shown that FGF21 also acts on the central nervous system, indirectly regulating glucose, lipid, and energy metabolism, and also influencing the desire for alcohol and sugar intake in humans and mice. Transgenic mice overexpressing FGF21 exhibit a range of metabolism-related phenotypes, including slow growth rates, low blood glucose and triglyceride levels, resistance to age-related type 2 diabetes, islet hyperplasia, and obesity. In rat or non-human primate models, animals show changes such as normal recovery of blood glucose levels, decreased blood triglyceride and cholesterol levels, and increased glucose tolerance and insulin sensitivity after injection of FGF21 fusion protein. Simultaneously, FGF21 increases animal activity, metabolic rate, and energy expenditure, and further reduces animal body weight and body fat. In a mouse model of non-alcoholic steatohepatitis, modified FGF21 protein improves the degree of hepatic steatosis, reduces liver damage, and decreases molecular markers of inflammation in the liver. These studies suggest that FGF21 may be useful in treating metabolism-related diseases such as non-alcoholic steatohepatitis, type 2 diabetes, obesity, and dyslipidemia.

[0006] Of the FGF receptors, only FGFR1c, 2c, 3c, and 4 can bind to Klotho, while FGFR1b, 2b, and 3b cannot. In vitro experiments have shown that FGF21 activates FGFR1c, 2c, and 3c receptors with the help of βKlotho. In vivo experiments have shown that the metabolic regulatory effects of FGF21 are mainly mediated by FGFR1c and βKlotho receptors, and that knocking out FGFR1c or βKlotho significantly weakens the metabolic regulating effects of FGF21. Therefore, it is hypothesized that molecules that mimic the action of FGF21 and can activate FGFR1c and βKlotho receptors (e.g., activated βKlotho antibodies) may be useful in treating metabolic diseases such as non-alcoholic steatohepatitis.

[0007] Currently, there is limited research on antibodies targeting β-Klotho. Relatively rapid progress is seen in Genentech's BFKB8488A (anti-FGFR1 / KLB bispecific antibody) in Phase II clinical trials, and NGM Biopharmaceuticals' NGM313 (monoclonal stimulating antibody) also in Phase II clinical trials (all clinical trials targeting non-alcoholic steatohepatitis). Other companies, such as Amgen, Regeneron, and Novartis, have research products in the preclinical stage. Because all of these drugs are in the early stages of clinical trials and their therapeutic effects are not yet established, there is a market demand for antibody drugs targeting these substances for a wider range of therapies. [Overview of the project]

[0008] According to a first aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ ID The sequence is either the sequence indicated by NO. 5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 5, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 15, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 15, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0009] (3) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.20; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 21, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 21, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 26, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 26; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in NO.4, the sequence of LCDR2 is the sequence shown in SEQ ID NO.27, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in SEQ ID NO.27, the sequence of LCDR3 is the sequence shown in SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in SEQ ID NO.6, or

[0010] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.31; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 5, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 34, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 34, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0011] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 27, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 27, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 42, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0012] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.20; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 45, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 45, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 31; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or SEQ ID The sequence indicated by NO.4 has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.42, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6, Here, the sequences of HCDR and LCDR provide an anti-βKlotho antibody or its antigen-binding moiety, as defined by Kabat.

[0013] According to a second aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 10; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 11, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 16, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 16; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0014] (3) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 22, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 22; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 28, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 28; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0015] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 35, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 35; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0016] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 39, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 39; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0017] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 46, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 46; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence indicated by NO.10 has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6, Here, the sequences HCDR and LCDR provide an anti-βKlotho antibody or its antigen-binding moiety, as defined by IMGT.

[0018] According to a third aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, The heavy-chain variable region has an amino acid sequence represented by SEQ ID NO. 12, 18, 24, 29, 33, 36, 41, 43, 47 or 49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the amino acid sequence represented by SEQ ID NO. 12, 18, 24, 29, 33, 36, 41, 43, 47 or 49, and / or The light-chain variable region has an amino acid sequence represented by SEQ ID NO. 13, 19, 25, 30, 37, 44 or 48, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the amino acid sequence represented by SEQ ID NO. 13, 19, 25, 30, 37, 44 or 48, Provided is an anti-βKlotho antibody or an antigen-binding portion thereof.

[0019] According to a fourth aspect, the present invention is an anti-βKlotho antibody or an antigen-binding portion thereof comprising a heavy-chain variable region, wherein the heavy-chain variable region comprises heavy-chain CDR1 (HCDR1), heavy-chain CDR2 (HCDR2), and heavy-chain CDR3 (HCDR3), (1) The sequence of HCDR1 is the sequence represented by SEQ ID NO. 1, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence represented by SEQ ID NO. 1, the sequence of HCDR₂ is the sequence represented by SEQ ID NO. 2, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence represented by SEQ ID NO. 2, and the sequence of HCDR3 is the sequence represented by SEQ ID NO. 3, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence represented by SEQ ID NO. 3, or (2) The sequence of HCDR1 is the sequence shown in SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.1; the sequence of HCDR2 is the sequence shown in SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.14; the sequence of HCDR3 is the sequence shown in SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.3; or

[0020] (3) The sequence of HCDR1 is the sequence shown in SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.1; the sequence of HCDR2 is the sequence shown in SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.20; the sequence of HCDR3 is the sequence shown in SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the sequence shown in SEQ ID NO.3; or (4) The sequence of HCDR1 is the sequence shown in SEQ ID NO. 26, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 26; the sequence of HCDR2 is the sequence shown in SEQ ID NO. 14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 14; the sequence of HCDR3 is the sequence shown in SEQ ID NO. 3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 3;

[0021] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.31; (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3;

[0022] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 31; Here, the HCDR sequence provides an anti-β-Klotho antibody or its antigen-binding moiety, as defined by Kabat.

[0023] According to a fifth aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a light chain variable region, wherein the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO.5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.5; the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6; (2) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 15, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 15; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6;

[0024] (3) The sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO.21, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.21; the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6; (4) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 27, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 27; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; (5) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 34, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 34; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6;

[0025] (6) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 42; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; (7) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 45, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 45; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; Here, the LCDR sequence provides an anti-β-Klotho antibody or its antigen-binding moiety, as defined by Kabat.

[0026] According to a sixth aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 16, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 16; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0027] (3) The sequence of HCDR1 is the sequence shown in SEQ ID NO. 22, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 22; the sequence of HCDR2 is the sequence shown in SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 23; the sequence of HCDR3 is the sequence shown in SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 9; (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 28, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 28; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0028] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 35, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 35; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 39, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 39; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0029] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 46, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 46; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; Here, the HCDR sequence, defined by IMGT, provides an anti-βKlotho antibody or its antigen-binding moiety.

[0030] According to a seventh aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a light chain variable region, wherein the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 10; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 11; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; Here, the LCDR sequence provides an anti-βKlotho antibody or its antigen-binding moiety, as defined by IMGT.

[0031] According to the eighth aspect, the present invention provides an isolated nucleic acid molecule encoding an anti-βKlotho antibody or its antigen-binding moiety as described in any of the first to seventh aspects. According to the ninth aspect, the present invention provides a vector containing the nucleic acid molecule described in the eighth aspect. According to the tenth aspect, the present invention provides a host cell containing a nucleic acid molecule as described in the eighth aspect or a vector as described in the ninth aspect.

[0032] According to the eleventh aspect, the present invention provides an antibody-drug conjugate comprising an anti-β-Klotho antibody or its antigen-binding moiety according to any embodiment of the first to seventh aspects, which is conjugated to a therapeutic agent. According to the twelfth aspect, the present invention provides a pharmaceutical composition comprising an anti-βKlotho antibody or its antigen-binding moiety according to any of the first to seventh aspects, or an antibody-drug conjugate according to the eleventh aspect, and a pharmaceutically acceptable carrier.

[0033] According to the 13th aspect, the present invention relates to the use in the manufacture of a pharmacopoeia for the treatment of a disease mediated by βKlotho or βKlotho-FGFR1c-FGF21, of an anti-βKlotho antibody or antigen-binding moiety thereof according to any of the 1st to 7th aspects, a nucleic acid molecule according to the 8th aspect, a vector according to the 9th aspect, a host cell according to the 10th aspect, or an antibody-drug conjugate according to the 11th aspect, wherein the disease is, for example, a metabolic disorder, such as type 1 diabetes mellitus, type 2 diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

[0034] According to the 14th aspect, the present invention provides a method for treating a disease in an individual that is mediated by βKlotho or βKlotho-FGFR1c-FGF21, comprising administering to the individual a therapeutically effective amount of an anti-βKlotho antibody or its antigen-binding moiety according to any of the 1st to 7th aspects, an antibody-drug conjugate according to the 11th aspect, or a pharmaceutical composition according to the 12th aspect, wherein the disease is, for example, a metabolic disorder, such as type 1 diabetes mellitus, type 2 diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

[0035] According to the 15th aspect, the present invention provides a conjugate comprising an anti-βKlotho antibody or its antigen-binding moiety as described in any of the 1st to 7th aspects, and a detectable marker. According to the sixteenth aspect, the present invention provides a fusion protein comprising an anti-βKlotho antibody or its antigen-binding moiety as described in any of the first to seventh aspects. [Brief explanation of the drawing]

[0036] [Figure 1] This figure shows the concentration profiles of the human or crab-eating monkey FGFR1c / βKlotho receptor upon activation by the antibody CB-8-42, a control antibody, and FGF19, as determined by a reporter gene assay. [Figure 2] This figure shows the results of monitoring the body weight of crab-eating monkeys treated with a solvent or the antibody CB-8-42. [Figure 3] This figure shows the results of monitoring relative body weight changes in crab-eating monkeys treated with a solvent or the antibody CB-8-42. [Figure 4] This figure shows the monitoring results of BMI in crab-eating monkeys treated with a solvent or the antibody CB-8-42.

[0037] [Figure 5] This figure shows the results of monitoring the food intake of crab-eating monkeys treated with a solvent or the antibody CB-8-42. [Figure 6] This figure shows the change in liver fat content in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Figure 7] This figure shows the changes in fasting blood glucose in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Figure 8] This figure shows the changes in serum insulin levels in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42.

[0038] [Figure 9] This figure shows the changes in serum triglyceride levels in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Figure 10] This figure shows the changes in serum total cholesterol levels in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Figure 11] This figure shows the changes in serum low-density lipoprotein levels in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Figure 12] This figure shows the changes in serum high-density lipoprotein levels in crab-eating monkeys before and after treatment with a solvent or the antibody CB-8-42. [Modes for carrying out the invention]

[0039] [Detailed description of the invention] [Definition] Unless otherwise specified, all scientific and technical terms used herein have the same meaning as those understood by those skilled in the art. For definitions and terminology in this art, those skilled in the art can refer in particular to *Current Protocols in Molecular Biology* (Ausubel). Amino acid residue abbreviations are standard three-letter and / or one-letter codes used in this art, referring to one of 20 common L-amino acids.

[0040] Regardless of the numerical ranges and parameter approximations shown in the broader scope of this invention, the numerical values ​​shown in the specific examples are described as accurately as possible. However, all numerical values ​​inherently contain a certain degree of error due to the standard deviation present in each measurement. Furthermore, all ranges disclosed herein should be understood as covering all subranges included within that range. For example, the described range "1 to 10" should be considered to include any and all subranges between the minimum value of 1 and the maximum value of 10 (including the endpoints), i.e., all subranges starting from a minimum value of 1 or greater, e.g., 1 to 6.1, and all subranges ending at a maximum value of 10 or less, e.g., 5.5 to 10. Also, any references referred to as "incorporated herein" should be understood as incorporated as a whole.

[0041] As used herein, the terms “subject” or “individual” mean a mammal such as a human, but may also mean other animals such as wild animals, domesticated animals, or laboratory animals (e.g., orangutans, monkeys, rats, mice, rabbits, guinea pigs, tarbagans, ground squirrels, etc.). As used herein, the term “antigen” refers to a predetermined target to which an antibody can selectively bind. Examples of antigens include, but are not limited to, polypeptides, sugars, nucleic acids, lipids, haptens, or other natural or synthetic compounds.

[0042] In a broad sense, "antibody" can refer to an immunoglobulin molecule that can specifically bind to a target via at least one antigen-recognition site located in the variable region of the immunoglobulin molecule, and therefore encompasses complete antibodies / full-length antibodies, single-chain antibodies, or any antigen-binding fragment of an antibody (also called the "antigen-binding moiety"). When "antibody" and "antigen-binding fragment / antigen-binding moiety" appear in the same context, "antibody" can be understood as the complete form of "antigen-binding fragment / antigen-binding moiety," and both commonly correspond to the broad concept of antibody.

[0043] As used herein, the terms “βklotho” or “βklotho polypeptide” and similar terms include polypeptides derived from any vertebrate, including mammals such as primates (e.g., humans, cynos), dogs, and rodents (e.g., mice and rats) (the terms “polypeptide” and “protein” are used interchangeably herein), or any naturally occurring βklotho, and unless otherwise noted, in some embodiments, include the relevant βklotho polypeptide and its SNP variants. βklotho comprises two domains: βklotho1 (KLB1) and βklotho2 (KLB2). Each βklotho domain contains a glycosylhydrolase 1 region. For example, the KLB1 domain of human β-klotho contains amino acid residues 1-508, and its glycosyl hydrolase 1 region contains amino acid residues 77-508. The KLB2 domain of human β-klotho contains amino acid residues 509-1044, and its glycosyl hydrolase 1 region contains amino acid residues 517-967. The amino acid sequence of human β-klotho (NCBI Reference Sequence: NM_175737.4) is as follows.

[0044]

[0045] βklotho polypeptides further include allele variants (e.g., SNP variants), splicing variants, fragments, derivatives, substitution / deletion and insertion variants, fusion polypeptides and interspecies homologues sufficient to maintain βklotho activity and / or generate an anti-βklotho immune response. Those skilled in the art should recognize that the anti-βklotho antibodies provided herein may bind to βklotho polypeptides, βklotho polypeptide fragments, βklotho antigens and / or βklotho epitopes. An epitope may be part of a larger βklotho antigen, part of a larger βklotho polypeptide fragment, or part of a larger βklotho polypeptide. βklotho can exist in native or modified forms. The βklotho polypeptides described herein can be isolated from various sources, e.g., human tissue-type sources or other sources, or prepared by recombinant or synthetic methods. βklotho polypeptides may include polypeptides having the same amino acid sequence as the corresponding naturally occurring βklotho polypeptide. βklotho polypeptides include cleaved or secreted forms of βklotho polypeptide (e.g., extracellular domain sequences), mutant forms of the polypeptide (e.g., alternative splicing forms), and allelic variants. Orthologs of βklotho polypeptide are also well known in the art.

[0046] The term "fibroblast growth factor" refers to a family of growth factors that include 22 members of the human FGF family. The FGF19 subfamily of fibroblast growth factors consists of human FGF21, FGF23, and FGF19, as well as mouse FGF15. The role of each member of the FGF family is the result of heparin-dependent binding to one or more members of the FGF receptor tyrosine kinase (FGFR) family, which includes four members, each possessing a tyrosine kinase domain: FGFR1, FGFR2, FGFR3, and FGFR4, as well as two splicing variants of each of FGFR1, FGFR2, and FGFR3. These splicing variants occurring in exon 3 of FGFR1, FGFR2, and FGFR3 are named "b" and "c" variants (for example, FGFR1b, FGFR2b, FGFR3c, FGFR1c, FGFR2c, and FGFR3c, which are also called FGFR1(III)b, FGFR2(III)b, FGFR3(III)b, FGFR1(III)c, FGFR2(III)c, and FGFR3(III)c, respectively).

[0047] The terms “anti-βKlotho antibody” or “antibody that binds to βklotho” include antibodies that can bind to βklotho with sufficient affinity so that the antibody can be used as a diagnostic and / or therapeutic agent when targeting βklotho. Preferably, the anti-βKlotho antibody binds to unrelated non-βklotho proteins to an extent of less than about 10% of the antibody binding to the βklotho protein as measured, for example, by fluorescence-activated cell sorting (FACS) analysis or immunoassay, such as radioimmunoassay (RIA). As described above, antibodies that “specifically bind” to βklotho, or antibodies that “specific” to βklotho, have been explained. In some embodiments, the dissociation constant (Kd) of the antibodies described herein that bind to βklotho is 500 nM, 100 nM, 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM or less. In some embodiments, the anti-βKlotho antibody binds to an epitope of βklotho that is conserved between βklotho from different species (e.g., between human and crab-eating monkey βklotho).

[0048] The term "agonist antibody" refers to an antibody that induces a response, such as an antibody that mimics at least one functional activity of a target polypeptide (e.g., FGF19 or FGF21). Agonist antibodies include ligand mimics, where, for example, the ligand binds to a cell surface receptor, and this binding induces cell signaling or activity via an intracellular signaling pathway, and the antibody induces similar cell signaling or activation.

[0049] A "full-length antibody (intact antibody)" refers to a protein containing at least two heavy chains (H) and two light chains (L) linked to each other via disulfide bonds. Each heavy chain contains a heavy chain variable region (abbreviated as VH) and a heavy chain constant region. The heavy chain constant region contains three domains: CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated as VL) and a light chain constant region. The light chain constant region contains domain CL. The VH and VL regions may be further subdivided into multiple hypervariable regions called complementarity-determining regions (CDRs), which contain multiple more conservative regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. These variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant domain of an antibody can mediate the binding of immunoglobulins to various cells of the immune system (such as effector cells) and to host tissues or factors, including the first component (Clq) of the classical complement system. Full-length antibodies (intact antibodies) may be any type of antibody, such as IgD, IgE, IgG, IgA, or IgM (or any of the aforementioned subclasses), but the antibody does not need to belong to any particular class. Immunoglobulins can be designated into different classes based on the antibody amino acid sequence of the constant domain of the heavy chain. Typically, there are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which can be further classified into subclasses (isotypes) such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different immunoglobulin classes are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional structures of different classes of immunoglobulins are known. Chimeric antibodies or humanized antibodies are also included in the antibodies according to the present invention. Those skilled in the art know that the complementarity-determining regions (CDRs, usually CDR1, CDR2, and CDR3) are the regions within the variable region that have the greatest influence on the affinity and specificity of the antibody.The CDR amino acid sequences of the VH or VL regions are defined using many common definition methods, such as the IMGT definition, the Chothia definition, and the Kabat definition. For a given antibody's variable region amino acid sequence, the CDR amino acid sequences in the VH and VL amino acid sequences can usually be determined by various definition methods. In embodiments of the present invention, the CDR amino acid sequence is defined using Kabat or IMGT. For a given antibody's variable region amino acid sequence, the CDR amino acid sequence of the variable region amino acid sequence can be analyzed using various methods.

[0050] The term "mouse antibody" refers to antibodies obtained by screening mouse cross fusion cells—created by fusing B cells and myeloma cells of immunized mice—which can proliferate indefinitely and secrete antibodies, followed by further screening, manufacturing, and purification. Mouse antibodies generally possess immunogenicity, and therefore require subsequent humanization treatment.

[0051] The term "humanized antibody" refers to an antibody obtained by transplanting a CDR sequence derived from another mammalian species, such as a mouse, onto a human framework sequence. Several residues in the backbone (called FR) section can be modified to maintain binding affinity. Humanized antibodies or fragments thereof according to the present invention can be manufactured by techniques well known to those skilled in the art.

[0052] The term "chimeric antibody" refers to an antibody in which the variable region sequence originates from one species and the constant region sequence originates from another species, for example, an antibody in which the variable region sequence originates from a mouse antibody and the constant region sequence originates from a human antibody. The chimeric antibody or fragment thereof according to the present invention can be produced using genetic recombination technology. For example, the chimeric antibody can be produced by cloning recombinant DNA containing a promoter, a sequence encoding the variable region of a non-human (particularly mouse) monoclonal antibody described in the present invention, and a sequence encoding the constant region of a human antibody. The chimeric antibody according to the present invention encoded by such a recombinant gene is, for example, a mouse-human chimera, and the specificity of the antibody is determined by the variable region derived from mouse DNA, and its isotype is determined by the constant region derived from human DNA.

[0053] The term "partially humanized antibody" refers to an antibody that contains a constant region derived from humans and a variable region (including the CDR) derived from non-human organisms (such as mice). The term "semi-humanized antibody" refers to a type of humanized antibody in which one antibody chain contains the mouse variable region and the other antibody chain contains the humanized variable region; in other words, a semi-humanized antibody. The term "monoclonal antibody" refers to an antibody obtained from a nearly identical antibody population (where the individual antibodies constituting the population are identical except for the possibility of spontaneous mutations occurring in a small number of individuals).

[0054] As used herein, the terms “antigen-binding fragment,” “antigen-binding portion,” or “antigen-binding region” refer to a portion of an antibody that is interchangeable, interacts with an antigen, and confers specificity and affinity to the antigen to the binder, particularly antibody fragments such as Fv, Fab, F(ab')2, or Fab', or any fragment whose half-life can be extended by chemical modification or incorporation into a liposome, wherein the chemical modification is, for example, the addition of a poly(alkylene) glycol such as polyethylene glycol ("polyethylene glycolization, PEGylation") (PEGylated fragments of Fv-PEG, scFv-PEG, Fab-PEG, F(ab')2-PEG, or Fab'-PEG) ("PEG": polyethylene glycol), and the fragment has β-Klotho-binding activity. Preferably, the functional fragment consists of or includes a sub-sequence of the variable chain of the heavy or light chain of the derived antibody, the sub-sequence being sufficient to maintain the same binding specificity and sufficient affinity as the derived antibody, and the functional fragment includes at least five amino acids, preferably 10, 15, 25, 50, and 100 consecutive amino acids of the derived antibody sequence. Examples of antigen-binding fragments include, but are not limited to, (1) a Fab fragment which may be a monovalent fragment having a VL-CL chain and a VH-CH1 chain, (2) a F(ab')2 fragment which may be a bivalent fragment having two Fab' fragments linked by a disulfide bridge in the hinge region (i.e., a dimer of Fab'), and (3) an Fv fragment with a VL domain and a VH domain having a single arm of the antibody.

[0055] The term "single-chain antibody (scFv)" refers to a single polypeptide chain in which VH domains and VL domains are linked via a peptide linker. (scFv)2 contains two VH domains linked via a peptide linker and two VL domains linked to these two VH domains via disulfide crosslinks.

[0056] The terms "Fc fragment," "Fc region," "Fc domain," and "Fc portion," or similar terms, refer to a portion of the constant region of the antibody heavy chain, including the hinge region and the CH2 and CH3 fragments of the constant region. The Fc region of anti-βKlotho antibodies can be engineered or modified, for example, to reduce or eliminate antibody-dependent cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), including modifications related to effector function, which can be achieved by introducing one or more amino acid substitutions / mutations into the antibody's Fc region.

[0057] As used herein, the term "specific binding" refers to, for example, the binding of an antibody to an antigen epitope, or a non-random binding reaction between two molecules. The term "polyantibody," also known as a "multispecific antibody," refers to a molecule that has binding specificity to at least two different antigens. Such molecules that bind to only two antigens are also called biantibodies (i.e., bispecific antibodies, BsAbs).

[0058] The term "bispecific antibody" refers to an antibody that has the ability to simultaneously bind to two different antigenic epitopes. The two antigenic epitopes may be located on different antigens or on the same antigen. Bispecific antibodies may have multiple structural configurations. For example, a bispecific antibody may consist of two Fc fragments and two antigen-binding moieties fused to each of them (similar to a native antibody except that the two arms bind to different antigenic targets or epitopes), and the antigen-binding moieties may be in the form of a single-chain antibody (scfv) or a Fab fragment.

[0059] Typically, to prepare monoclonal antibodies or their functional fragments, particularly mouse-derived monoclonal antibodies or their functional fragments, one can refer to the techniques described in the manual "Antibodies" (Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor NY, pp. 726, 1988), or the technique by Kohler and Milstein for production from hybridoma cells (Nature, 256:495-497, 1975).

[0060] The term “conservative variant” or “conservative amino acid substitution” refers to a substitution that does not substantially affect or reduce the affinity of a protein, for example, the affinity of an antibody to βKlotho. For example, a human antibody that specifically binds to βKlotho may contain approximately 1 or fewer, approximately 2 or fewer, approximately 5 or fewer, approximately 10 or fewer, or approximately 15 or fewer conservative substitutions and still specifically binds to the βKlotho polypeptide. The term “conservative variant” also includes using a substituted amino acid instead of an unsubstituted parent amino acid, as long as the antibody specifically binds to βKlotho. Non-conservative substitutions reduce activity or bind to βKlotho.

[0061] The term “isolated” biological component (e.g., nucleic acid, protein (including antibodies), or organelle) refers to a component that has been substantially isolated or purified from other biological components (i.e., other chromosomes and additional chromosomal DNA or RNA, proteins, and organelles) in its naturally occurring environment (e.g., cells). Already “isolated” nucleic acids and proteins include those purified by standard purification methods. This term also includes nucleic acids and proteins produced by recombinant expression in host cells, as well as chemically synthesized nucleic acids.

[0062] The term "inducing sequence" refers to a sequence that has at least 80% (preferably 85%, 90%, 95%, 98%, or 99%) sequence identity with the related sequence and has the same or similar function.

[0063] As used herein, the term “pharmaceutical composition” refers to a combination of at least one drug and a pharmaceutically usable carrier or auxiliary substance, which are combined together to achieve a particular purpose. In some embodiments, the pharmaceutical composition includes combinations that are temporally and / or spatially separated, insofar as they can act together to achieve the purpose of the present invention. For example, the components contained in the pharmaceutical composition (e.g., antibodies, nucleic acid molecules, combinations of nucleic acid molecules, and / or conjugates according to the present invention) may be administered to an individual as a whole or in separate doses. When the components contained in the pharmaceutical composition are administered to an individual in separate doses, the components may be administered to the individual simultaneously or sequentially. Preferably, the pharmaceutical carrier is water, a buffered aqueous solution, an isotonic salt solution such as PBS (phosphate buffer), glucose, mannitol, dextrorotatory glucose, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerin, hyaluronic acid, ethanol, or polyalkylene glycols such as polypropylene glycol and triglycerides. The type of pharmaceutical carrier used depends, in particular, on whether the composition according to the present invention is formulated for oral, nasal, intradermal, subcutaneous, intramuscular, or intravenous administration. The composition according to the present invention may contain humectants, emulsifiers, or buffering agents as additives. The pharmaceutical composition or pharmaceutical formulation according to the present invention may be administered by any suitable route, such as oral, nasal, intradermal, subcutaneous, intramuscular, or intravenous administration.

[0064] As used herein, the terms “therapeutic effective dose” or “effective dose” refer to a dose sufficient to demonstrate its benefit to the individual being administered. The actual amount administered, the rate of administration, and the course of time will depend on the condition and severity of the individual being treated. The prescription for treatment (e.g., dosage determination) is ultimately the responsibility of the specialist and other physicians and is generally determined by considering the disease being treated, the individual patient’s condition, the site of delivery, the method of administration, and other factors known to the physician.

[0065] EC 50 The value primarily refers to the concentration of a corresponding drug, antibody, or toxin that can reach 50% of its maximum biological effect after a specific exposure time. In pharmacology, it is used to characterize the activating ability of an agonist in in vitro experiments, and also to indicate the blood concentration required to reach half of the maximum biological effect in vivo. In some literature, the EC 50 It is also used to characterize the potency of a compound at a certain cellular level (including agonism and antagonistism), and EC 50 The value can be measured by methods such as ELISA.

[0066] As used herein, the term "fusion protein" generally refers to a protein consisting of at least two domains, each domain being unrelated in its natural state, encoded by individual genes, and where these genes are linked together and transcribed and translated as a whole to produce a single protein. In the technical background of the present invention, a "fusion protein" containing an antibody or antigen-binding moiety refers to a product obtained by fusing an antibody or antigen-binding moiety with another bioactive protein using genetic engineering techniques. Such an antibody fusion protein combines the antigen-binding ability of the antibody with the unique biological properties of the bioactive protein fused with the antibody.

[0067] The term "identity / homology / consistency" of an amino acid or nucleic acid sequence is defined as the proportion of identical residues in an amino acid or nucleotide sequence variant after sequence alignment and capping, to the maximum proportion of identity as required. Methods and computer programs for alignment are known to those skilled in the art.

[0068] According to a first aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ ID The sequence is either the sequence indicated by NO. 5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 5, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 15, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 15, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0069] (3) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.20; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 21, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 21, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 26, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 26; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in NO.4, the sequence of LCDR2 is the sequence shown in SEQ ID NO.27, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in SEQ ID NO.27, the sequence of LCDR3 is the sequence shown in SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence shown in SEQ ID NO.6, or

[0070] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.31; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 5, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 34, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 34, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0071] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 27, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 27, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 42, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or

[0072] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.20; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; the sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 45, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 45, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 31; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or SEQ ID The sequence indicated by NO.4 has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.42, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6, Here, the sequences of HCDR and LCDR provide an anti-βKlotho antibody or its antigen-binding moiety, as defined by Kabat.

[0073] According to a second aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region comprises light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 10; the sequence of LCDR2 is SEQ The sequence is either the sequence indicated by ID NO. 11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 11, and the sequence of LCDR3 is either the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6, or (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 16, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 16; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0074] (3) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 22, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 22; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 28, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 28; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0075] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 35, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 35; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0076] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 39, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 39; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or

[0077] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 46, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 46; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence of LCDR2 is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by NO.10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence indicated by SEQ ID NO.6, or (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or SEQ ID The sequence indicated by NO.10 has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity, the sequence of LCDR2 is the sequence indicated by SEQ ID NO.11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.11, the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6, Here, the sequences HCDR and LCDR provide an anti-βKlotho antibody or its antigen-binding moiety, as defined by IMGT.

[0078] In some embodiments of the first and second aspects, the heavy chain variable region has an amino acid sequence represented by SEQ ID NO. 12, 18, 24, 29, 33, 36, 41, 43, 47, or 49, or has an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence represented by SEQ ID NO. 12, 18, 24, 29, 33, 36, 41, 43, 47, or 49.

[0079] In some embodiments of the first and second aspects, the light chain variable region has an amino acid sequence represented by SEQ ID NO. 13, 19, 25, 30, 37, 44, or 48, or has an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence represented by SEQ ID NO. 13, 19, 25, 30, 37, 44, or 48.

[0080] In some embodiments of the first and second aspects, (1) the heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 12, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 13, or (2) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 18, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 19, or (3) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 24, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 25, or (4) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 29, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 30, or

[0081] (5) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 33, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 13, or (6) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 36, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 37, or (7) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 41, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 30, or

[0082] (8) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 43, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 44, or (9) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 47, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 48, or (10) The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 49, and the light chain variable region sequence is the sequence indicated by SEQ ID NO. 44.

[0083] According to a third aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, The heavy chain variable region has an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence represented by SEQ ID NO. 12, 18, 24, 29, 33, 36, 41, 43, 47, or 49, and / or The light chain variable region has an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence that is at least 80%, at least 95%, or at least 99% identical to the amino acid sequence that is5%, at least 90%, at least 95%, or at least 99% identical to the amino acid sequence that is at least 90%, at least 85%, at least 90%, at least 95%, This provides an anti-βKlotho antibody or its antigen-binding moiety.

[0084] According to a fourth aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.14; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3;

[0085] (3) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.20, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.20; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3; (4) The sequence of HCDR1 is the sequence shown in SEQ ID NO. 26, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 26; the sequence of HCDR2 is the sequence shown in SEQ ID NO. 14, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 14; the sequence of HCDR3 is the sequence shown in SEQ ID NO. 3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 3;

[0086] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.2, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.2; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.31; (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO.1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO.38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO.3, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.3;

[0087] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 1, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 1; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 38, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 38; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 31, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 31; Here, the HCDR sequence provides an anti-β-Klotho antibody or its antigen-binding moiety, as defined by Kabat.

[0088] According to a fifth aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a light chain variable region, The light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), (1) The sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO.5, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.5; the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6; (2) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 15, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 15; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6;

[0089] (3) The sequence of LCDR1 is the sequence indicated by SEQ ID NO.4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO.21, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.21; the sequence of LCDR3 is the sequence indicated by SEQ ID NO.6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO.6; (4) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 27, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 27; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6;

[0090] (5) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 34, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 34; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; (6) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 42, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 42; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6;

[0091] (7) The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 4; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 45, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 45; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; Here, the LCDR sequence provides an anti-β-Klotho antibody or its antigen-binding moiety, as defined by Kabat.

[0092] According to a sixth aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region, wherein the heavy chain variable region comprises heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), (1) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (2) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 16, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 16; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0093] (3) The sequence of HCDR1 is the sequence shown in SEQ ID NO. 22, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 22; the sequence of HCDR2 is the sequence shown in SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 23; the sequence of HCDR3 is the sequence shown in SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence shown in SEQ ID NO. 9; (4) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 28, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 28; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0094] (5) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 8, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 8; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; (6) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 35, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 35; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 17, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 17; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0095] (7) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 39, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 39; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (8) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9;

[0096] (9) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 46, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 46; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 23, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 23; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 9, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 9; (10) The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 7; the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 40; the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 32; Here, the HCDR sequence, defined by IMGT, provides an anti-βKlotho antibody or its antigen-binding moiety.

[0097] According to a seventh aspect, the present invention relates to an anti-βKlotho antibody or its antigen-binding moiety comprising a light chain variable region, The light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), The sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 10; the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 11, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 11; the sequence of LCDR3 is the sequence indicated by SEQ ID NO. 6, or has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the sequence indicated by SEQ ID NO. 6; Here, the LCDR sequence provides an anti-βKlotho antibody or its antigen-binding moiety, as defined by IMGT.

[0098] In some embodiments of the first to seventh aspects, the anti-βKlotho antibody or its antigen-binding moiety binds to human βKlotho. In some specific embodiments, the anti-βKlotho antibody or its antigen-binding moiety specifically binds to human βKlotho. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody is a complete antibody, a single-chain antibody (scFv), a bispecific antibody, or a multispecific antibody. In some embodiments of the first to seventh aspects, the antigen-binding moiety of the anti-βKlotho antibody is Fab, Fab', Fv, or F(ab')2.

[0099] In some embodiments of the first to seventh aspects, the anti-βKlotho antibody is a mouse antibody, a chimeric antibody, a humanized antibody, or a fully human antibody. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody is a monoclonal antibody. In some embodiments of the first to seventh aspects, the anti-Klotho antibody is an IgM, IgD, IgG, IgA, or IgE type antibody. In some specific embodiments, the anti-Klotho antibody is an IgG antibody. In some more specific embodiments, the anti-Klotho antibody is an IgG1 antibody.

[0100] In some embodiments of the first to seventh aspects, the anti-βKlotho antibody is an IgG1, IgG2, or IgG4 isotype. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody is an IgG1 / IgG4, IgG2 / IgG4, or IgG1 / IgG2 chimeric type. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody comprises a constant light chain region of the κ subtype or the λ subtype.

[0101] In some embodiments of the first to seventh aspects, the anti-βKlotho antibody comprises a human IgG1 heavy chain constant region and a human κ light chain constant region. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody or its antigen-binding moiety is used for medical treatment. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody or its antigen-binding moiety has ADCC activity or CDC activity.

[0102] In some embodiments of the first to seventh aspects, the anti-βKlotho antibody includes a wild-type Fc region. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody or its antigen-binding moiety has enhanced ADCC activity or CDC activity through engineering modifications. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody or its antigen-binding moiety has reduced ADCC activity or CDC activity due to engineering modifications.

[0103] For the in vivo application of β-Klotho and antibodies, which are the targets of this invention, antibodies with weakened or even absent ADCC or CDC activity may be advantageous, because an overly strong Fc effect can affect, and even weaken, the efficacy of the antibody. Means for weakening or eliminating the Fc effect are known in the art and include, for example, 1) antibody modification of amino acid residues (mainly weakening binding to the relevant receptor), 2) glycosylation modification, and 3) IgG4 antibody modification. In some embodiments of the first to seventh aspects, the anti-βKlotho antibody has a heavy chain variable region indicated by SEQ ID NO. 49, a light chain variable region indicated by SEQ ID NO. 44, a heavy chain constant region indicated by SEQ ID NO. 50, and a light chain constant region indicated by SEQ ID NO. 51.

[0104] According to the eighth aspect, the present invention provides an isolated nucleic acid molecule encoding an anti-βKlotho antibody or its antigen-binding moiety as described in any of the first to seventh aspects. In some embodiments of the first to fifth aspects, the antibody or its antigen-binding moiety according to the present invention may be a chimeric antibody or a humanized antibody, the humanized antibody including a semi-humanized antibody, a partially humanized antibody or a fully human antibody. In some embodiments, the polynucleotide is operably linked to a regulatory sequence, the regulatory sequence may be recognized by host cells transformed with the vector.

[0105] According to the ninth aspect, the present invention provides a vector (e.g., an expression vector) comprising a combination of nucleic acid molecules or polynucleotides as described in the eighth aspect. In some embodiments, the expression vector according to the present invention comprises a nucleic acid molecule or a combination of polynucleotides described in the present invention, wherein the polynucleotide is effectively ligated to a regulatory sequence that enables the polypeptide encoded thereby to be expressed in a host cell or a cell-free expression system. The selection of the expression vector is by choice of host cell and may be selected to have desired expression and regulatory characteristics in the chosen host cell.

[0106] An "expression vector" is a vector containing one or more expression regulatory sequences, and an "expression regulatory sequence" is a DNA sequence that controls and regulates the transcription and / or translation of other DNA sequences. Nucleic acids in a vector may be operably ligated to one or more expression regulatory sequences. As used herein, “operably ligated” means that the expression regulatory sequence is incorporated into the genetic construct so as to effectively control the expression of the coding sequence of interest. Examples of expression regulatory sequences include promoters, enhancers, and transcription termination regions. A promoter is an expression regulatory sequence consisting of a region of a DNA molecule located 100 nucleotides upstream of the transcription start site (typically near the RNA polymerase II start site). To place the coding sequence under the control of a promoter, the translation start site of the polypeptide translation reading frame must be positioned between 1 and approximately 50 nucleotides downstream of the promoter. Enhancers provide temporal, spatial, and level expression specificity. Unlike promoters, enhancers can function at different distances from the transcription site. Enhancers may be located downstream of the transcription start site. If RNA polymerase can transcribe the coding sequence into mRNA, and the mRNA can then be translated into the protein encoded by the coding sequence, the coding sequence is "operably ligated" to an intracellular regulatory expression sequence and is "under the control" of that regulatory expression sequence.

[0107] Suitable expression vectors include, but are not limited to, plasmids and viral vectors derived from phages, baculoviruses, tobacco mosaic viruses, herpesviruses, cytomegaloviruses, retroviruses, vaccinia viruses, adenoviruses, and adeno-associated viruses. Many vectors and expression systems are commercially available from companies such as Novagen (Madison, WI), Clontech (Palo Alto, CA), Stratagene (La Jolla, CA), and Invitrogen Life Technologies (Carlsbad, CA).

[0108] The expression vector may include a tag sequence. The tag sequence is typically expressed as a fusion with the encoded polypeptide. Such a tag may be inserted at any position within the polypeptide, including carboxyl or amino-terminants. Examples of useful tags include, but are not limited to, Fc fragments, polyhistidine, green fluorescent protein (GFP), glutathione S-transferase (GST), c-myc, hemagglutinin, Flag™ tag (Kodak, NewHaven, CT), maltose E-binding protein, and protein A. In some embodiments, the nucleic acid molecule encoding the βKlotho fusion polypeptide resides in a vector containing a nucleic acid encoding one or more domains of the Ig heavy chain constant region, where the domains correspond, for example, to the amino acid sequences (Fc fragments) of the hinge region, CH2 region, and CH3 region of the human immunoglobulin Cγ1 chain.

[0109] According to the tenth aspect, the present invention provides a host cell containing a nucleic acid molecule as described in the eighth aspect or a vector as described in the ninth aspect. In some embodiments, the host cell may be a prokaryotic host cell, a eukaryotic host cell, or a phage. The prokaryotic host cell may be Escherichia coli, Bacillus subtilis, Streptomyces, or Proteus mirabilis, etc. The eukaryotic host cell may be a fungus such as Pichia pastoris, budding yeast (Saccharomyces cerevisiae), fission yeast (Schizosaccharomyces pombe), or Trichoderma; an insect cell such as fall armyworm; a plant cell such as tobacco; or a mammalian cell such as BHK cells, CHO cells, COS cells, or myeloma cells. In some embodiments, the host cell is preferably a mammalian cell, more preferably a BHK cell, CHO cell, NSO cell, or COS cell.

[0110] According to the eleventh aspect, the present invention provides an antibody-drug conjugate (also known as an antibody-drug conjugate) comprising an anti-β-Klotho antibody or its antigen-binding moiety described in any of the first to seventh embodiments, which is bound to a therapeutic agent.

[0111] According to the twelfth aspect, the present invention provides a pharmaceutical composition comprising an anti-βKlotho antibody or its antigen-binding moiety according to any of the first to seventh aspects, or an antibody-drug conjugate according to the eleventh aspect, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is used to treat diseases mediated by βKlotho or βKlotho-FGFR1c-FGF21 (e.g., diseases requiring the mimicry or enhancement of the in vivo effects of FGF19 and / or FGF21). In some embodiments, the disease is a metabolic disorder and includes, but is not limited to, type 1 diabetes, type 2 diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

[0112] In some embodiments, the pharmaceutical composition may further contain one or more lubricants (e.g., talc, magnesium stearate, or mineral oil), wetting agents, emulsifiers, suspending agents, benzoic acid, preservatives (e.g., sorbic acid or calcium propionate), sweeteners, and / or flavoring agents. In some embodiments, the pharmaceutical composition according to the present invention may be prepared in the form of tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, suppositories, or capsules.

[0113] In some embodiments, the pharmaceutical compositions according to the present invention can be delivered using any physiologically acceptable method of administration, including but not limited to oral administration, extra-gastrointestinal administration, nasal administration, rectal administration, intraperitoneal administration, intravascular injection, subcutaneous administration, transdermal administration, and inhalation administration. In some embodiments, pharmaceutical compositions for therapeutic purposes can be prepared and stored in the form of lyophilized formulations or aqueous solutions by mixing reagents of a desired purity with optionally pharmaceutically acceptable carriers, excipients, etc.

[0114] According to the 13th aspect, the present invention provides the use in the manufacture of a pharmaceutical product for treating a disease mediated by βKlotho or βKlotho-FGFR1c-FGF21 (for example, a disease requiring the mimicry or enhancement of the in vivo effects of FGF19 and / or FGF21), an anti-βKlotho antibody or its antigen-binding moiety according to any of the 1st to 7th aspects, a nucleic acid molecule according to the 8th aspect, a vector according to the 9th aspect, a host cell according to the 10th aspect, or an antibody-drug conjugate according to the 11th aspect. In some embodiments, the disease is a metabolic disorder, including but not limited to type 1 diabetes, type 2 diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

[0115] According to the 14th aspect, the present invention provides a method for treating a disease in an individual that is mediated by βKlotho or βKlotho-FGFR1c-FGF21 (for example, a disease that requires the mimicking or enhancement of the in vivo effects of FGF19 and / or FGF21), comprising administering to the individual a therapeutically effective amount of an anti-βKlotho antibody or its antigen-binding moiety according to the 1st to 7th aspects, an antibody-drug conjugate according to the 11th aspect, or a pharmaceutical composition according to the 12th aspect. In some embodiments, the disease is a metabolic disorder and includes, but is not limited to, type 1 diabetes mellitus, type 2 diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, obesity, or any disease, disorder, or condition that requires the mimicking or enhancement of the in vivo effects of FGF19 and / or FGF21. In some embodiments of methods for treating or preventing a disease, disorder, or condition, the method comprises exposing cells to an anti-βKlotho antibody in vitro.

[0116] In some embodiments of the 12th to 14th aspects, the anti-βKlotho antibody or its antigen-binding moiety according to the present invention can be used alone or in combination with other compositions in a therapeutic setting. For example, the anti-βKlotho antibody can be administered with at least one additional therapeutic agent and / or adjuvant. In some embodiments, the additional compound is a therapeutic antibody other than the anti-βKlotho antibody, which includes, but is not limited to, agents for the treatment of diabetes, dyslipidemia, NASH, NAFLD, cardiovascular disease, metabolic syndrome, or obesity, such as biguanides, sulfonylureas, thiazolidinediones, GLP-1 analogs, PPARγ agonists, dipeptidyl peptidase-4 (DPP-4) inhibitors, bromocriptine preparations, bile acid chelating agents, insulin, α-glucosidase inhibitors, SGLT-2 inhibitors, appetite suppressants, and weight-loss agents, anti-inflammatory agents (e.g., aspirin, ibuprofen), etc.

[0117] According to the 15th aspect, the present invention provides a conjugate comprising an anti-βKlotho antibody or its antigen-binding moiety as described in any of the 1st to 7th aspects, and a detectable marker. The detectable marker is, for example, a detectable fluorescent marker, a chemiluminescent marker, or an isotope marker.

[0118] According to the sixteenth aspect, the present invention provides a fusion protein comprising an anti-βKlotho antibody or its antigen-binding moiety as described in any of the first to seventh aspects. Examples of antibody fusion proteins include Fab fusion proteins, Fc fusion proteins, and single-chain antibody (scFv) fusion proteins, which are named according to the site to which an effector protein (e.g., a cellular factor) is fused.

[0119] In some embodiments of the 15th or 16th aspect, a conjugate can be formed by synthesizing an anti-β-Klotho antibody according to the present invention covalently bound to a linker and one or more non-antibody agonists. In some embodiments, the antibody according to the present invention is conjugated or recombinantly fused with a diagnostic agent, detection agent or therapeutic agent, or any other molecule. The conjugated or recombinantly fused antibody can be used, for example, as part of a clinical trial method to monitor or predict the onset, occurrence, progression and / or severity of β-Klotho-mediated diseases, or to determine the effectiveness of a particular treatment. Such diagnosis and detection can be achieved, for example, by conjugating the antibody to a detectable marker. Detectable markers include, but are not limited to, enzymes, prosthetic groups, fluorescent markers, chemiluminescent markers, isotopic markers, and the like. In some embodiments, the antibody according to the present invention can be conjugated or recombinantly fused with a therapeutic or drug moiety that modifies a particular biological response (here, also the antibody-drug conjugate described in the 11th aspect). The therapeutic or drug moiety is not limited to typical chemotherapeutic agents. For example, the drug portion may be a protein, peptide, polypeptide, or low-molecular-weight toxin having the desired biological activity.

[0120] The above detailed description is intended only to further clarify the present invention to those skilled in the art, and should be understood not to limit any aspect. Those skilled in the art can make various modifications and changes to the above embodiments. [Examples]

[0121] The above detailed description is merely intended to make the content of the present invention clearer to those skilled in the art and is not intended to limit it to any particular embodiment. Those skilled in the art can make various changes and modifications to the embodiments. [Examples]

[0122] Antibody discovery based on phage display To obtain an antibody that specifically binds to human β-Klotho, the inventors performed phage display based on a human-derived natural single-chain antibody phage library.

[0123] First, human β-Klotho extracellular domain protein (R&D systems, 5889-KB-050) was biotinylated and used for liquid-phase panning. Subsequently, ProX-specific antibodies were screened using a human-derived natural single-chain antibody phage display bank (Shanghai Ruizhi Chemical Research Co., Ltd.). For each round of panning, the library was first incubated with Dynabeads streptavidin magnetic beads to remove nonspecific conjugates from the Dynabeads magnetic beads. Then, this library was panned with biotinylated human β-Klotho extracellular domain protein to enrich it with human β-Klotho-specific phage antibodies. Nonspecific phage antibodies were removed by multiple washes, and phage antibodies that specifically bind to human β-Klotho were eluted and recovered with trypsin or a known β-Klotho antibody (Tab hIgG1 antibody: its sequence is derived from 5H23 described in NGM patent CN201580016315.8). The eluted phages infected E. coli TG1, and with the help of helper phage M13KO7 (New Engineering BioLabs, Inc.), a library for the next round was generated. Typically, each round of panning includes negative selection, positive selection, washing, elution, and amplification. After each round of panning, based on the sequencing results, if most of the output clones are non-overlapping single clones, panning continues for up to 3 rounds. Otherwise, if the proportion of repetitive sequences is high, panning ends at this round. The phages output from each round are retained for subsequent use.

[0124] Monoclonals were selected from the panned library and grown overnight in 2YT medium containing 100 μg / mL ampicillin and 2% glucose. The overnight-cultured monoclonals were transferred to 2YT medium containing 100 μg / mL ampicillin. Inducible expression of scFv was performed by culturing in 1 mM IPTG at 30°C for 16 hours. The bacterial suspension was centrifuged and the supernatant was collected, and positive clones were screened using the following method.

[0125] Binding ELISA method: 100 μL of 1 μg / mL streptavidin was added to a 96-well plate and coated overnight at 4°C. After blocking at room temperature, 50 μL of 0.5 μg / mL biotinylated human β-Klotho extracellular domain protein was added and incubated at room temperature. After washing, 60 μL of blocking solution was added and incubated at room temperature. Then, 40 μL of scFv supernatant was added and incubated at room temperature. Tab hIgG1 antibody and hIgG1 isotype control at a final concentration of 1 μg / mL were added to the control wells, respectively. After washing, anti-Myc-HRP and anti-hIgG-Fc-specific-HRP were added and incubated at room temperature. After washing, 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB) substrate solution was added and incubated in the dark at room temperature for 15 minutes. The reaction was stopped by adding 100 μL of 1 M HCl, and the absorbance at 450 nm was read.

[0126] Competitive ELISA method: 100 μL of 1 μg / mL streptavidin was added to a 96-well plate and coated overnight at 4°C. After blocking at room temperature, 50 μL of 0.5 μg / mL biotinylated human βKlotho extracellular domain protein was added and incubated at room temperature. After washing, 60 μL of 3 μg / mL Tab hIgG1 was added and incubated at room temperature (the Tab hIgG1 and hIgG1 isotype antibody control wells were blocking solution). Then, 40 μL of scFv supernatant was added, and to the control wells, Tab hIgG1 and hIgG1 isotype antibody at a final concentration of 1 μg / mL each were added and incubated at room temperature. After washing, anti-Myc-HRP and anti-hIgG-Fc-specific-HRP were added and incubated at room temperature. After washing, 100 μL of TMB was added and incubated in the dark at room temperature for 15 minutes. 100 μL of 1 M HCl was added to stop the reaction, and the absorbance at 450 nm was read.

[0127] In the analysis of the results, OD during binding ELISA 450 > 0.4, and OD 450 (binding ELISA) / OD 450 (competitive ELISA) ratio ≥ 2, if it meets this condition, it can specifically bind to human βKlotho, and the antibody clone is considered to have an antigen-binding epitope similar to or identical to the Tab hIgG1 antibody. Also, OD during binding ELISA 450 > 0.4, and OD 450 (binding ELISA) / OD 450If the condition of a (competitive ELISA) ratio < 2 is met, it is considered an antibody clone that binds to an epitope different from that of the Tab hIgG1 antibody. Antibody clones similar to or identical to the Tab hIgG1 antibody-binding epitope, and clones with different partial epitopes were selected and sequenced to obtain 22 variable region sequences of human β-Klotho-binding antibodies, which were named CB-1, CB-2, CB-4, CB-5, CB-6, CB-8, CB-9, CB-10, CB-11, CB-14, CB-16, CB-17, CB-18, CB-19, CB-21, CB-22, CB-23, CB-24, CB-25, CB-26, CB-27, and CB-28, respectively. Then, full-length antibodies containing the above antibody variable regions were prepared using the human heavy chain IgG1 constant region and the human light chain λ or κ constant region. [Examples]

[0128] Screening of antibodies that activate human or monkey FGFR1c / βKlotho receptors using reporter assays. Based on a reporter gene assay, we measured whether the full-length antibody described in Example 1 could activate the human FGFR1c / βKlotho receptor. This method utilizes the principle that in a stable transgenic cell line, the human FGFR1c / βKlotho receptor is activated by FGF19 or the antibody, which then promotes the transmission of downstream signaling pathways, leading to ERK phosphorylation, further to Gal4-Elk1 phosphorylation, binding to the USA promoter region on DNA, and initiating transcription of the luciferase reporter gene. The activating effect of FGF19 or the antibody on the FGFR1c / βKlotho receptor was measured by measuring luciferase levels.

[0129] Stable transformed cell lines using reporter genes were constructed using the following method. First, two reporter plasmids, Gal4-Elk1 and 5xUAS-Luc, were introduced into rat L6 cells. After screening with antibiotics, cell lines stably expressing the target molecule were obtained. Next, plasmids encoding human FGFR1c and βKlotho genes or crab-eating monkey FGFR1c and βKlotho genes were further transfected, and after several antibiotic pressurized screenings, stable transformed FGFR1c / βKlotho / Gal4-Elk1 / Luc cell lines were obtained.

[0130] The function of the antibody in activating the human FGFR1c / βKlotho receptor was measured using the following method: 40 μL of a solution containing 3x the final concentration of antibody or FGF19 was added to a 96-well plate, followed by 80 μL of the above-mentioned stable transformed cells at an appropriate density, which were mixed with the antibody. The mixture was incubated overnight in an incubator at 37°C, 5% CO2 air, and saturated humidity. The following day, luciferase levels in the cells were detected using a One-Glo (Promega) kit. The following table shows data measuring the activation of the antibody's function of the human or monkey FGFR1c / βKlotho receptor.

[0131] [Table 1]

[0132] As shown in the results in Table 1, some of the anti-human βKlotho antibodies obtained by the phage display technology screening according to the present invention have an activating effect on humans and / or crab-eating monkeys βKlotho / FGFR1c. [Examples]

[0133] Detection of antibody binding to human β-Klotho, human α-Klotho, and mouse β-Klotho by ELISA. In this example, the binding of a portion of the antibody from Example 1 to human β-Klotho, human α-Klotho, and mouse β-Klotho was detected using the ELISA method. During detection, specific concentrations of extracellular domain protein solutions of human β-Klotho (R&D Systems), human α-Klotho (R&D Systems), or mouse β-Klotho (R&D Systems) were added to nickel-coated 96-well plates (Pierce) and incubated overnight at room temperature. The following day, after blocking the plates with PBS / 1% BSA, different concentrations of antibody were added to the reaction wells and incubated for 1.5 hours at room temperature. Then, HRP-labeled anti-human anti-human IgG Fab secondary antibody was added to the reaction wells and incubated for a further 30 minutes at room temperature. After adding the substrate and reacting at room temperature for 15 minutes, stop solution was added to the reaction wells. 450 The values ​​were read. The following are illustrative data regarding the binding of antibodies to human β-Klotho, human α-Klotho, and mouse β-Klotho in the ELISA assay.

[0134] [Table 2]

[0135] As shown in the data in Table 2, the anti-human βKlotho antibodies CB-1, CB-6, CB-8, CB-10, CB-16, CB-23, and CB-28 according to the present invention can all specifically bind to human βKlotho, while CB-1, CB-6, CB-8, and CB-10 do not bind to human αKlotho and exhibit high selective specificity for human-derived βKlotho protein. Furthermore, the partial antibody according to the present invention (e.g., CB-10) can also bind to mouse βKlotho. [Examples]

[0136] Affinity measurement of the antibody Biacore Based on the Biacore affinity assay, the binding affinity of four antibodies, CB-1, CB-6, CB-8, and CB-10, was measured. Binding affinity to human β-Klotho was evaluated by measuring the equilibrium dissociation constant (KD) of the purified antibodies. Specifically, anti-His tagged antibodies were immobilized on a CM5 tip flow cell using a His capture agent. Human β-Klotho extracellular domain protein (~80RU) was captured on the flow cell. Then, the purified antibody (in HEPES buffer containing 0.005% Tween20) was injected at a flow rate of 30 L / min, and binding dynamics were evaluated at 25°C. The results are as follows.

[0137] [Table 3]

[0138] As shown in the data in Table 3, the anti-human β-Klotho antibodies according to the present invention, such as CB-1, CB-6, CB-8, and CB-10, have a strong binding affinity for human β-Klotho. [Examples]

[0139] Modification of antibody affinity maturation The affinity maturation modification of the anti-human β-Klotho antibody according to the present invention was performed using the phage display method. The method of affinity maturation is described below using CB-8 as an example. The gene sequence of the CB-8 antibody variable region was constructed in a phage plasmid vector, and this was used as a template to construct the following four saturated mutant libraries. 1) A saturated mutant library of single-point mutation combinations, each containing one random mutant amino acid in either the light chain CDR1, light chain CDR3, or heavy chain CDR3. 2) A saturated mutant library of single-point mutation combinations containing one random mutant amino acid in either the light chain CDR2, heavy chain CDR1, or heavy chain CDR2. 3) A saturated mutant library of two-point mutation combinations of random mutations in two adjacent amino acids within the light chain CDR3. 4) A saturated mutant library of two-point mutation combinations, where two adjacent amino acids are randomly mutated within the heavy chain CDR3. Plasmids from these four saturated mutant libraries were constructed as phage libraries that met the library volume requirements, and the diversity of the phage libraries was determined by sequencing.

[0140] Phages from the above library were panned using human β-Klotho extracellular domain protein (Kactus Biosystems Co., Ltd.). Human β-Klotho extracellular domain protein was coated onto immunotubes, which were then blocked with 5% milk PBS blocking solution. A fixed concentration of phages was added to the immunotubes and incubated at room temperature for 2 hours. The immunotubes were washed multiple times with PBST, followed by three more washes with PBS. Finally, the phages bound to the human β-Klotho extracellular domain protein were eluted with 0.25% trypsin and recovered. The recovered phages were infected with E. coli SS320, and the phages were amplified. The amplified phages were recovered, and panning was performed again as described above. A total of three panning operations were performed.

[0141] Monoclonal phages were selected and amplified from the panned library, and positive clones that bound to human β-Klotho protein were screened using ELISA. The sequences of the positive clones were then determined. The method for measuring phage binding to human β-Klotho protein using ELISA was as follows: Human β-Klotho protein was coated onto a 96-well plate and then blocked with 5% milk PBS blocking solution. After adding the phage supernatant, the plate was incubated at room temperature for 1 hour, washed with PBST, and then HRP-labeled anti-M13 antibody was added and incubated for 1 hour. After multiple washes, TMB substrate solution was added and incubated at room temperature for 5-10 minutes. Finally, 2M H2SO4 was added to stop the reaction, and the absorbance at 450 nm was read. The titers of positive clones were measured, and then ELISA was used to detect the binding of phages with different titers to the human β-Klotho protein. The affinity of the positive antibodies was ranked using fitted binding curves. Based on the ranking results, the top eight antibody variable region sequences with the highest antibody binding positivity were selected (the sequences are shown in the table below).

[0142] Using the constant region of human heavy chain IgG1 and the constant region of human light chain κ, eight CB-8 antibody-derived antibody variable region sequences and the original CB-8 sequence were prepared as full-length antibodies. The sequences of the CDR region and variable region of the relevant sequence according to the present invention are as follows. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4]

[0143] The above antibodies were subjected to affinity testing using Gator (Gator bio). Briefly, the antibodies were first captured using a Protein A-coated probe (Gator bio), and the baseline was monitored for 60 seconds. Then, the binding of the antibodies to the antigen was measured at different concentrations (400 nM to 50 nM) of the human β-Klotho extracellular domain protein, and dissociation was detected in K-buffer (Gator bio). Simultaneously, the function of antibodies activating the human or crab-eating monkey FGFR1c / β-Klotho receptor was detected using the reporter gene assay described in Example 2. The results are shown in Table 5.

[0144] [Table 5]

[0145] As shown in Table 5, affinity maturation modification resulted in significantly improved binding affinity of all modified antibodies to human βKlotho, human βKlotho / FGFR1c, and crab-eating monkey βKlotho / FGFR1c compared to the unmodified antibody CB-8. [Examples]

[0146] Modification of latent PTM sites This invention involves detailed analysis of the CDR sequence of antibodies after affinity maturation, and the method for latent PTM site modification is described below using CB-8-24 as an example.

[0147] CDR sequence analysis revealed that CB-8-24 has a potential deamidation site NG in its heavy chain CDR2, but the corresponding site in CB-8-40 is SG, which can avoid the problem of potential deamidation. For example, a new antibody sequence CB-8-42 was designed considering the sequences of CB-8-24 and CB-8-40. Full-length antibodies including VH and VL of CB-8-42 were prepared using the human IgG1 (E233A / L235A) heavy chain constant region sequence and the human κ constant region sequence. [Table 6]

[0148] The constant region array is as follows: Heavy chain constant region sequence (HC): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO.50

[0149] Light chain constant region sequence (LC): RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO.51

[0150] The reporter gene assay described in Example 2 detected the function of the CB-8-42 full-length antibody in activating human or monkey FGFR1c / βKlotho receptors (see Table 6). The activation effect of CB-8-42 on FGFR1c / βKlotho receptors was significantly stronger than that of the control antibody, and in particular, the maximum cellular response to CB-8-42 was higher than that of the control antibody (results shown in Figure 1). Simultaneously, using the same method, the function of antibodies that activate FGFR2c / βKlotho, FGFR3c / βKlotho, and FGFR4 / βKlotho was detected. The results showed that CB-8-42 was unable to activate FGFR2c / βKlotho, FGFR3c / βKlotho, and FGFR4 / βKlotho receptors, demonstrating excellent specificity for human FGFR1c / βKlotho, as shown in Table 6.

[0151] [Table 7]

[0152] In summary, the anti-human β-Klotho antibody according to the present invention (particularly CB-8-42) is superior to the control antibody (5H23, whose sequence is derived from antibody 5H23 described in NGM patent CN201580016315.8) in terms of binding ability, binding specificity, or receptor activation ability. For example, as shown in Figure 1 and Table 6, the ability of CB-8-42 to activate human FGFR1c / β-Klotho is almost twice that of the control antibody, and the maximum relative activity is significantly superior to that of the control antibody (Figure 1). [Examples]

[0153] Animal in vivo drug efficacy research In this example, the in vivo efficacy of anti-human β-Klotho antibody was evaluated in an animal (crab-eating monkey) in vivo drug efficacy evaluation model. This invention investigates the potential metabolic-improving effect of anti-human β-Klotho antibodies (such as CB-8-42) in an obese crab-eating monkey model. The in vivo efficacy of the antibody according to this invention was investigated by monitoring changes in biochemical indicators such as food intake, body weight, BMI, fasting blood glucose, insulin, triglycerides, cholesterol, low-density lipoprotein, high-density lipoprotein, and liver fat content in crab-eating monkeys after antibody injection.

[0154] In the actual study, ten animals aged 11 years or older and weighing 8.5 kg or more were selected, and obese male crab-eating monkeys with MRI liver fat content ≥8% were identified. Based on body weight and liver fat content, they were randomly assigned to a solvent group (n=5) and an antibody group (n=5). A two-week adaptation period was then conducted, during which clinical observations and feeding statistics were performed daily, body weight measurements and BMI calculations were performed once a week, and basic blood biochemical indicators such as fasting blood glucose, insulin, triglycerides, cholesterol, low-density lipoprotein, and high-density lipoprotein were detected before administration. On days 1, 29, and 57 after the end of the two-week adaptation period, the animals were administered either the solvent (20 mM histidine buffer, 4.5% sorbitol, pH 5.7) or the anti-βKlotho antibody (CB-8-42, 10 mg / kg) by subcutaneous injection. During the study period, clinical observations and feeding statistics were performed daily, and body weight was measured once a week. Three weeks (day 22), seven weeks (day 50), and eleven weeks (day 78) after the initial administration, blood was collected from the animals after an overnight fast, and serum levels of blood glucose, insulin, triglycerides, cholesterol, low-density lipoprotein, and high-density lipoprotein were measured. Six weeks and twelve weeks after the initial administration, the liver fat content of the animals was measured using MRI.

[0155] Weight experiment results As shown in Figures 2-4, no significant changes were observed in the body weight of the animals in the solvent-treated group, but the body weight of the animals in the anti-human β-Klotho antibody CB-8-42 administration group gradually decreased, and by week 11 after administration, the body weight of the animals in the CB-8-42 administration group decreased by an average of 20%. Correspondingly, no significant changes were observed in the BMI of the animals in the solvent-treated group, but the BMI of the animals in the anti-human β-Klotho antibody CB-8-42 administration group decreased by an average of 20%.

[0156] Experimental results on food intake As shown in Figure 5, the food intake of the animals in the solvent-treated group remained basically stable throughout the experiment, at approximately 140 g / day. Animals administered with the anti-human β-Klotho antibody CB-8-42 showed a significant decrease in intake after administration. During the adaptation period, the average intake of the animals was approximately 112 g / day, and during the antibody treatment period, the average intake decreased to an average of 78 g / day, a decrease of approximately 30%.

[0157] Experimental results on liver fat content As shown in Figure 6, the liver fat content of animals in the solvent-treated group gradually increased, and there was a statistically significant difference in liver fat content after 12 weeks of treatment compared to before administration (p<0.05). After administration with the anti-human β-Klotho antibody CB-8-42, the mean liver fat content of the animals decreased from 12.1% before administration to 7.5% at 6 weeks after the first administration, and was maintained at 8.1% after 12 weeks. The liver fat content of animals in the antibody group before administration was equivalent to that of the solvent group, both at 12.1%. Twelve weeks after administration with the anti-human β-Klotho antibody CB-8-42, the liver fat content of animals in the antibody group according to the present invention was significantly lower than that of the solvent group (8.1% vs. 21.6%, p<0.01).

[0158] Experimental results of fasting blood glucose content As shown in Figure 7, the fasting blood glucose levels of the animals in the solvent-treated group gradually increased during the experiment, but after administration with the anti-human β-Klotho antibody CB-8-42, the animals' fasting blood glucose levels were significantly lower compared to before administration.

[0159] Experimental results of serum insulin levels As shown in Figure 8, serum insulin levels in the solvent-treated animals significantly increased during the experimental period, but after administration with the anti-human β-Klotho antibody CB-8-42, serum insulin levels in the animals significantly decreased.

[0160] Experimental results of serum triglyceride levels As shown in Figure 9, serum triglyceride levels in the solvent-treated animals increased significantly during the experiment, but after administration with the anti-human β-Klotho antibody CB-8-42, the animals' serum triglyceride levels remained stable.

[0161] Experimental results of serum total cholesterol levels As shown in Figure 10, serum total cholesterol levels in the solvent-treated animals remained stable throughout the experiment. However, after administration with the anti-human β-Klotho antibody CB-8-42, serum total cholesterol levels in the animals were significantly lower compared to before administration.

[0162] Experimental results of serum low-density lipoprotein levels As shown in Figure 11, serum low-density lipoprotein (LDL) levels in the solvent-treated animals remained substantially stable throughout the experiment. However, after administration with the anti-human β-Klotho antibody CB-8-42, the LDL levels in the animals were significantly reduced compared to before administration.

[0163] Experimental results of serum high-density lipoprotein levels As shown in Figure 12, serum high-density lipoprotein levels remained stable throughout the experiment in animals in the solvent-treated group and the anti-human β-Klotho antibody CB-8-42 administration group.

[0164] As can be seen from the combined results of the above experiments, using the anti-βKlotho antibody according to the present invention significantly improved the metabolic levels of crab-eating monkeys compared to the solvent group. Changes in various biochemical indicators such as feeding, body weight, BMI, fasting blood glucose, insulin, triglycerides, cholesterol, low-density lipoprotein, high-density lipoprotein, and liver fat content in crab-eating monkeys demonstrate the great potential and potential clinical therapeutic effect of the anti-human βKlotho antibody according to the present invention in the treatment or alleviation of diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity. Overall, the results of the drug efficacy experiments in animals showed that the anti-human βKlotho antibody according to the present invention significantly improved the above diseases in an obese crab-eating monkey model and demonstrated good in vivo safety.

[0165] Any embodiments or exemplary language provided herein (e.g., the use of "for example, etc." or "such as") are intended to better illustrate the invention and, unless otherwise claimed, do not limit the scope of the invention. The language herein should not be construed as indicating that any non-claimed element is necessary for the implementation of the invention.

[0166] All publications and patent applications cited herein are incorporated herein by reference and by means of such reference as each individual publication or patent application is specifically and individually indicated. Furthermore, any theories, mechanisms, demonstrations, or discoveries described herein are intended to further enhance the understanding of the present invention and are not intended to limit the invention in any way to such theories, mechanisms, demonstrations, or discoveries. The present invention has been shown and described in detail in the accompanying drawings and the foregoing, but should be considered illustrative and not limiting to the invention.

Claims

1. An anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, The heavy chain variable region includes heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 1, the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 38, the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 31, the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 4, the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 42, and the sequence of LCDR3 is the sequence indicated by SEQ ID NO.

6. Here, the sequences of HCDR and LCDR are defined by Kabat. or The sequence of HCDR1 is the sequence indicated by SEQ ID NO. 7, the sequence of HCDR2 is the sequence indicated by SEQ ID NO. 40, the sequence of HCDR3 is the sequence indicated by SEQ ID NO. 32, the sequence of LCDR1 is the sequence indicated by SEQ ID NO. 10, the sequence of LCDR2 is the sequence indicated by SEQ ID NO. 11, and the sequence of LCDR3 is the sequence indicated by SEQ ID NO.

6. Here, the sequences of HCDR and LCDR are defined by IMGT. Anti-βKlotho antibody or its antigen-binding portion.

2. The heavy chain variable region has an amino acid sequence represented by SEQ ID NO. 49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the amino acid sequence represented by SEQ ID NO. 49, and / or The light chain variable region has an amino acid sequence represented by SEQ ID NO. 44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the amino acid sequence represented by SEQ ID NO.

44. The anti-βKlotho antibody or its antigen-binding moiety according to claim 1.

3. The heavy chain variable region sequence is the sequence indicated by SEQ ID NO. 49, and the light chain variable region sequence is the sequence indicated by SEQ ID NO.

44. The anti-βKlotho antibody or its antigen-binding moiety according to claim 1.

4. An anti-βKlotho antibody or its antigen-binding moiety comprising a heavy chain variable region and a light chain variable region, The heavy chain variable region has an amino acid sequence represented by SEQ ID NO. 49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the amino acid sequence represented by SEQ ID NO.

49. The light chain variable region has an amino acid sequence represented by SEQ ID NO. 44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with the amino acid sequence represented by SEQ ID NO.

44. The heavy chain variable region includes heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), and the light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), The HCDR1, HCDR2, and HCDR3 are those included in the amino acid sequence shown in SEQ ID NO. 49, and the LCDR1, LCDR2, and LCDR3 are those included in the amino acid sequence shown in SEQ ID NO.

44. Anti-βKlotho antibody or its antigen-binding portion.

5. The anti-βKlotho antibody or its antigen-binding moiety binds to human βKlotho, or The anti-βKlotho antibody is a complete antibody, a single-chain antibody (scFv), a bispecific antibody, or a multispecific antibody, or The antigen-binding portion of the anti-βKlotho antibody is Fab, Fab', Fv, or F(ab'). ２ And, or The anti-βKlotho antibody is a mouse antibody, a chimeric antibody, or a humanized antibody, or The aforementioned anti-βKlotho antibody is a monoclonal antibody, or The anti-βKlotho antibody is an IgG1, IgG2, or IgG4 isotype, or The anti-βKlotho antibody is an IgG1 / IgG4, IgG2 / IgG4, or IgG1 / IgG2 chimeric type, or The anti-βKlotho antibody comprises a κ subtype or λ subtype light chain constant region, or The anti-βKlotho antibody comprises a human IgG1 heavy chain constant region and a human κ light chain constant region, or The anti-βKlotho antibody includes a wild-type Fc region or a genetically modified Fc region that has attenuated ADCC and / or CDC effects. The antibody or its antigen-binding portion according to claim 1.

6. The antibody has a heavy chain variable region indicated by SEQ ID NO. 49, a light chain variable region indicated by SEQ ID NO. 44, a heavy chain constant region indicated by SEQ ID NO. 50, and a light chain constant region indicated by SEQ ID NO.

51. The antibody or its antigen-binding portion according to claim 1.

7. An isolated nucleic acid molecule encoding the anti-βKlotho antibody or its antigen-binding moiety according to any one of claims 1 to 6.

8. A vector containing the nucleic acid molecule described in claim 7.

9. A host cell containing the nucleic acid molecule described in claim 7.

10. An antibody-drug conjugate comprising an anti-β-Klotho antibody or its antigen-binding moiety according to any one of claims 1 to 6, conjugated to a therapeutic agent.

11. A pharmaceutical composition comprising an anti-βKlotho antibody or its antigen-binding moiety according to any one of claims 1 to 6, and a pharmaceutically acceptable carrier.

12. The pharmaceutical composition according to claim 11, wherein the pharmaceutical composition is for treating diseases mediated by βKlotho or βKlotho-FGFR1c-FGF21.

13. The pharmaceutical composition according to claim 12, wherein the disease is a metabolic disorder.

14. The pharmaceutical composition according to claim 13, wherein the disease is type 1 diabetes, type 2 diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

15. The pharmaceutical composition according to claim 11 for use in the treatment of diseases mediated by βKlotho or βKlotho-FGFR1c-FGF21.

16. The pharmaceutical composition according to claim 15, wherein the disease is a metabolic disorder.

17. The pharmaceutical composition according to claim 16, wherein the disease is type 1 diabetes, type 2 diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, metabolic syndrome, or obesity.

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