FGF5 transcription variant-2 and its protein for the prevention, suppression, and therapeutic use of hepatic fibrosis.

Abstract

The present invention relates to the use of FGF5-v2 (FGF5 transcript variant 2) gene and its protein for preventing or treating liver fibrosis. The FGF5-v2 fragment gene (transcript and protein expressed therefrom) of the present invention is a mutant gene in which the amino acid sequence of fibroblast growth factor 5 (FGF5) is altered, and has the effect of suppressing the progression and proliferation of liver fibrosis. Therefore, FGF5-v2 is expected to prevent and treat liver fibrosis and block the progression to liver cirrhosis and liver cancer when administered to patients with fatty liver via a vector (e.g., AVV), etc., and is useful as a composition for treating liver fibrosis.

Description

Technical Field

[0001] The present invention relates to uses for the prevention, suppression, and treatment of liver fibrosis, such as transcript variants of FGF5 (fibroblast growth factor 5) and proteins expressed therefrom.

[0002] The present invention claims priority based on Korean Patent Application No. 10-2022-0159875 filed on November 25, 2022 and Korean Patent Application No. 10-2023-0156822 filed on November 13, 2023, and all the contents disclosed in the specifications and drawings of the applications are incorporated herein by reference.

Background Art

[0003] Liver fibrosis occurs from the excessive accumulation of extracellular matrix (including collagen) that occurs in most types of chronic liver diseases. Representative cells involved in liver fibrosis include hepatic stellate cells, Kupffer cells, endothelial cells, etc. Hepatic stellate cells are the main source of producing extracellular matrix and are involved in the increased production of various extracellular matrices including collagen. Kupffer cells are present in the sinusoidal space of the liver and substances produced by activated Kupffer cells come to affect surrounding hepatocytes, endothelial cells, and hepatic stellate cells, promoting liver fibrosis. Endothelial cells play an important role in regulating blood flow in the liver and are also involved in the production of growth factors and extracellular matrix related to the proliferation of hepatic stellate cells due to inflammation and liver fibrosis, etc. When liver fibrosis occurs, activated hepatic stellate cells increase geometrically, and the cells secrete profibrogenic cytokines, and as a result, produce extracellular matrix-related molecules such as α-SMA, collagen, and tissue inhibitor of metalloproteinase (TIMP).

[0004] In particular, among liver diseases, non-alcoholic fatty liver disease (NAFLD) is a disease that progresses to non-alcoholic steatohepatitis (NASH) and cirrhosis, and is highly associated with metabolic diseases such as obesity, diabetes, and hyperlipidemia, which are not caused by alcohol consumption. Non-alcoholic steatohepatitis is recognized as a serious disease worldwide because fat accumulates in liver cells while degeneration / necrosis of liver cells occurs, leading to inflammation and liver fibrosis, which develops into cirrhosis (or hepatic sclerosis) and liver cancer. NASH develops from two separate events that progress from steatosis to fibrosis, and this is called the "two-hit process" (Non-Patent Literature 1).

[0005] On the other hand, fibroblast growth factor (FGF) is a type of growth factor, forming a family of 23 types. However, it is known that each type has diverse in vivo functions and activities, and their specific roles have not been fully elucidated. In this context, FGF5 (fibroblast growth factor 5) has been known to be involved in the progression of hepatic fibrosis in non-alcoholic steatohepatitis (NASH) (Non-Patent Literature 1). However, the present inventors have confirmed that a newly identified FGF5 mutant can effectively suppress fibrosis in the liver. Focusing on this, they have confirmed the potential of the FGF5 mutant as a liver disease treatment agent based on its excellent anti-fibrotic effect. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] U.S. Patent Application No. 08 / 991,601 [Patent Document 2] U.S. Patent Application No. 08 / 409,297 [Patent Document 3] U.S. Patent No. 5,608,149 [Patent Document 4] U.S. Patent No. 5,608,144 [Patent Document 5] U.S. Patent No. 5,604,121 [Patent Document 6] U.S. Patent No. 5,569,597 [Patent Document 7] U.S. Patent No. 5,466,785 [Patent Document 8] U.S. Patent No. 5,399,680 [Patent Document 9] U.S. Patent No. 5,268,463 [Patent Document 10] U.S. Patent No. 5,608,142 [Patent Document 11] European Patent No. 307,247 [Patent Document 12] International Publication No. 91 / 08291 [Non-patent literature]

[0007] [Non-Patent Document 1] Exp Anim. 2014;63(1):85-92. [Non-Patent Document 2] Dayhoff et al., in Atlas of Protein Sequence and Structure, vol 5, supp 3, 1978 [Overview of the project] [Problems that the invention aims to solve]

[0008] In addition, while researching therapeutic strategies that can control liver fibrosis, the inventors of the present invention discovered that, contrary to the existing knowledge that FGF5 (fibroblast growth factor 5) is involved in the progression of liver fibrosis in non-alcoholic steatohepatitis (NASH), FGF-v2 (FGF5 transcript variant 2), which is an FGF5 transcript variant and a fragment (short variant) of FGF5 according to the present invention, significantly suppresses liver fibrosis, thus completing the present invention.

[0009] Therefore, the object of the present invention is to provide an FGF5-v2 transcript variant and its protein (also referred to as "FGF5-v2 protein") consisting of the amino acid sequence of Sequence ID No. 1 in the sequence catalog.

[0010] Another object of the present invention is to provide a nucleic acid molecule (i.e., the gene for FGF5-v2) that encodes the FGF5-v2 protein. Another object of the present invention is to provide a transduction medium comprising the FGF5-v2 gene, its transcript, or its protein (or a transduction medium on which the gene, transcript, or protein is supported); or a composition comprising the transduction medium.

[0011] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of hepatic fibrosis (or hepatic fibrosis), comprising as an active ingredient an FGF5-v2 protein consisting of the amino acid sequence of Sequence ID No. 1 in the sequence catalog of the present invention; an expression vector containing a nucleic acid molecule encoding the FGF5-v2 protein; or isolated cells into which the expression vector has been introduced.

[0012] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those with ordinary skill in the art to which the present invention pertains from the following description. [Means for solving the problem]

[0013] In order to achieve the above object, the present invention provides an FGF5-v2 protein consisting of the amino acid sequence of SEQ ID NO: 1 in the sequence list.

[0014] Furthermore, the present invention provides a nucleic acid molecule encoding the above FGF5-v2 protein. In one embodiment of the present invention, the FGF5-v2 gene may include, but is not limited to, the nucleotide sequence of SEQ ID NO: 2 in the sequence list.

[0015] Furthermore, the present invention provides an expression vector containing the above nucleic acid molecule. Furthermore, the present invention provides a cell into which the above expression vector has been introduced. Preferably, the cell is an isolated cell.

[0016] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating liver fibrosis, which contains, as an active ingredient, an FGF5-v2 protein consisting of the amino acid sequence of SEQ ID NO: 1 in the sequence list; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or an isolated cell into which the expression vector has been introduced.

[0017] In one embodiment of the present invention, the FGF5-v2 protein, the nucleic acid molecule, the expression vector, or the cell may be carried by a carrier, but is not limited thereto.

[0018] In another embodiment of the present invention, the composition contains, as an active ingredient, an FGF5-v2 protein, the nucleic acid molecule, or the expression vector, and the carrier may be one or more selected from the group consisting of virus particles, vesicles, nanoparticles, microparticles, liposomes, transposons, micelles, antibodies, and exosomes, but is not limited thereto.

[0019] In yet another embodiment of the present invention, the hepatic fibrosis may be, but is not limited to, hepatic fibrosis in one or more conditions selected from the group consisting of alcoholic fatty liver disease, non-alcoholic fatty liver disease, hepatitis C, hepatitis B, autoimmune hepatitis, hepatic encephalopathy, primary biliary sclerosis, liver cancer, and hepatocellular carcinoma.

[0020] In yet another embodiment of the present invention, the non-alcoholic fatty liver disease may be one or more selected from the group consisting of non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), gepatic steatosis, acute fatty liver of pregnancy (AFLP), NAFLD-related liver failure, NAFLD-related liver fibrosis, NAFLD-related cirrhosis, and NAFLD-related liver cancer, but is not limited to these.

[0021] In other embodiments of the present invention, the composition may be administered in combination with an anti-inflammatory agent, but is not limited thereto. In yet another embodiment of the present invention, the anti-inflammatory agent may be a steroidal anti-inflammatory agent or a nonsteroidal anti-inflammatory agent, but is not limited to these.

[0022] Furthermore, the present invention provides a method for preventing or treating hepatic fibrosis, comprising the steps of administering to an individual in need an FGF5-v2 protein consisting of the amino acid sequence of Sequence ID No. 1 in a sequence catalog; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced. However, the FGF5-v2 gene (nucleic acid molecule), transcript, and / or protein of the present invention may be administered to an individual using a variety of carriers (transmitters) known in the art other than a vector. Examples of such carriers include expression vectors, viruses, vesicles, nanoparticles, microparticles, liposomes, transposons, micelles, exosomes, antibodies, and the like.

[0023] Furthermore, the present invention provides for the use of a composition comprising, as an active ingredient, an FGF5-v2 protein consisting of the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced, for the prevention or treatment of hepatic fibrosis.

[0024] Furthermore, the present invention provides for the use of a composition comprising, as an active ingredient, an FGF5-v2 protein consisting of the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced, for the production of a prophylactic or therapeutic preparation for liver fibrosis.

[0025] Furthermore, the present invention provides uses for the prevention or treatment of hepatic fibrosis of an FGF5-v2 protein comprising the amino acid sequence of Sequence ID No. 1 in the sequence catalog; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced.

[0026] Furthermore, the present invention provides uses for the production of agents for the prevention or treatment of hepatic fibrosis, including an FGF5-v2 protein comprising the amino acid sequence of Sequence ID No. 1 in a sequence catalog; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced. [Effects of the Invention]

[0027] The fibroblast growth factor 5 (FGF5) transcription mutant FGF5-v2 and its protein according to the present invention were developed by modifying the amino acid sequence of full-length FGF5. It has been confirmed that the transcription mutant and its protein suppress the progression of liver fibrosis and improve liver function abnormalities associated with liver fibrosis, as well as increases in various inflammation and fibrosis-related cytokines. Therefore, it is expected that administering the gene encoding FGF5-v2, its transcription, protein, and / or various carriers (e.g., AAV) carrying these to patients can suppress liver fibrosis and achieve preventive, ameliorative, and therapeutic effects against a variety of diseases, including cirrhosis and liver cancer. [Brief explanation of the drawing]

[0028] [Figure 1] This invention presents a recombinant expression vector for intracellular transmission and expression (transcript or protein expression) of the FGF5-v2 variant (referred to as "FGF5-v2" or "FGF5-S") encoding gene. [Figure 2a] This schematic diagram shows a series of experimental schedules for creating an in vivo model of liver fibrosis (an in vivo NAFLD mouse model induced by a CDAA-HFD diet) and confirming the liver fibrosis-inhibiting effect of the FGF5-v2 mutant of the present invention (referred to as "FGF5-v2" or "FGF5-S"). [Figure 2b]This shows the changes in body weight over time after injecting a transduction medium containing the full-length FGF5 gene ("FGF5-F") or the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") into a NAFLD mouse model (Normal: normal mice; Control: untreated control group; the same applies below). [Figure 2c] In normal mice, a control group, an FGF5-F administered group, and an FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") administered group, changes in glucose tolerance were observed after 12 hours of fasting followed by glucose injection. [Figure 3a] This is a comparison of NAFLD activity scores (NAS) after injecting FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S") into NAFLD mouse models. [Figure 3b] This study compared fibrosis scores after injecting FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S") into NAFLD mouse models, specifically confirming the inhibitory effect of injecting the FGF5-v2 mutant ("FGF5-v2" or "FGF5-S") on liver fibrosis. [Figure 4a] This study involved injecting FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S") into NAFLD mouse models, followed by a comparison of AST levels in the serum of NAFLD mouse models. The results specifically confirmed the AST suppression effect (i.e., liver damage and liver function improvement effect) achieved by injecting the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"). [Figure 4b]This study involved injecting FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S") into NAFLD mouse models, followed by a comparison of ALT levels in the serum of NAFLD mouse models during liver function assessment. The results specifically confirmed the ALT suppression effect (i.e., liver damage and liver function improvement effect) of injecting the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"). [Figure 4c] This is the result of examining the serum AST / ALT ratio of NAFLD mouse models after injecting them with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") gene. [Figure 4d] This study compares the levels of triglycerides (TG) in the serum of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). [Figure 4e] This graph shows a comparison of HDL (High-density lipoprotein) levels in the serum of NAFLD mouse models after injecting them with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") gene. [Figure 4f] This graph shows a comparison of LDL (Low-density lipoprotein) levels in the serum of NAFLD mouse models after injecting them with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") gene. [Figure 4g] This graph shows a comparison of glucose levels in the serum of NAFLD mouse models after injecting them with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") gene. [Figure 5a]This graph shows a comparison of α-SMA levels in liver tissue mRNA of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). The inhibitory effect of injecting the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") specifically on α-SMA was confirmed at the mRNA level. [Figure 5b] This graph shows a comparison of Col1α1 levels in fibrosis-related biomarkers in liver tissue mRNA of NAFLD mouse models after injecting the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") gene into the NAFLD mouse model. The specific inhibitory effect of injecting the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on Col1α1 was confirmed. [Figure 5c] This graph shows a comparison of fibronectin levels in liver tissue mRNA of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). [Figure 5d] This graph shows a comparison of Vimentin levels in fibrosis-related biomarkers in liver tissue mRNA of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). [Figure 5e] The following shows the results of measuring the expression levels of FGF5-F (Figure 5e) and FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") transcripts in NAFLD mice (n=8 per group) injected with the FGF5-F gene or the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") (Figure 5f). [Figure 5f] Same as above. [Figure 6a]This paper presents the results of quantitative analysis of the expression of the inflammatory marker IL-1β protein using ELISA experiments with liver tissue from NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). [Figure 6b] This paper presents the results of quantitative analysis of IL-6 protein expression, an inflammatory marker, using ELISA experiments with liver tissue from NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). [Figure 6c] The following results show the analysis of α-SMA protein expression levels using Western blot in liver tissue of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). The specific inhibitory effect of injecting the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on α-SMA protein levels was confirmed. [Figure 7a] This study shows the results of comparing the expression of various cytokines and chemokines in the serum of NAFLD mouse models after injecting them with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"). From these results, the inhibitory effect of the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on liver fibrosis was confirmed. [Figure 7b] This study compares the protein expression levels of various cytokines and chemokines in the liver tissue of NAFLD mouse models after injecting them with FGF5-F or FGF5-v2 mutants (referred to as "FGF5-v2" or "FGF5-S"). The results confirm the suppressive effect of the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on liver fibrosis-related cytokines. [Figure 8a]This image shows the results of Sirius red staining of liver tissue sections from a NAFLD mouse model injected with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"), confirming the inhibitory effect of the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on liver fibrosis. [Figure 8b] This image shows the results of H&E staining of liver tissue sections from a NAFLD mouse model injected with the FGF5-F or FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"). [Figure 9] This figure shows the full-length FGF5 sequence, with the regions corresponding to the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") (underlined) and FGF5-v3 (boxed). [Figure 10a] This graph shows the results of stimulation experiments using human hepatic stellate cell line (LX2 cells). The x-axis represents the time (minutes) after FGF5 treatment, and the y-axis represents the fold value relative to the control experiment. The full-length FGF5 protein was confirmed to have signaling activity through the Erk phosphorylation reaction. [Figure 10b] This graph shows the results of stimulation experiments using human hepatic stellate cell lines (LX2 cells). The x-axis represents the time after protein treatment with the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S"), and the y-axis represents the fold value relative to the control experiment. Through the Erk phosphorylation reaction, it was confirmed that the FGF5-v2 mutant protein (referred to as "FGF5-v2" or "FGF5-S") does not exhibit signal transduction activity. [Figure 10c]This graph shows the results of stimulation experiments using human hepatic stellate cell lines (LX2 cells). The x-axis represents the results of pretreatment with 1, 3, and 5 times the concentration of FGF5-v2 mutant protein (referred to as "FGF5-v2" or "FGF5-S") relative to FGF5 treatment, and the y-axis represents the fold value relative to the control experiment. It was confirmed that the FGF5-v2 mutant protein (referred to as "FGF5-v2" or "FGF5-S") antagonizes the signaling activity of full-length FGF5 protein. From this, the inhibitory effect of the FGF5-v2 mutant (referred to as "FGF5-v2" or "FGF5-S") on liver fibrosis was confirmed. [Modes for carrying out the invention]

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as would be commonly understood by those skilled in the art. In this invention, the term "protein" is used interchangeably with "polypeptide" and refers to a polymer of amino acid residues, as is commonly found in naturally occurring proteins.

[0030] This invention provides a mutant gene for FGF5 (fibroblast growth factor 5), an FGF5 transcript variant expressed therefrom, and a protein translated therefrom. Specifically, the present invention aims to provide the FGF5-v2 (FGF5 transcript variant 2) gene, a newly discovered FGF5 fragment, the transcript expressed therefrom, and a protein translated therefrom, etc.

[0031] The FGF5-v2 variant of the present invention is a newly discovered FGF5 fragment. It can be referred to as "FGF5-v2" or "FGF5-S". The inventors have newly discovered that the FGF5-v2 protein not only differs in its properties from the full-length FGF5 protein and other fragments (FGF5-v1 or FGF5-v3), but also has completely different effects from the parent protein FGF5, including improvement of liver fibrosis and therapeutic effects. The full-length FGF5 is a known protein, and specific information can be found in public databases such as NCBI (registration number NM_004464.4).

[0032] The FGF5-v2 protein of the present invention is characterized by containing or consisting of the amino acid sequence of Sequence ID No. 1 in the sequence catalog. In the present invention, FGF5-v2 protein means including its functional equivalent. The functional equivalent is a polypeptide having at least 50%, 70%, preferably 80%, and more preferably 90% sequence homology (i.e., identity) with the amino acid sequence constituting the FGF5-v2 protein (preferably, as a given example, the amino acid sequence shown as Sequence ID No. 1 in the sequence catalog). For example, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, The present invention refers to a polypeptide having sequence homology of 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100%, and exhibiting substantially the same physiological activity as the ancestral FGF5-v2 protein (preferably, as a prime example, the polypeptide shown as SEQ ID NO: 1 in the sequence catalog). Here, "substantially the same physiological activity" preferably means a preventive, ameliorative, or therapeutic effect on liver fibrosis. Preferably, in the present invention, the functional equivalent of the FGF5-v2 protein may be produced as a result of the addition, substitution, or deletion of a portion of the amino acid sequence of SEQ ID NO: 1 in the sequence catalog. In the above, the amino acid substitution is preferably a conservative substitution. Examples of naturally occurring conservation substitutions of amino acids include: aliphatic amino acids (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn), and sulfur-containing amino acids (Cys, Met). Furthermore, functional equivalents of the FGF5-v2 protein also include variants in which some amino acids are deleted from the amino acid sequence of the FGF5-v2 protein (i.e., even shorter functional fragments).The deletion or substitution of the aforementioned amino acids is preferably located in a region not directly related to the physiological activity of the FGF5-v2 protein. That is, the deletion or substitution is carried out within a range that does not affect the physiological activity of the FGF5-v2 protein. In addition, variants in which several amino acids are added to both ends or within the sequence of the FGF5-v2 protein amino acid sequence are also included. Furthermore, the scope of functional equivalents of the present invention also includes polypeptide derivatives in which the chemical structure of a part of the polypeptide is modified while maintaining the basic backbone of the FGF5-v2 protein and its physiological activity. For example, this includes structural modifications to change the stability, storability, volatility, or solubility of the protein.

[0033] In this specification, sequence homology and homogeneity are defined as the percentage of identical matching residues (amino acid residues or bases) in the candidate sequence relative to the original sequence, after aligning the candidate sequence with the original sequence (preferably, SEQ ID NO: 1 in the sequence catalog for amino acid sequences, or, for nucleic acid sequences, preferably, SEQ ID NO: 2 in the sequence catalog) and introducing gaps. Where necessary, conservative substitutions are not considered as part of the sequence homogeneity to obtain the maximum percentage sequence homogeneity. Furthermore, in determining the homology or homogeneity of protein sequences, the N-terminus, C-terminus, or internal extensions, deletions, or insertions of the FGF5-v2 protein amino acid sequence are not interpreted as sequences that affect sequence homogeneity or homology. Moreover, such sequence homogeneity can be determined by common standard methods used to compare similar portions of the amino acid sequences of two polypeptides. Computer programs such as BLAST or FASTA align the two polypeptides so that their respective amino acids are optimally matched (along the full-length sequences of one or two sequences, or along predicted portions of one or two sequences). The program provides a default opening penalty and a default gap penalty, and offers a scoring matrix such as PAM250 (Standard Scoring Matrix; Dayhoff et al., in Atlas of Protein Sequence and Structure, vol 5, supp 3, 1978) that can be used in conjunction with the computer program. For example, percentage homogeneity can be calculated as follows: Multiply the total number of identical matches by 100, and then divide by the length of the longer sequence within the corresponding span and the sum of the number of gaps introduced in the longer sequence to align the two sequences.

[0034] Furthermore, the present invention provides an FGF5-v2 gene encoding the FGF5-v2 protein of the present invention. In this invention, the terms “nucleic acid,” “gene sequence,” “DNA sequence,” “RNA sequence,” or “polynucleotide” refer to deoxyribonucleotides or ribonucleotides in single or double-stranded form. Unless otherwise specified, this also includes known analogs of naturally occurring nucleotides that are hybridized to nucleic acids in a manner similar to naturally occurring nucleotides.

[0035] In one embodiment of the present invention, the FGF5-v2 gene may be characterized by containing the nucleotide sequence of Sequence ID No. 2 in the sequence catalog. In another embodiment of the present invention, the FGF5-v2 gene may consist of the nucleotide sequence of Sequence ID No. 2 in the sequence catalog.

[0036] In the present invention, the polynucleotide encoding the target protein may undergo various modifications to the coding region, either due to codon degeneracy or considering the preferred codon in the organism in which the protein is to be expressed, as long as these modifications do not alter the amino acid sequence of the protein expressed from the coding region. Similarly, the portion excluding the coding region may undergo various modifications as long as these do not affect gene expression, and those skilled in the art will understand that such modified genes are also included within the scope of the present invention. That is, the polynucleotide of the present invention may be mutated by substitution, deletion, insertion, or combination thereof of one or more nucleic acid bases, as long as it encodes a protein having equivalent activity, and these are also included within the scope of the present invention.

[0037] In other words, the nucleic acid molecule (gene) indicated by a specific sequence in this specification may include not only that sequence but also its biological equivalents. That is, considering variants with biologically equivalent activity of nucleic acid molecules, a uniform nucleic acid molecule is interpreted to include sequences that exhibit substantial identity with the sequence number listed in the sequence catalog. Specifically, a nucleic acid molecule containing a nucleotide sequence indicated by a specific sequence number is not limited to that nucleotide sequence alone, and variants of the nucleotide sequence are included within the scope of this invention. In the sequence catalog of this invention, a nucleic acid molecule consisting of a nucleotide sequence indicated by a specific sequence number is a concept that includes functional equivalents of the nucleic acid molecule that constitutes it, for example, variants in which a part of the nucleotide sequence of the nucleic acid molecule has been altered by deletion, substitution, or insertion, but which are functionally identical to the nucleic acid molecule. Specifically, the nucleic acid molecules disclosed in the present invention may include nucleotide sequences having sequence homology of 70% or more, more preferably 80% or more, even more preferably 90% or more, and most preferably 95% or more, with the amino acid sequences indicated by specific sequence numbers in the sequence catalog. For example, this includes nucleic acid molecules having sequence homology of 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100%. The "percentage of sequence homology" for nucleic acid molecules is determined by comparing two optimally sequenced sequences with a comparison region, where some of the nucleotide sequences in the comparison region may contain additions or deletions (i.e., gaps) compared to the reference sequence (which does not contain additions or deletions) of the optimal sequences of the two sequences.

[0038] The FGF5-v2 mutant gene (nucleic acid molecule), its transcript, and / or its protein of the present invention can be transmitted to an individual via a variety of carriers (transmitters) or directly by themselves. The carrier is sufficient as long as it transmits the FGF5-v2 gene, transcript, and / or protein to a host cell or other target organism, causing the gene to be expressed in the cell or the transcript or protein to exert its intrinsic physiological activity (liver fibrosis inhibitory activity), and is not limited to any specific type. For example, expression vectors, viruses (viral particles), vesicles, nanoparticles, microparticles, liposomes, transposons, micelles, antibodies, or exosomes can be used as carriers for transmitting the FGF5-v2 mutant, but are not limited thereto, and gene, transcript, and protein transmission methods known in the art can be applied without limitation. Furthermore, the protein can be transmitted directly together with a preservative.

[0039] For example, the present invention provides an expression vector (recombinant expression vector) containing the FGF5-v2 gene (i.e., a nucleic acid molecule encoding the FGF5-v2 protein) according to the present invention. That is, the present invention provides an expression vector (recombinant expression vector) containing a promoter; and the FGF5-v2 gene (FGF5-v2 polynucleotide) operably linked thereto.

[0040] In this invention, the term "expression" means the production of proteins or nucleic acids in cells. In the present invention, "recombinant vector" refers to a vector capable of expressing a peptide or protein encoded by a different nucleic acid inserted within the vector, and preferably means a vector manufactured to express a target protein (in the present invention, FGF5-v2 protein). The "vector" refers to any medium for the introduction and / or transfer of bases to a host cell in vitro, in vitro, or in vivo, and may be a replication unit to which other DNA fragments bind, resulting in the replication of the bound fragment. "Replication unit" refers to any genetic unit (e.g., plasmid, phage, cosmid, chromosome, virus, etc.) that functions as a self-unit of DNA replication in vivo, that is, a unit capable of replicating under its own regulation.

[0041] The vector according to the present invention may be, but is not limited to, linear DNA, plasmid DNA, or recombinant viral vector. The vector includes, for example, plasmid vectors, cosmid vectors and bacteriophage vectors, adenovirus vectors, lentivirus vectors, retrovirus vectors, and viral vectors such as adenovirus-related viral vectors. Preferably, the vector is an adenovirus vector. The inventors used the pADTrack vector in specific examples.

[0042] The term "promoter" refers to a DNA sequence that regulates the expression of a nucleic acid sequence operably linked in a specific host cell. It may further include an optional operator sequence for regulating transcription, a sequence encoding an appropriate mRNA-ribosome binding site, and a sequence regulating the termination of transcription and sequencing. The promoter can be a constitutive promoter that continuously induces the expression of a target gene at all times, or an inducible promoter that induces the expression of a target gene at a specific location or time. Examples include the SV40 promoter, CMV promoter, CAG promoter, CaMV 35S promoter, Rsyn7 promoter (US Patent Application No. 08 / 991,601), rice actin promoter, ubiquitin promoter, and ALS promoter (US Patent Application No. 08 / 409,297). In addition, all promoters disclosed in U.S. Patents 5,608,149; 5,608,144, 5,604,121, 5,569,597, 5,466,785, 5,399,680, 5,268,463 and 5,608,142, etc., may be used.

[0043] The recombinant vector of the present invention may include, in addition to the promoter, any operator sequence for regulating transcription, a sequence encoding a suitable mRNA-ribosome binding site and a sequence regulating the termination of transcription and decoding, a terminator, and more preferably a polyhistidine tag (an amino acid motif composed of at least five histidine residues), a signal peptide gene, an endoplasmic reticulum retention signal peptide, a cloning site, and may further include a tag gene, a selection marker gene such as an antibiotic resistance gene for selecting transformants, and so on. In the recombinant vector, the polynucleotide sequence of each gene is operatively linked to the promoter. As used herein, "operatively linked" means a functional link between a nucleotide expression regulatory sequence, such as a promoter sequence, and a different nucleotide sequence, so that the regulatory sequence regulates the transcription and / or decoding of the other nucleotide sequence.

[0044] The recombinant vector can be constructed using prokaryotic or eukaryotic cells as a host. For example, if the vector of the present invention is an expression vector and uses prokaryotic cells as a host, it generally includes a strong promoter to advance transcription (e.g., pLλ promoter, trp promoter, lac promoter, tac promoter, T7 promoter, etc.), a ribosome binding site for decryption disclosure, and a transcription / decryption termination sequence. If eukaryotic cells are used as a host, the replication origins on which the vector operates in eukaryotic cells may include, but are not limited to, the f1 replication origin, SV40 replication origin, pMB1 replication origin, adeno replication origin, AAV replication origin, and BBV replication origin. Furthermore, promoters derived from mammalian cell genomes (e.g., metallothione promoter) or mammalian viruses (e.g., late adenovirus promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus promoter, and HSV tk promoter) are used, and generally have a polyadenylated sequence as the transcription termination sequence.

[0045] Typical examples of the aforementioned tag genes include Avi tag, Calmodulin tag, polyglutamate tag, E tag, FLAG tag, HA tag, His tag (polyhistidine tag), Myc tag, S tag, SBP tag, IgG-Fc tag, CTB tag, Softag 1 tag, Softag 3 tag, Strep tag, TC tag, V5 tag, VSV tag, and Xpress tag. Preferably, the vector according to the present invention may include the myc tag.

[0046] On the other hand, the expression vector can be introduced into target cells (host cells) as an expression vector by methods known to the art, such as infection, transfection, or transduction.

[0047] The gene transfer method using plasmid expression vectors is an FDA-approved method for human use that directly transfers plasmid DNA to mammalian cells. Plasmid DNA has the advantage of being homogeneously purified, unlike viral vectors. The plasmid expression vectors that can be used in this invention are mammalian expression plasmids known in the industry. For example, but not limited to these, pRK5 (European Patent No. 307,247), pSV16B (International Patent Publication No. 91 / 08291), and pVL1392 (PharMingen) are typical examples. The plasmid expression vector can be introduced into target cells by methods known to the art, including but not limited to transient transfection, microinjection, transduction, cell fusion, calcium phosphate precipitation, liposome-mediated transfection, DEAE dextran-mediated transfection, polybrene-mediated transfection, electroporation, gene guns, and other known methods for introducing DNA into cells.

[0048] Furthermore, as methods applicable to the present invention, the viral expression vectors containing the nucleic acids include, but are not limited to, retroviruses, adenoviruses, herpesviruses, avipoxviruses, lentiviruses, and the like. The retroviral vectors are manufactured (fabricated) in which the viral genes are completely removed or modified so that non-viral proteins are produced in cells infected with the viral vector. The main advantages of retroviral vectors for gene therapy are that they can deliver a large amount of gene into replicating cells, accurately integrate the gene delivered into the cellular DNA, and do not cause continuous infection after gene transfection. The retroviral vectors certified by the FDA were manufactured using PA317 amphotropic retrovirus packaged cells. An example of a non-retroviral vector is the adenovirus mentioned above. The main advantage of adenoviruses is that they can carry a large amount of DNA fragment (36kb genome) and have the ability to infect non-replicating cells with very high titers. Furthermore, herpes viruses can also be usefully used for human gene therapy. In addition, other known and suitable viral vectors can be used in the present invention.

[0049] If the expression vector is a viral recombinant vector, the vector can be transfected or transfected into virus-producing cells, i.e., a packaging cell line. Several diverse techniques commonly used to introduce exogenous nucleic acids (DNA or RNA) into prokaryotic or eukaryotic host cells can be used for transfecting or transfecting. The virus containing the target gene (FGF5-v2 gene) according to the present invention is grown in the packaging cell line and released extracellularly, and the virus can be transductioned into the target cells. The nucleic acid of the virus transductioned into the cell is either inserted into the cell's genome or used to produce the target transcript or protein (FGF5-v2 transcript or protein) without being inserted.

[0050] Furthermore, isolated cells into which the expression vector according to the present invention has been introduced (transformed, transfected, or transfused) can be provided. The cells refer to cells for growing (amplifying) the expression vector. That is, the cells represent host cells into which the aforementioned nucleic acid molecule or expression vector has been directly transfected / transformed / transfected. For example, if the expression vector is a viral vector, the cells may be packaging cells for producing a virus containing the viral vector. The selection of an appropriate host should be obvious to the ordinarily skilled in the art from the implications of this application.

[0051] To achieve yet another objective of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of hepatic fibrosis (hepatic fibrosis), comprising, as an active ingredient, an expression vector containing an FGF5-v2 transcript, an FGF5-v2 protein, a nucleic acid molecule encoding the FGF5-v2 protein; and / or isolated cells into which the expression vector has been introduced. In addition to the components mentioned, the pharmaceutical composition may include a variety of carriers for delivering the FGF5-v2 gene, transcript, and / or protein of the present invention to target cells. That is, the pharmaceutical composition may include the FGF5-v2 gene, transcript, and / or protein supported on a carrier. A specific description of the carrier has been given above and is therefore omitted.

[0052] In this invention, "liver fibrosis (liver fibrotic disease)" refers to a disease in which the liver is impaired in form and function due to the accumulation of fibrosis caused by persistent damage to liver tissue from various causes. The term "liver fibrosis" is used interchangeably with "liver fibrosis." Fibrosis is a phenomenon in which excessive fibrous connective tissue is formed in organs and tissues during the wound healing process in response to repeated damage to cells and tissues. The accumulation of persistent liver fibrosis is a cause of various liver diseases, including liver cancer and cirrhosis. Therefore, suppressing and improving liver fibrosis can prevent the development of serious diseases and treat liver dysfunction. The inventors have confirmed that FGF5-v2 of this invention effectively suppresses fibrosis in liver tissue in animal models of liver disease and improves the disease by suppressing the expression of cytokines related to fibrosis and inflammation.

[0053] In one embodiment of the present invention, the liver fibrosis includes, but is not limited to, alcoholic fatty liver disease, non-alcoholic fatty liver disease, hepatitis C, hepatitis B, autoimmune hepatitis, hepatic encephalopathy, primary biliary sclerosis, liver cancer, and hepatocellular carcinoma, as well as all other liver fibrosis or chronic liver diseases of unknown cause or details. It may also include any disease directly or indirectly caused by or accompanied by liver fibrosis.

[0054] In one embodiment of the present invention, the non-alcoholic fatty liver disease may be one or more selected from the group consisting of non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), hepatic steatosis, acute fatty liver of pregnancy (AFLP), NAFLD-related liver failure, NAFLD-related liver fibrosis, NAFLD-related cirrhosis, and NAFLD-related liver cancer, but is not limited thereto.

[0055] In another embodiment of the present invention, the pharmaceutical composition of the present invention (i.e., FGF5-v2 protein or its expression vector, etc.) is characterized by being administered in combination with an anti-inflammatory agent, but is not limited thereto. That is, the FGF5-v2 of the present invention can achieve a superior anti-fibrotic effect when used in combination with existing anti-inflammatory agents.

[0056] As used herein, the term "combination therapy" refers to the administration of individual components of a therapy simultaneously, sequentially, or individually. Combination therapy is the process of obtaining a synergistic effect by administering two or more drugs simultaneously or sequentially, or by administering them alternately at fixed or unspecified intervals. Combination therapy can be defined, but is not limited to, a synergistic effect that provides therapeutic superiority over the efficacy obtained by administering one or the remaining components of the combination therapy at normal doses, as measured over the degree of response, response rate, time to disease progression, or survival period.

[0057] The pharmaceutical composition of the present invention can be administered simultaneously, separately, or sequentially with an anti-inflammatory agent. Even when administered sequentially with an anti-inflammatory agent, the order of administration is not restricted. However, the administration method may be appropriately adjusted depending on the type of anti-inflammatory agent, the patient's condition, the patient's sex, age, etc.

[0058] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of hepatic fibrosis, comprising (i) an expression vector containing the FGF5-v2 protein of the present invention, a nucleic acid molecule encoding it, or isolated cells into which the expression vector has been introduced; and (ii) an anti-inflammatory agent as an active ingredient.

[0059] The composition may be in the form of a mixture containing (i) FGF5-v2 protein and (ii) an anti-inflammatory agent, or it may be in a form for simultaneous administration of (i) FGF5-v2 protein and (ii) the anti-inflammatory agent. For example, (i) FGF5-v2 protein and (ii) the anti-inflammatory agent may be a single dosage form produced by mixing them with a pharmaceutically acceptable adjuvant, diluent, or carrier. In this case, (i) FGF5-v2 protein and (ii) the anti-inflammatory agent may each contain various components within the scope of the respective drug. Furthermore, the single dosage form may be a separate dosage form from such a dosage form and can be administered together with other preparations containing other therapeutic agents.

[0060] Alternatively, the composition may be in a form in which (i) FGF5-v2 protein and (ii) an anti-inflammatory agent are each formulated and administered simultaneously, separately, or sequentially. In this case, the composition may be a combination-administered pharmaceutical composition for simultaneous or sequential administration, comprising a first pharmaceutical composition containing a pharmaceutically effective amount of (i) FGF5-v2 protein as an active ingredient, and a second pharmaceutical composition containing a pharmaceutically effective amount of (ii) an anti-inflammatory agent as an active ingredient.

[0061] The anti-inflammatory agent according to the present invention may be a steroidal anti-inflammatory agent and / or a non-steroidal anti-inflammatory agent. The non-steroidal anti-inflammatory agent can be selected from salicylic acid, acetic acid, propionic acid, mefenamic acid, the oxicam series, or the non-acidic series. Specific examples include, but are not limited to, aspirin, ibuprofen, naproxen, ketoprofen, and fenoprofen.

[0062] Furthermore, the present invention provides a kit for the prevention or treatment of hepatic fibrosis, comprising a pharmaceutical composition according to the present invention. The kit according to the present invention is not limited in its specific form as long as it is for the prevention or treatment of hepatic fibrosis, and may without limitation include any components and equipment for the manufacture, storage, expression, administration, etc., of the FGF5-v2 gene, transcript, protein, and expression vector according to the present invention.

[0063] The content of the FGF5-v2 gene, its transcript, its protein, and / or expression vector in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the patient's condition, etc. For example, it may be 0.0001 to 99.9% by weight or 0.001 to 50% by weight based on the weight of the total composition, but is not limited thereto. The content ratio is a value based on the dry weight after removing the solvent.

[0064] The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions. The excipient may be one or more selected from the group consisting of, for example, diluents, binders, disintegrants, lubricants, adsorbents, humectants, film coating substances, and controlled-release additives.

[0065] The pharmaceutical compositions according to the present invention can be used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, elixirs, emulsions, suspensions, alcoholic preparations, lozenges, aromatic preparations, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, soft extracts, dried extracts, liquid extracts, injections, capsules, perfusion solutions, ointments, lotions, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols, respectively, according to conventional methods. The external preparations may have dosage forms such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasms.

[0066] Examples of carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0067] When formulating, the product is typically prepared using fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. The following are additives for tablets, powders, granules, capsules, pills, and lozenges according to the present invention: corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, monocalcium phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, excipients such as propylene glycol, casein, calcium lactate, Primogel; gelatin, gum arabic, ethanol, agar powder, cellulose phthalate acetate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch paste, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, and other binders can be used, including hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum (Guar Disintegrants such as gum, baking soda, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium aluminum silicate, D-sorbitol solution, and light anhydrous silicic acid;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ethers, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid can be used.

[0068] The following can be used as additives to the liquid formulation according to the present invention: water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamin, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, and the like.

[0069] The syrup according to the present invention may contain a solution of sucrose, other sugars, or sweeteners, and may also contain fragrances, colorants, preservatives, stabilizers, suspending agents, emulsifiers, viscosity modifiers, etc., as needed.

[0070] The emulsion according to the present invention can use purified water, and emulsifiers, preservatives, stabilizers, fragrances, etc., can be used as needed. The suspending agent according to the present invention can be acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropyl methylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, and other suspending agents, and surfactants, preservatives, stabilizers, colorants, and fragrances can be used as needed.

[0071] The injectable preparations according to the present invention include solvents such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG (PEG), Ringer's lactate injection, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidinedione, propylene glycol, twins, nicotinamide, hexamine, and dimethylacetamide; weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), and organic compounds. It may also contain buffering agents such as compounds, proteins, albumin, peptones, and gums; isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; sulfurizing agents such as 0.1% sodium bisulfite, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and sodium acetone bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium CMC, sodium alginate, Twin 80, and aluminum monostearate.

[0072] The suppositories according to the present invention include cocoa butter, lanolin, witepsol, polyethylene glycol, glycerol gelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, coconut oil, cocoa butter + cholesterol, lecithin, lanette wax, glycerol monostearate, twin or span, Imhausen, monolen (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrokote SP, S-70-XXA, S-70-XX75 (S-70-XX95), and Hydrokote. Bases such as 25, Hydrocote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Maspol, Maspol-15, Neospostal-en, Paramount-B, Sposeal (OSI, OSIX, A, B, C, D, H, L), Suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Spostal (N, Es), Wecoby (W, R, S, M, Fs), and Tegestor triglyceride base (TG-95, MA, 57) can be used.

[0073] Solid formulations for oral administration include tablets, pills, powders, granules, and capsules. Such solid formulations are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose, or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.

[0074] Liquid formulations for oral administration include suspensions, oral solutions, emulsions, and syrups. Besides commonly used simple diluents such as water and liquid paraffin, various excipients may be included, such as humectants, sweeteners, fragrances, and preservatives. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0075] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level can be determined by factors including the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and elimination ratio, the duration of treatment, drugs used concurrently, and other factors well known in the medical field.

[0076] The pharmaceutical compositions according to the present invention can be administered as individual therapeutic agents or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered single or multiple times. It is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, taking all of the above factors into consideration, and this can be easily determined by a person of ordinary skill in the art to which the present invention belongs.

[0077] The pharmaceutical composition of the present invention is administered to an individual via a variety of routes. All methods of administration are predictable, but for example, it can be administered by oral ingestion, subcutaneous injection, intraperitoneal injection, intravenous injection, intramuscular injection, perispinal space (intradural) injection, sublingual administration, buccal mucosal administration, rectal insertion, vaginal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, etc.

[0078] The pharmaceutical composition of the present invention is determined by the type of active ingredient drug, along with various related factors such as the disease being treated, the route of administration, the patient's age, sex, weight, and the severity of the disease. Specifically, the effective amount of the composition according to the present invention varies depending on the patient's age, sex, and weight, and is generally 0.001 to 150 mg per kg of body weight, preferably 0.01 to 100 mg, administered daily or every other day, or divided into 1 to 3 doses per day. However, the dosage can be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc., and therefore, the aforementioned dosage does not limit the scope of the present invention in any way.

[0079] In this invention, "individual" means an object that requires treatment for a disease, and more specifically, it means a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, and cow.

[0080] In the present invention, "administration" means providing a predetermined composition of the present invention to an individual by any appropriate method. In the present invention, "prevention" means any action that suppresses or delays the onset of the target disease; "treatment" means any action by which the administration of the pharmaceutical composition according to the present invention improves or beneficially alters the target disease and the metabolic abnormalities caused thereby; and "improvement" means any action by which the administration of the composition according to the present invention reduces parameters related to the target disease, such as the severity of symptoms.

[0081] The following examples of preferred embodiments and experimental cases are provided to aid in understanding the present invention. However, the following embodiments and experimental cases are provided only to facilitate understanding of the present invention, and the content of the present invention is not limited by the following embodiments and experimental cases.

[0082] Example 1. Preparation for the production of FGF5-v2 (FGF5 transcript variant 2) and gene injection. Using the restriction enzymes KpnI and HindII, the pAdTrack-CMV vector was used to synthesize genes corresponding to FGF5 variant 2, specifically PN 5'-AAAGGTACCATGAGCTTGTCCTTCCTCCTC-3'primer (sequence number 19 in the sequence catalog) and 5'-AAAAAGCTTTCACTTATCGTCGTCATCCTTGTAATCTCTGTGAACTTGGCTTAACATATTGGCTTCGT-'primer (sequence number 20 in the sequence catalog). After cleavage with the restriction enzymes KpnI and HindII, ligation was performed. The manufactured recombinant vector (pAdTrack-CMV hFGF5-2) was transformed into E. coli for amplification, and the pAdTrack-CMV hFGF5-2 vector and pAdEASY vector were cleaved with PmeI restriction enzyme. These were then used to transform E. coli BJ5183 cells, and transformed E. coli BJ5183 cells expressing hFGF5-2 were selected. Subsequently, after isolating the target gene, adenovirus hFGF5-2 was produced using 293T cells.

[0083] Example 2. Production of an in vivo diet-induced non-alcoholic fatty liver disease (NAFLD) model and gene injection as a liver fibrosis model. Male C57BL / 6J mice (6 weeks old) free of specific pathogens were purchased from DBL (Chungcheong Province, South Korea) and adapted to the rearing environment one week before the start of treatment. At 7 weeks of age, the mice were randomly divided into two groups. For 6 weeks, the normal group (n=8) was fed a standard commercial diet, and the liver fibrosis group (n=24) was fed CDAA-HFD (A06071302, Research Diets, NJ, USA). CDAA-HFD is a high-fat diet containing L-amino acids and methionine that is deficient in choline, and induces NAFLD (Nonalcoholic fatty liver disease; simple fatty liver, steatohepatitis, and liver fibrosis (cirrhosis associated with non-alcoholic fatty liver)).

[0084] The liver fibrosis group was further divided into an empty vector administration group; a full-length FGF5 (also called FGF5-F; FGF5 full sequence) administration group; and an FGF5 mutant FGF5-v2 (also called FGF5-S) administration group for further experiments. Therefore, the pAd-GFP-control group (n=8), pAd-hFGF5 (FGF5-F expression vector) (n=8), or pAd-hFGF5-v2 (FGF5-v2 expression vector) (n=8) prepared in Example 1 were divided into 1 × 10⁻⁶ groups. 9 The PBS was administered individually via the tail vein in a single injection. A group of normal mice not fed a high-fat diet were given PBS. All mice underwent glucose tolerance testing after 12 days, and were euthanized with CO2 two days later. All mice were weighed weekly. The mice were housed in a chamber with controlled temperature (23±3°C) and relative humidity (40-60%), free from specific pathogens.

[0085] Example 3. Confirmation of the effect of FGF5-v2 gene injection on suppressing liver fibrosis in an in vivo NAFLD mouse model. As described above, normal mice or NAFLD mice were treated with an empty vector (control), full-length FGF5 (FGF5-F), or FGF5-v2 (FGF5-S), and after undergoing a glucose tolerance test (GTT), liver tissue was collected from each mouse, and the NAS score and Fibrosis sicore, which are indicators of fatty liver, were measured. Specifically, the degree of steatosis and fibrosis in the tissue was evaluated by a pathologist who scored the stained mouse tissue by looking at photographs. Steatosis was numerically scored according to quantitative pathological standards (steatosis, inflammation, and hepatocyte swelling). Fibrosis was evaluated by staining mouse liver tissue fragments with Sirius Red, observing them, and scoring them into stages F0 (no fibrosis), F1 (mild), F2 (moderate), F3 (fibrotic crosslinking), and F4 (cirrhosis).

[0086] First, the results of the steatosis evaluation are shown in Figure 3a. NAFLD mice supplied with CDAA-HFD showed a significant increase in the NAS score, an indicator of fatty liver, compared to mice supplied with a normal diet. Among NAFLD mice, no significant difference in NAS score was observed between the untreated control group, the FGF5-F treated group, and the FGF5-S treated group. On the other hand, when comparing the degree of fibrosis, as can be seen in Figure 3b, the degree of fibrosis, which was significantly increased by the CDAA-HFD diet, did not differ significantly from the untreated control group when treated with full-length FGF5-F. However, when treated with FGF5-v2, the FGF5 transcription mutant of the present invention, the degree of fibrosis was greatly reduced. From these results, it was confirmed that FGF5-v2 has a particularly significant improvement effect on liver fibrosis that appears in fatty liver, rather than on fatty liver itself.

[0087] Example 4. Confirmation of suppression of non-alcoholic fatty liver fibrosis in serum of an in vivo NAFLD mouse model by FGF5-v2 gene injection. In this example, the effect of the FGF5 transcription variant of the present invention on suppressing liver fibrosis was confirmed through serum analysis of a mouse model. Specifically, the serum levels of biomarkers indicating the degree of fatty liver fibrosis were quantified.

[0088] Specifically, mice were anesthetized with isoflurane using a respiratory anesthesia machine, euthanized, and blood was collected by cardiac puncture. The collected blood was centrifuged at 4,000 rpm for 30 minutes at 4°C to collect serum, which was then stored at -80°C and used for subsequent analysis. The levels of all metabolism-related factors present in the serum (AST, ALT, TG, HDL, LDL, and glucose, etc.) were measured in the serum samples prepared as described above using the Advia 1800 Chemistry System according to the manufacturer's protocol (Siemens Healthcare, Erlangen, Germany).

[0089] As a result, no significant difference was observed in HDL and LDL between the untreated control group and the full-length FGF5 or FGF5-v2 treated groups (Figures 4e and 4f). However, analysis of AST and ALT, which are used as hepatocyte damage markers in fatty liver disease, showed that these levels were significantly reduced in the FGF5-v2 treated group compared to the untreated control group and the full-length FGF5 treated group (Figures 4a-4c). Furthermore, it was confirmed that triglycerides (TG) and glucose were also significantly reduced in the FGF5-v2 treated group compared to the other groups (Figures 4d and 4g). These results indicate that the FGF5-v2 of the present invention has a particularly excellent effect in suppressing liver fibrosis in diseases related to non-alcoholic fatty liver disease.

[0090] Example 5. Confirmation of suppression of liver fibrosis markers in liver tissue mRNA of an in vivo NAFLD mouse model by FGF5-v2 gene injection. In this example, mRNA levels of fibrosis-related factors were analyzed in the liver tissue of each mouse model, and the inhibitory effect of the FGF5 transcription mutant of the present invention on liver fibrosis was confirmed at the RNA level.

[0091] RNA was isolated using the RNeasy Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturer's protocol. RNA quality, total RNA yield, and 260 / 280 and 260 / 230 ratios were measured using a NanoDrop spectrophotometer (Thermo, Massachusetts, USA). Total RNA was isolated using PrimeScript according to the manufacturer's protocol (Takara, Shiga, Japan). TMThe first strand cDNA was reverse transcribed using a 1st strand cDNA Synthesis Kit with complementary DNA. Quantification of the complementary DNA template was performed by real-time PCR using SYBR green fluorescence (Takara) on a CFX384 instrument (Bio-Rad, California, USA). Primer information is as follows: mα-SMA forward 5'-TGC TGA CAG AGG CAC CAC TGA A-3' and reverse 5'-CAG TTG TAC GTC CAG AGG CAT AG-3'; mCol1α1 forward 5'-CCT CAG GGT ATT GCT GGA CAA C-3' and reverse 5'-CAG AAG GAC CTT GTT TGC CAG G-3'; mFibronectin forward 5'-CCC TAT CTC TGA TAC CGT TGT CC-3' and reverse 5'- TGC CGC AAC TAC TGT GAT TCG G-3'; mVimentin forward 5'-CGG AAA GTG GAA TCC TTG CAG G-3' and reverse 5' AGC AGT GAG GTC AGG CTT GGA A-3'; mHPRT forward 5' CTG GTG AAA AGG ACC TCT CGA AG-3' and reverse 5'-CCA GTT TCA CTA ATG ACA CAA ACG-3'.

[0092] As a result, α-SMA and Col1α1, the main markers of fibrosis, were significantly reduced in the FGF5-v2 treatment group compared to the control group and the FGF5(Full) treatment group (Figures 5a and 5b). In the case of Fibronectin and Vimentin, also known as EMT markers, the FGF5(Full) treatment group showed increased levels compared to the untreated control group, while the FGF5-v2 treatment group showed levels similar to the control group (Figures 5c and 5d). These results indicate that the FGF5 transcription variant of the present invention can suppress the expression of liver fibrosis-related factors and inhibit liver fibrosis in non-alcoholic fatty liver disease. On the other hand, when the expression of full-length FGF5 and FGF5-v2 transcripts was measured in each mouse model into which the full-length FGF5 or FGF5-v2 gene was introduced, it was confirmed that in the group injected with the full-length FGF5 gene, no FGF5-v2 transcripts were detected, and only full-length FGF5 was detected, while in the group injected with the FGF5-v2 gene, no full-length FGF5 transcripts were detected, and only FGF5-v2 was detected (Figures 5e and 5f).

[0093] Example 6. Confirmation of suppression of liver fibrosis at liver tissue protein levels in an in vivo NAFLD mouse model by FGF5-v2 gene injection. In this example, the protein levels of fibrosis-related factors were analyzed in the liver tissue of each mouse model using ELISA and Western blot, and the inhibitory effect of the FGF5 transcription mutant of the present invention on liver fibrosis was confirmed at the protein level.

[0094] Specifically, liver tissue isolated from mice was dissolved in T-PER buffer (Thermo, Massachusetts, USA) supplemented with PhosSTOP and a protease inhibitor (Roche, Basel, Switzerland), and centrifuged at 15,000 rpm for 10 minutes at 4°C. Protein concentration was quantified by BCA analysis (Thermo). The amounts of IL-6 and IL-1β (R&D Systems, Massachusetts, USA) were measured using ELISA techniques on tissue proteins of a certain concentration, or through immunoblotting experiments using SDS-PAGE. Antibodies such as α-SMA (abcam, Cambridge, UK) and β-actin (Cell signaling, Massachusetts, USA) were used.

[0095] As a result, while there was no significant difference in the FGF5-v2 treatment group compared to the control group for the inflammatory markers IL-1β and IL-6 (see Figures 6a and 6b), we were able to confirm that the fibrosis marker α-SMA was significantly reduced in the mouse model injected with FGF5-v2 compared to the other groups (Figure 6c). These results indicate that in non-alcoholic fatty liver disease in mouse models, administration of the FGF5 transcription mutant of the present invention is particularly effective in reducing liver fibrosis compared to improving inflammation.

[0096] Example 7. Confirmation of cytokine and chemokine protein expression in serum and liver tissue of an in vivo NAFLD mouse model with FGF5-v2 gene injection. In this example, cytokine and chemokine levels were analyzed in serum and liver tissue of each mouse model to confirm the therapeutic effect of administering the FGF5 transcription variant of the present invention.

[0097] Specifically, serum was obtained from each mouse as described in Example 4, and liver tissue fractions were obtained using the same method as described in Example 6. Cytokine and chemokine protein expression levels were measured using the MILLIPLEX MAP Mouse Cytokine / Chemokine Kit (R&D systems). Fluorescence intensity was interpreted using MAGPIX® (Luminex Corporation, Austin, Texas).

[0098] As a result, as can be seen in Figure 7a, NAFLD mice administered with FGF5-v2 showed significantly reduced levels of cytokines and chemokines (CCL2, CCL5, CCL11, and CXCL2) secreted by hepatic stellate cells, the main cells associated with hepatic fibrosis, compared to other NAFLD mouse groups. Furthermore, we were able to confirm that CXCL1 and fibroblast growth factor 21 (FGF21), which are involved in fibrosis, were also significantly reduced in the FGF5-v2 administered group. In addition, the FGF5-v2 administered group showed higher serum levels of the anti-inflammatory cytokine IL-10 compared to the other groups.

[0099] As can be seen in Figure 7b, the expression of CCL2, CCL5, CCL11, and CXCL2 in liver tissue was reduced in the FGF5-v2 administered group compared to the other groups, and the level of fibroblast growth factor 21 (FGF21) was also significantly reduced.

[0100] The results indicate that the FGF5 transcription variant of the present invention can suppress the production and secretion of cytokines and chemokines that promote fibrosis in non-alcoholic fatty liver disease, and increase the levels of anti-inflammatory cytokines, thereby improving and treating liver fibrosis.

[0101] Example 8. Comparison of liver tissue staining in an in vivo NAFLD mouse model with FGF5-v2 gene injection (Sirius red staining and H&E staining). In this example, the degree of progression of hepatic fibrosis was confirmed in liver tissue of a NAFLD mouse model through Sirius red staining, which is known as an indicator of fibrosis, and the degree of progression of fatty liver disease was confirmed through H&E staining.

[0102] Specifically, as in the previously described examples, liver tissue specimens were obtained from each mouse, fixed in 10% buffered formalin, embedded in paraffin, and then cut into 5 μm thick sections to obtain tissue sections. Each tissue section was stained with Sirius Red using the Piclosirius Red Staining Kit (abcam) as instructed by the manufacturer. H&E staining was performed according to standard procedures.

[0103] The results of Sirius red staining are shown in Figure 8a, and the results of H&E staining are shown in Figure 8b. Comparative analysis of liver tissue stained with Sirius red in tissue sections, magnified to 100x or 400x, showed no significant difference in the degree of Sirius red staining between the liver tissue of the group injected with the FGF5 full gene and the untreated control group. On the other hand, the liver tissue of mice injected with the FGF5-v2 gene showed a significantly reduced degree of Sirius red staining compared to the other groups (Figure 8a). These results indicate that FGF5-v2 can effectively suppress fibrosis in the liver.

[0104] Comparative histopathological analysis of liver tissue treated with H&E staining, magnified to 100x or 400x, confirmed that there was no significant difference in the degree of steatosis between the liver tissue of the group injected with the FGF5 full gene, the group injected with the FGF5-v2 gene, and the untreated control group (Figure 8b). These results suggest that the FGF5-v2 gene has a specific and significant improvement effect on liver fibrosis, which is a symptom of fatty liver, rather than having an effect on fatty liver itself.

[0105] Example 9. Confirmation of FGF5 antagonistic activity of FGF5-v2 protein through experiments on hepatic stellate cell lines. In this example, stimulation experiments were conducted using a hepatic stellate cell line (LX2 cells), and the inhibitory effect of the FGF5 transcription mutant of the present invention on liver fibrosis was confirmed at the protein level.

[0106] Hepatic stellate cells (LX2 cells) were similarly seeded the day before, then subjected to a starvation process for about 1 hour the following day, followed by treatment with recombinant protein FGF5 at 100 ng / ml. Total ERK and phospho-ERK protein levels were checked at the time point. Hepatoblastoma cells (HepG2) were also used as a control.

[0107] As a result, although the total ERK level did not change, we were able to confirm that phospho-ERK levels increased in the group treated for only 5 minutes, and gradually decreased in the group treated for 10 minutes or longer (Figure 10a). However, there was not much difference in hepatoblastoma (HepG2).

[0108] Furthermore, to investigate the effects of FGF5-v2, when only FGF5-v2 protein (FGF5s) was treated under the same conditions as described above, the hepatic stellate cell stimulation response was requantified using the phospho-ERK reaction, and no significant difference in the response level was observed compared to the mutant protein (Figure 10b).

[0109] Then, under similar conditions, FGF5 and FGF5-v2 protein (FGF5s) were treated together at 1x, 3x, and 5x concentrations. As a result, when FGF5 alone was treated, phospho-ERK levels increased, but in the group treated with both FGF5 and FGF5-v2, phospho-ERK levels decreased. Furthermore, it was confirmed that when the amount of FGF5-v2 was increased compared to when the same amount was treated, phospho-ERK levels decreased even more, thus confirming that FGF5-v2 has an antagonistic effect on FGF5 (Figure 10c). No changes were observed in hepatoblastoma (HepG2) treated in the same manner.

[0110] After correcting the experimental results of hepatic stellate cells (LX2 cells) using semi-quantitative values ​​with total ERK, a significant difference was observed when phospho-ERK expression levels were checked. This result is similar to the mechanism that could be inferred from the animal experimental results performed above, and indicates that the FGF5-v2 protein is effective in suppressing liver fibrosis by antagonizing the effect of FGF5 on hepatic stellate cells.

[0111] As can be seen from the above examples, the FGF5 transcription mutant, FGF5-v2, according to the present invention can effectively suppress and improve liver fibrosis by suppressing the levels of pro-fibrosis factors and increasing the levels of anti-inflammatory cytokines in a non-alcoholic fatty liver animal model. Therefore, it is expected that FGF-v2 of the present invention will be useful in the prevention and treatment of various liver fibrosis-related diseases such as cirrhosis and liver cancer through its excellent liver fibrosis inhibitory effect.

[0112] The above description of the present invention is illustrative, and a person with ordinary skill in the art to which the present invention belongs will understand that it can be easily modified in other specific forms without changing the technical idea or essential features of the present invention. Therefore, it should be understood that the embodiments described above are merely illustrative and not limiting.

[0113] The sequence information related to the present invention is shown in Table 1 below.

[0114] [Table 1-1]

[0115] [Table 1-2]

[0116] [Table 1-3]

[0117] [Table 1-4] [Industrial applicability]

[0118] The fibroblast growth factor 5 (FGF5) transcription mutant FGF5-v2 and its protein according to the present invention were developed by modifying the amino acid sequence of full-length FGF5. It has been confirmed that the transcription mutant and its protein suppress the progression of liver fibrosis and improve liver function abnormalities associated with liver fibrosis, as well as increases in various inflammation and fibrosis-related cytokines. Therefore, it is expected that administering the gene encoding FGF5-v2, its transcription, protein, and / or various carriers (e.g., AAV) carrying these to patients can suppress liver fibrosis and achieve preventive, ameliorative, and therapeutic effects against various diseases, including cirrhosis and liver cancer, thus suggesting potential for industrial application.

Claims

1. A pharmaceutical composition for the prevention or treatment of liver fibrosis, comprising as an active ingredient: an FGF5-v2 protein having the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced.

2. The pharmaceutical composition according to claim 1, characterized in that the FGF5-v2 protein, the nucleic acid molecule, the expression vector, or the cell is supported on a transduction vessel.

3. The pharmaceutical composition according to claim 2, wherein the composition comprises the FGF5-v2 protein, the nucleic acid molecule, or the expression vector as an active ingredient, and the transduction body is one or more selected from the group consisting of viral particles, vesicles, nanoparticles, microparticles, liposomes, transposons, micelles, antibodies, and exosomes.

4. The pharmaceutical composition according to claim 1, characterized in that the liver fibrosis is liver fibrosis in one or more conditions selected from the group consisting of alcoholic fatty liver disease, non-alcoholic fatty liver disease, hepatitis C, hepatitis B, autoimmune hepatitis, hepatic encephalopathy, primary biliary sclerosis, liver cancer, and hepatocellular carcinoma.

5. The pharmaceutical composition according to claim 4, characterized in that the non-alcoholic fatty liver disease is one or more selected from the group consisting of non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), hepatic steatosis, acute fatty liver of pregnancy (AFLP), NAFLD-related liver failure, NAFLD-related liver fibrosis, NAFLD-related cirrhosis, and NAFLD-related liver cancer.

6. The pharmaceutical composition according to claim 1, characterized in that the composition is administered in combination with an anti-inflammatory agent.

7. The pharmaceutical composition according to claim 6, characterized in that the anti-inflammatory agent is a steroidal anti-inflammatory agent or a non-steroidal anti-inflammatory agent.

8. A method for preventing or treating hepatic fibrosis, comprising the steps of: administering an FGF5-v2 protein consisting of the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced to a non-human individual requiring such treatment.

9. Use of a composition comprising, as an active ingredient, an FGF5-v2 protein having the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced, for the prevention or treatment of hepatic fibrosis in a non-human organism.

10. Use of a composition comprising, as an active ingredient, an FGF5-v2 protein having the amino acid sequence of SEQ ID NO: 1; a nucleic acid molecule encoding the FGF5-v2 protein; an expression vector containing the nucleic acid molecule; or isolated cells into which the expression vector has been introduced, for the production of a prophylactic or therapeutic preparation for hepatic fibrosis.

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