KR102517456B1 - Antiviral composition comprising fibroblast growth factor 11 as an active ingredient - Google Patents

Abstract

The present invention relates to an antiviral composition comprising fibroblast growth factor 11 as an active ingredient, wherein fibroblast growth factor 11 (FGF11) is used to treat hepatitis B virus (HBV ), inhibition of HBx protein (hepatitis B virus X protein) expression, FXR (farnesoid X receptor) inhibition, by confirming that the antiviral activity is shown, fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) provides a novel antiviral agent.

Description

Antiviral composition comprising fibroblast growth factor 11 as an active ingredient}

The present invention relates to an antiviral composition comprising fibroblast growth factor 11 as an active ingredient.

Hepatitis B virus (HBV) is a DNA virus belonging to the Hepadnaviridae family. It is known as one of the viruses that cause damage to people.

According to the World Health Organization, there are about 240 million chronic HBV infections worldwide, and between 500,000 and 700,000 people each year develop illnesses caused by hepatitis B virus infection. reported dead In Korea, 5-8% of adults are known to be HBV carriers, and 80% of chronic hepatitis patients, 65% of liver cirrhosis, and 70% of hepatocellular carcinoma patients in adults have been investigated to be associated with HBV infection.

HBV infection is the most important cause of chronic liver disease, and liver disease is considered one of the most important social problems in Korea. Therefore, appropriate antiviral treatment that can prevent progression to chronic liver disease or prevent new infection in already infected patients is essential.

Various types of treatments for HBV are being developed, and the annual market size is over 3.5 trillion won worldwide, the domestic market is over 200 billion won annually, and more than 1.5 trillion won in China is used annually. However, conventionally developed HBV therapeutics show excellent therapeutic effects from 1 to 5 years after the initial treatment, but as viral mutations occur, there is a problem in that the therapeutic effect is significantly reduced due to resistant viruses to the therapeutic agents.

Most of the conventionally developed HBV therapeutic agents work by targeting viral DNA or RNA polymerase, and studies have been reported that the mechanism of these therapeutic agents induces viral mutation. Therefore, there is a need for the development of an HBV therapeutic agent that operates with a new mechanism that does not induce viral mutation.

Korean Patent Publication No. 10-2020-0049728 (Announced on May 8, 2020)

The present invention relates to an antiviral composition comprising fibroblast growth factor 11 as an active ingredient, wherein fibroblast growth factor 11 (FGF11) is effective against hepatitis B virus (HBV) in liver cancer cells. fibroblast growth factor 11 (FGF11) as a new product by confirming that it exhibits antiviral activities such as inhibition of gene expression, inhibition of HBx protein (hepatitis B virus X protein) expression, and inhibition of farnesoid X receptor (FXR). It is provided as an antiviral agent.

The present invention provides an antiviral composition comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of viral infectious diseases comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving viral infection diseases comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

According to the present invention, unlike conventional antiviral agents targeting RNA polymerase, fibroblast growth factor 11 (FGF11) inhibits gene expression for hepatitis B virus (HBV), HBx Since it exhibits antiviral activity through mechanisms such as protein (hepatitis B virus X protein) expression inhibition and FXR (farnesoid X receptor) inhibition, it can be provided as a new antiviral agent as well as a combination administration with existing antiviral agents. can

1 is a diagram schematically illustrating the antiviral activity of fibroblast growth factor 11 (FGF11) against hepatitis B virus (HBV) in cells.
Figure 2 is a graph evaluating the effect of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) on gene expression of hepatitis B virus (HBV) in liver cancer cell lines.
Figure 3 is a graph evaluating the effect of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) on mRNA (A) and protein (B) expression of HBx protein (hepatitis B virus X protein) in liver cancer cell lines.
Figure 4 is a graph evaluating the expression change of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) according to the expression level of hepatitis B virus (hepatitis B virus, HBV) genes in liver cancer cell lines.
Figure 5 evaluates the effect of fibroblast growth factor 11 (FGF11) on the expression of hepatitis B virus (HBV) gene induced by FXR (farnesoid X receptor) in liver cancer cell lines. This is the result.
Figure 6 evaluates the effect of fibroblast growth factor 11 (FGF11) on the expression of the hepatitis B virus (HBV) gene induced by an activator of the farnesoid X receptor (FXR) in liver cancer cell lines. is a result
7 is a result of evaluating the effect of fibroblast growth factor 11 (FGF11) on the expression of farnesoid X receptor (FXR) in liver cancer cell lines.

The terms used in this specification have been selected from general terms that are currently widely used as much as possible while considering the functions in the present invention, but these may vary depending on the intention of a person skilled in the art, precedent, or the emergence of new technologies. In addition, in a specific case, there is also a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, not simply the name of the term.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in this application, it should not be interpreted in an ideal or excessively formal meaning. don't

Numerical ranges are inclusive of the values defined therein. Every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written. Every minimum numerical limitation given throughout this specification includes every higher numerical limitation, as if such higher numerical limitations were expressly written. Every numerical limitation given throughout this specification will include every better numerical range within the broader numerical range, as if the narrower numerical limitations were expressly written.

Hereinafter, the present invention will be described in more detail.

The inventors of the present invention found that among the fibroblast growth factor family, fibroblast growth factor 11 (FGF11) inhibits the expression of viral proteins related to hepatitis B virus (HBV) replication. By first clarifying what it does, the present invention is provided.

The present invention provides an antiviral composition comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

The fibroblast growth factor 11 is a factor related to lipid metabolism in hepatocytes and adipocytes, and its expression is increased in hepatocytes under low oxygen conditions.

The fibroblast growth factor 11 (FGF11) suppresses the promoter activity of the hepatitis B virus (HBV) gene, expresses the HBx protein (hepatitis B virus X protein) or the gene encoding it It suppresses the expression of mRNA and suppresses the expression of FXR (farnesoid X receptor).

Since the fibroblast growth factor 11 (FGF11) does not induce genetic modification of hepatitis B virus (HBV) and works by targeting replication-related proteins, it can be used in combination with existing antiviral agents. When administered, the antiviral effect can be excellent.

The antiviral composition may be administered in combination with an antiviral agent, and may be administered simultaneously, separately or sequentially with the antiviral agent. Antiviral drugs include lamivudine, telbivudine, clevudine, entecavir, adefovir, tenofovir disoproxil, tenofovir alafenamide ), and may be any one or more selected from the group consisting of besifovir, but is not limited thereto.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of viral infectious diseases comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

The viral infection disease is a disease caused by infection with hepatitis B virus (HBV).

The pharmaceutical composition may be formulated in the form of one or more external agents selected from the group consisting of creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents.

The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres. binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol; don't The pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.

The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the desired method, and is preferably administered orally. In addition, the dosage of the pharmaceutical composition of the present invention varies depending on the subject's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease, but is not limited thereto.

In addition, the present invention provides a health functional food composition for preventing or improving viral infection diseases comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

The present invention can be generally used as a commonly used food.

The food composition of the present invention can be used as a health functional food. The term "health functional food" refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional" refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.

The food composition of the present invention may include conventional food additives, and the suitability as the "food additive" is determined according to the general rules of the food additive code approved by the Ministry of Food and Drug Safety and general test methods, etc., unless otherwise specified. It is judged according to the specifications and standards for the item.

Items listed in the "Food Additive Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

The food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like.

For example, among the health functional foods in the form of capsules, hard capsules can be prepared by mixing and filling the composition according to the present invention with additives such as excipients in conventional hard capsules, and soft capsules can be prepared by filling the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

Definitions of terms for the excipients, binders, disintegrants, lubricants, corrigents, flavoring agents, etc. are described in literature known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the usual sense.

In the present invention, the term "prevention" refers to all activities that inhibit or delay a disease by administering the composition according to the present invention. In the present invention, the term "treatment" refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention. In the present invention, "improvement" means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1. Effect of fibroblast growth factor 11 on gene expression of hepatitis B virus in liver cancer cell lines

In order to confirm the antiviral effect of fibroblast growth factor 11 (FGF11), fibroblast growth factor 11 (fibroblast growth factor 11) on gene expression of hepatitis B virus (HBV) in liver cancer cell lines 11, FGF11) was evaluated for its antiviral activity.

1.3x HBV-luc was used to evaluate gene expression of hepatitis B virus (HBV). 1.3x HBV-luc is a plasmid containing the full-length sequence of hepatitis B virus (HBV) and a promoter and enhancer for viral gene expression in liver cancer cell lines. By measuring the level, the degree of expression of the viral gene can be evaluated.

Liver cancer cell line Hep3B cells (American Type Culture Collection, ATCC ) were used. For Hep3B cell culture, DMEM (Dulbecco's modified Eagle's medium, WELGENE) medium containing 10% FBS (fetal bovine serum, Gibco BRL) inactivated by heat treatment and penicillin and streptomycin (Gibco BRL) was used at 37°C and 5% CO. It was cultured under ₂ conditions. 100ng 1.3x HBV-luc vector was transfected (transfection), and 100ng, 200ng, 300ng, 400ng or 500ng of FGF11 (Korea Institute of Marine Science and Technology ) was additionally transfected into Hep3B cells, followed by luciferase The expression and enzyme activity values of measured.

A luciferase reporter assay method for measuring HBV promoter activity was performed as follows. Hep3B cells were seeded into a 24-well culture plate at 6x10 ⁵ wells/cells, and each well was transfected with 100ng, 200ng, 300ng, 400ng or 500ng of 100ng of 1.3x HBV-luc plasmid and FGF11 plasmid at each concentration. At this time, the pCMV/flag vector was used as a control to standardize the total amount of transfected plasmid DNA. Transfection was performed according to the manufacturer's protocol using jetPEI (PolyPlus Transfection) when the cells reached 70-80% of the total area of the culture vessel. After 24 hours of transfection, cells were recovered using cell lysis buffer (Promega) and performed according to the protocol of the device using a luminescence luminometer (Promega).

As shown in FIG. 2, it was found that the activity level of luciferase decreased as fibroblast growth factor 11 (FGF11) was treated with 1.3x HBV-luc-infected liver cancer cell lines, As the treatment concentration of fibroblast growth factor 11 (FGF11) increased, the activity level of luciferase was greatly reduced. The above results show that fibroblast growth factor 11 (FGF11) has an activity to inhibit gene expression of hepatitis B virus (HBV), and that fibroblast growth factor 11 (FGF11) 11, proves the antiviral effect of FGF11).

Example 2. Effect of fibroblast growth factor 11 on HBx protein expression

In order to analyze the mechanism of the antiviral effect of fibroblast growth factor 11 (FGF11) against hepatitis B virus (HBV), fibroblast growth factor 11 (fibroblast growth factor 11) in liver cancer cell lines was 11, FGF11) on the expression of HBx protein (hepatitis B virus X protein) was evaluated.

(1) RT-PCR

HBx protein (hepatitis B virus X protein) is a pathogenic protein specifically expressed in hepatitis B virus that induces various liver diseases and plays a major role in regulating gene replication and expression of hepatitis B virus (HBV). It is known to To evaluate the effect of fibroblast growth factor 11 (FGF11) on the expression of hepatitis B virus X protein, hepatitis B virus (HBV)-infected liver cancer cell lines were treated with fibroblast cells. Growth factor 11 (fibroblast growth factor 11, FGF11) was treated, and the mRNA expression level and protein level of HBx protein (hepatitis B virus X protein) were measured.

HepG2, a human liver cancer cell line, was purchased from the American Type Culture Collection (ATCC), and DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS (fetal bovine serum, Gibco BRL) inactivated by heat treatment and penicillin and streptomycin (Gibco BRL) was used. , WELGENE) medium was used and cultured under 37°C and 5% CO ₂ conditions. HepG2 cells were transfected using jetOPTIMUS (PolyPlus Transfection). After 24 hours of transfection, total RNA was extracted using Trizol reagent (Invitrogen) after dissolution according to the manufacturer's protocol. 0.5 μg of extracted RNA and M-MLV reverse transcriptase (Enzynomics) were used to prepare a reaction solution so that the final solution was 20 μl, and then cDNA was synthesized. The cDNA synthesized according to the above process was subjected to the following conditions: first denaturation at 95 ° C for 10 minutes, followed by denaturation at 95 ° C (30 seconds), primer binding (30 seconds), polymerization at 72 ° C (30 seconds), 25 cycles repeated; And the final extension process was carried out at 72 ° C. for 5 minutes to amplify. Information on primer sets used for PCR and respective binding temperatures are shown in Table 1 below. The results were confirmed through electrophoresis using a 1.2% agarose gel.

Real-time quantitative PCR amplification was performed using the StepONEt ^TM Real-time PCR System using TOPreal qPCR × 2 PreMIX with SYBR green (Enzynomics). Relative mRNA quantitative analysis was performed using the ΔΔCt method, and the results were expressed as n-fold differences relative to the calibrator (RQ=2 ^-ΔΔCt ).

Primer base sequence sequence number bonding temperature HBx Forward 5'-ATGGCTGCTAGGCTGTTGCTGC-3' One 58℃ Reverse 5'- ACGGTGGTCTCCATGCGACG-3' 2 FGF11 Forward 5'-CCAAGTCCCTTTGCCAGAAGC-3' 3 56℃ Reverse 5'-CCATGTAGTGACCCAGCTTGG-3' 4 hGAPDH Forward 5'-GTGGTCTCCTTCTGACTTCAAC-3' 5 56℃ Reverse 5'-TCTCTTCCTCTTGTGCTCTTG-3' 6

(2) Western blot

Hep3B cells were transfected using jetPEI (PolyPlus Transfection). Cells were collected 24 hours after transfection, and cells were lysed using lysis buffer (150 mM NaCl, 1% NP-40, 1 mM EDTA, 50 mM Tris pH 7.5, protease inhibitor and 1 mM PMSF). The protein concentration in the cell lysate was measured using Bradford reagent (Bio-Rad). 30 μg of protein was subjected to SDS-PAGE using a 12-15% polyacrylamide gel and sorted by size. Next, the proteins in the polyacrylamide gel were transferred onto a PVDF membrane through a transfer process, and then treated with primary antibodies anti-actin (purchased from Sigma) and anti-flag (purchased from Cell signaling) diluted at 1:2000. reacted afterwards. Then, the expression level of the target protein was confirmed by processing the secondary antibody against the above primary antibody.

As shown in FIG. 3, it was found that the mRNA and protein expression levels of the hepatitis B virus X protein decreased as the liver cancer cell line was treated with fibroblast growth factor 11 (FGF11). The above results show that fibroblast growth factor 11 (FGF11) inhibits the expression of HBx protein (hepatitis B virus X protein), and fibroblast growth factor 11 against pathogenic viruses , FGF11) to demonstrate the antiviral effect.

Example 3. Fibroblast growth factor 11 expression change according to hepatitis B virus gene expression in liver cancer cell line

Changes in the expression of fibroblast growth factor 11 (FGF11) according to the degree of hepatitis B virus (HBV) infection were evaluated.

In order to evaluate the expression level of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11), a plasmid containing the promoter and luciferase reporter gene of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) was used in liver cancer cell lines. Hep3B cells were transfected.

The FGF11-luc plasmid used in the present invention was provided and used by the Korea Institute of Ocean Science & Technology (KIOST). The FGF11-luc plasmid was constructed using the luciferase pGL3 promoter vector (Promega) by referring to the FGF11 promoter information represented by the nucleotide sequence of SEQ ID NO: 9.

Transfected Hep3B cells were treated with 100ng, 200ng, 300ng, 400ng or 500ng of HBV replicon (1.3x HBV-luc) containing total genomic DNA of hepatitis B virus (HBV), and FGF11 A luciferase reporter assay was performed to determine the level of promoter activity in the following manner. Hep3B cells were seeded into a 24-well culture plate at 6x10 ⁵ wells/cells, and each well was transfected with 100ng, 200ng, 300ng, 400ng or 500ng of FGF11-luc plasmid and HBV replicon at each concentration. After 24 hours of transfection, cells were recovered using cell lysis buffer (Promega), and performed using a luminescence luminometer (Promega) according to the protocol of the device.

As shown in FIG. 4, it was found that the activity level of luciferase increased as the replicon of hepatitis B virus (HBV) was treated, and the result showed that the hepatitis B virus (hepatitis B It shows that as the infection of virus (HBV) increases, the expression of fibroblast growth factor 11 (FGF11) increases to suppress the infection of the virus in hepatocytes.

Example 4. Effect of fibroblast growth factor 11 on FXR activity

To analyze the antiviral mechanism of fibroblast growth factor 11 (FGF11) against hepatitis B virus (HBV), fibroblast growth factor 11 (FGF11) was FXR The effect on the activity of the farnesoid X receptor was evaluated.

FXR (farnesoid X receptor) is a nuclear receptor factor in hepatocytes and is known as a factor that activates viral gene expression when hepatocytes are infected with hepatitis B virus (HBV).

Fibroblast growth factor 11( The effect of fibroblast growth factor 11, FGF11) was measured.

Except for the treatment of FXR (farnesoid X receptor) α1 (UCLA Medicine), gene expression changes of hepatitis B virus (HBV) in liver cancer cell lines proceeded in the same manner as in Example 1, and HBx protein ( hepatitis B virus X protein) was performed in the same manner as in Example 2.

(1) RT-PCR

HepG2 cells were transfected using jetOPTIMUS (PolyPlus Transfection). After 24 hours of transfection, total RNA was extracted using Trizol reagent (Invitrogen) according to the manufacturer's protocol. 0.5 μg of extracted RNA and M-MLV reverse transcriptase (Enzynomics) were used to prepare a reaction solution so that the final solution was 20 μl, and then cDNA was synthesized. The cDNA synthesized according to the above process was performed under the following conditions: first denaturation at 95 ° C for 10 minutes, then denaturation at 95 ° C (30 seconds), primer binding temperature (table below) (30 seconds), polymerization at 72 ° C (30 seconds) seconds), 25 cycles were repeated, and the final extension process was carried out at 72° C. for 5 minutes to amplify. Information on primer sets used for PCR and respective binding temperatures are shown in Table 2 below. The results were confirmed through electrophoresis using a 1.2% agarose gel.

Real-time quantitative PCR amplification was performed using the StepONEt ^TM Real-time PCR System using TOPreal qPCR × 2 PreMIX with SYBR green (Enzynomics). Relative mRNA quantitative analysis was performed using the ΔΔCt method, and the results were expressed as n-fold differences relative to the calibrator (RQ=2 ^-ΔΔCt ).

Primer base sequence sequence number bonding temperature HBx Forward 5'-ATGGCTGCTAGGCTGTTGCTGC-3' One 58℃ Reverse 5'-ACGGTGGTCTCCATGCGACG-3' 2 FGF11 Forward 5'-CCAAGTCCCTTTGCCAGAAGC-3' 3 56℃ Reverse 5'-CCATGTAGTGACCCAGCTTGG-3' 4 hGAPDH Forward 5'-GTGGTCTCCTTCTGACTTCAAC-3' 5 56℃ Reverse 5'-TCTCTTCCTCTTGTGCTCTTG-3' 6 FXRa1 Forward 5'-AGGGGATGAGCTGTGTGTTG-3' 7 58℃ Reverse 5'-CCTGTATACATACATTCAGCCAAC-3' 8

(2) Western blot

HepG2 cells were transfected using jetOPTIMUS (PolyPlus Transfection). Cells were collected 24 hours after transfection, and cells were lysed using lysis buffer (150 mM NaCl, 1% NP-40, 1 mM EDTA, 50 mM Tris pH 7.5, protease inhibitor and 1 mM PMSF). The protein concentration in the cell lysate was measured using Bradford reagent (Bio-Rad). 30 μg of protein was subjected to SDS-PAGE using a 10-15% polyacrylamide gel and sorted by size. Next, the proteins in the polyacrylamide gel were transferred onto a PVDF membrane through a transfer process, and the primary antibodies anti-actin (Sigma) diluted at 1:2000, anti-flag (Cell signaling), anti-FXR ( Santa Cruz Biotechnology) and then reacted. Then, the expression level of the target protein was confirmed by processing the secondary antibody against the above primary antibody.

As shown in FIG. 5, as liver cancer cells are treated with FXR (farnesoid X receptor), hepatitis B virus (HBV) gene expression increases and HBx protein (hepatitis B virus X protein) expression increases However, as fibroblast growth factor 11 (FGF11) was treated, hepatitis B virus (HBV) gene expression and HBx protein (hepatitis B virus X protein) increased with FXR (farnesoid X receptor) ) was found to decrease.

In addition, as GW4064, an activator that binds to FXR (farnesoid X receptor), is additionally treated, fibroblast growth factor 11 (FGF11) induces hepatitis B virus (HBV) gene expression in liver cancer cell lines. Changes in the expression of HBx protein (hepatitis B virus X protein) were evaluated.

As shown in Figure 6, the gene expression of hepatitis B virus (HBV) and the expression of HBx protein (hepatitis B virus X protein) increased by FXR (farnesoid X receptor) and GW4064 were fibroblast growth factor 11 (fibroblast growth factor 11, FGF11).

The above results show that fibroblast growth factor 11 (FGF11) exhibits antiviral activity by targeting farnesoid X receptor (FXR).

Example 5. Effect of fibroblast growth factor 11 on FXR gene expression

In order to analyze the antiviral mechanism of fibroblast growth factor 11 (FGF11) against hepatitis B virus (HBV), fibroblast growth factor 11 (FGF11) was FXR The effect on the expression of (farnesoid X receptor) was evaluated. The method for measuring the expression level of FXR (farnesoid X receptor) was carried out in the same manner as in Example 4.

As shown in FIG. 7, the expression of farnesoid X receptor (FXR) was found to decrease as fibroblast growth factor 11 (FGF11) was treated.

The above results show that fibroblast growth factor 11 (FGF11) reduces the activity and production of farnesoid X receptor (FXR) transcription factor that increases the replication and expression of hepatitis B virus (HBV). , inhibits the expression of hepatitis B virus (HBV), which makes fibroblast growth factor 11 (FGF11) a new antiviral agent against hepatitis B virus (HBV) prove that it can be used.

Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

<110> Pusan National Uni. <120> Antiviral composition comprising fibroblast growth factor 11 as an active ingredient <130> ADP-2020-0223 <160> 9 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> artificial sequence <220> <223> HBx-f <400> 1 atggctgcta ggctgtgctg c 21 <210> 2 <211> 20 <212> DNA <213> artificial sequence <220> <223> HBx-r <400> 2 acggtggtct ccatgcgacg 20 <210> 3 <211> 21 <212> DNA <213> artificial sequence <220> <223> FGF11-f <400> 3 ccaagtccct ttgccagaag c 21 <210> 4 <211> 21 <212> DNA <213> artificial sequence <220> <223> FGF11-r <400> 4 ccatgtagtg acccagcttg g 21 <210> 5 <211> 21 <212> DNA <213> artificial sequence <220> <223> hGAPDH-f <400> 5 gtggtctcct ctgacttcaa c 21 <210> 6 <211> 21 <212> DNA <213> artificial sequence <220> <223> hGAPDH-r <400> 6 tctcttcctc ttgtgctctt g 21 <210> 7 <211> 20 <212> DNA <213> artificial sequence <220> <223> FXRa1-f <400> 7 aggggatgag ctgtgtgttg 20 <210> 8 <211> 24 <212> DNA <213> artificial sequence <220> <223> FXRa1-r <400> 8 cctgtataca tacattcagc caac 24 <210> 9 <211> 1800 <212> DNA <213> artificial sequence <220> <223> FGF11 <400> 9 agcgtccgcc attccatctc ccttaggccc aacctctcct tcctacccag gggtgcgttc 60 aggagggcctt cctgtccctc ctcctctgaa ggctgagacg tgcggccttg cctcggcttt 120 gcctgcttgt aagacacagt ccgcactctc tacctccaag gaaccccggg gttcctcctg 180 cccggcttca ccctcctatt tcccagcgtc tccgccccct gccccgcccc cgggccggct 240 ctccgaggag ggggaactgc tgtcgccgcc gccgccgccg cctcagcttc ccacagccgc 300 tgccgctgcc gccgctctgc ctggtccagc caccggccca gtgctctgac ttcgccggac 360 cggggagctc tgcagagaca ccctcactcc ttcccgaggg tccgggacgc ctagccgggc 420 gtgggggggaa gctccgtttg cgggggacag ggagggagga gcctggagcc gaagccagcg 480 ccacccgtcg ccggatcaac aggacaggac aggttgaggg gcgtcgggaa aaggaagagg 540 gggtgctata ggcaatcccg gggaggacga ggaagccctg tagcaggaag ctgtgatccg 600 ctctgacttg tagggggcct gttcacatct ggggactggt gacaacgtgg gtgcactcct 660 720 gaaaatttaa ggggcagcca ggggagaaaa aaacaaaaac aaaaacaaac acggatgctc 780 ctcattttga ggtggcggaa ggccctccag tcaactacct ggtgtatcca ctaggaaggg 840 tggatttgga ggtgacttca aaaggagcgg agggtgtgaa ggtgaagaaa gggtcttggc 900 cgctagaatt ctatgctact agacatgggg gggacttggt gaaaaaggta ttatccagcc 960 agagggtctg ggagccctgt cttactgaac ctgggcaacc tggatattct gagacatatt 1020 ttggggggat ttcagtgaaa aaagtggggg atcccctcca tttagagtgt agcaaaggaa 1080 aaaacaccaa ggttgggttc cttcctgaca ttggcagtgc cccagtaggg gtgggatgag 1140 cgaatattcc caaagctaaa gtcccacacc ctgtagatta caagagtgga tttggcagga 1200 gtgtgcccca aaatacagtg gaaaggtgcc tgaagatatt taaaccacgt cttggaaatt 1260 tagtgggtct tggctttggg ataggtgaag tgaggacaga cactggagag gagggaaagg 1320 ggacgttttc aataggaggc aaaactcgag ggtgggatcc actgaggagt acataggctg 1380 ctggatctgg tggagccagc actgggccca cgggtggtaa ctggctgctg tggaggggggg 1440 tacgtgaggg ggggggtctg gggcttatcc tcaggtcctg tgggtggggc agcgagtcgg 1500 ggcctgagcg tcaagagcat gccctagtga gcgggctcct ctgggggagc ccagcgcgct 1560 ccgggcgcct gccggtttgg gggtgtctcc tcccggggcg ctatggcggc gctggccagt 1620 agcctgatcc ggcagaagcg ggaggtccgc gagcccgggg gcagccggcc ggtgtcggcg 1680 cagcggcgcg tgtgtccccg cggcaccaag tccctttgcc agaagcagct cctcatcctg 1740 ctgtccaagg tgcgactgtg cggggggcgg cccgcgcggc cggaccgcgg cccgggtgag 1800 1800

Claims (10)

A pharmaceutical composition for preventing or treating hepatitis B virus (HBV) infectious diseases, comprising fibroblast growth factor 11 (FGF11) as an active ingredient. The pharmaceutical composition according to claim 1, wherein the fibroblast growth factor 11 (FGF11) inhibits the promoter activity of the hepatitis B virus (HBV) gene. The pharmaceutical composition according to claim 1, wherein the fibroblast growth factor 11 (FGF11) inhibits the expression of hepatitis B virus X protein or the mRNA expression of a gene encoding the same. . The pharmaceutical composition according to claim 1, wherein the fibroblast growth factor 11 (FGF11) inhibits the expression of farnesoid X receptor (FXR). The pharmaceutical composition according to claim 1, wherein the antiviral composition is administered in combination with an antiviral agent. The pharmaceutical composition according to claim 5, wherein the antiviral composition is administered simultaneously with the antiviral agent, separately or sequentially. The method of claim 5, wherein the antiviral agent is lamivudine, telbivudine, clevudine, entecavir, adefovir, tenofovir disoproxil, tenofor A pharmaceutical composition, characterized in that at least one selected from the group consisting of tenofovir alafenamide and besifovir. delete delete A health functional food composition for preventing or improving hepatitis B virus (HBV) infectious disease, comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

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