KR20220023204A - 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. It has been confirmed that fibroblast growth factor 11 (FGF11) exhibits antiviral activity such as inhibition of gene expression for hepatitis B virus (HBV) in liver cancer cells, inhibition of hepatitis B virus X protein expression, and inhibition of farnesoid X receptor (FXR), thereby providing fibroblast growth factor 11 (FGF11) as a new antiviral agent.

Description

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

According to the World Health Organization, there are approximately 240 million chronic HBV infections worldwide, and between 500,000 and 700,000 people are diagnosed with hepatitis B virus infection each year. reported to have died. In Korea, 5-8% of adults are HBV carriers, and 80% of chronic hepatitis patients, 65% of liver cirrhosis patients, and 70% of hepatocellular carcinoma patients in adults are related to HBV infection.

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

Various types of therapeutic agents for HBV are being developed, and the annual global market is more than 3.5 trillion won, the domestic market size is more than 200 billion won, and in China, more than 1.5 trillion won of therapeutic agents are being used annually. However, the conventionally developed HBV therapeutic agent showed excellent therapeutic effect from 1 to 5 years from the start of the initial treatment, but there is a problem in that the therapeutic effect is significantly reduced due to the virus resistant to the therapeutic agent as the virus mutation occurs.

Most HBV therapeutics previously developed work by targeting viral DNA or RNA polymerase, and studies have been reported that the mechanism of these therapeutics induces viral mutation. Therefore, there is a demand for the development of a therapeutic agent for HBV that works with a novel mechanism that does not induce viral mutation.

Korean Patent Application Laid-Open No. 10-2020-0049728 (2020. 05. 08. Announcement)

The present invention relates to an antiviral composition comprising fibroblast growth factor 11 as an active ingredient, wherein fibroblast growth factor 11 (FGF11) is a hepatitis B virus (HBV) in liver cancer cells. Fibroblast growth factor 11 (FGF11) was newly developed by confirming that it exhibits antiviral activity such as suppression of gene expression for 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 preventing or treating viral infections 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 disease comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

According to the present invention, fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) inhibits gene expression for hepatitis B virus (HBV), unlike conventional antiviral agents that target RNA polymerase, HBx Because it exhibits antiviral activity through mechanisms such as suppression of hepatitis B virus X protein expression and inhibition of FXR (farnesoid X receptor), 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 the gene expression of hepatitis B virus (hepatitis B virus, HBV) in a liver cancer cell line.
Figure 3 is a graph evaluating the effect of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) on the mRNA (A) and protein (B) expression of HBx protein (hepatitis B virus X protein) in a liver cancer cell line.
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 the hepatitis B virus (hepatitis B virus, HBV) gene in a liver cancer cell line.
5 is a liver cancer cell line fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) FXR (farnesoid X receptor) induced by hepatitis B virus (hepatitis B virus, HBV) to evaluate the effect on the gene expression. is the result
6 is a liver cancer cell line fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) FXR (farnesoid X receptor) induced by an activator induced hepatitis B virus (hepatitis B virus, HBV) to evaluate the effect on the gene expression is a result
7 is a result of evaluating the effect of fibroblast growth factor 11 (FGF11) on the expression of FXR (farnesoid X receptor) in a liver cancer cell line.

The terms used in this specification have been selected as currently widely used general terms as possible while considering the functions in the present invention, but these may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like. In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding 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, rather than the name of a simple term.

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

Numerical ranges are inclusive of the values defined in that range. Every maximum numerical limitation given throughout this specification includes all lower numerical limitations as if the lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes all higher numerical limitations as if the higher numerical limitation were expressly written. Any numerical limitation given throughout this specification shall include all numerical ranges within the broader numerical range, as if the narrower numerical limitation were expressly written down.

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

The inventors of the present invention have found that among the fibroblast growth factor families, fibroblast growth factor 11 (FGF11) suppresses the expression of viral proteins related to hepatitis B virus (HBV) replication. By first finding out what to do, 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 an interest related to lipid metabolism in hepatocytes and adipocytes, and its expression is increased in hepatocytes under low oxygen conditions.

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

Since the fibroblast growth factor 11 (FGF11) does not induce genome modification of hepatitis B virus (HBV) and works by targeting a replication-related protein, it is used in combination with conventional antiviral agents When administered, the antiviral effect may be excellent.

The antiviral composition may be administered in combination with an antiviral agent, and may be administered simultaneously with the antiviral agent (simultaneous), separately (separate) or sequentially (seqeuntial). Antiviral agents 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 preventing or treating viral infections comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

The viral infection 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 preparations selected from the group consisting of creams, gels, patches, sprays, ointments, warning agents, lotions, liniments, pastas, and cataplasmas.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier and diluent for formulation. The pharmaceutically acceptable carriers and diluents include starch, sugar, and excipients such as mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethyl cellulose, hydroxypropyl cellulose, gelatin, alginate, and polyvinyl blood binders such as rolidone, lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol does not 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 (eg, intravenously, subcutaneously, intraperitoneally or topically) according to a 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 status, 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 disease comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

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

The food composition of the present invention can be used as a health functional food. The term "health functional food" means a food manufactured and processed using raw materials or ingredients useful for the human body in accordance with the Health Functional Food Act, and "functionality" refers to the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological effects.

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

The items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; Mixed preparations such as sodium L-glutamate preparation, noodle-added alkali agent, preservative agent, and tar color agent can be mentioned.

The food composition of the present invention may be manufactured 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 a conventional hard capsule with the composition according to the present invention with additives such as excipients, and the soft capsule is a composition according to the present invention. It can be manufactured by mixing with additives such as excipients and filling in capsule bases such as gelatin. The soft capsule formulation may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

The term definitions for the excipients, binders, disintegrants, lubricants, flavoring agents, and the like are described in documents known in the art and include those having the same or similar functions. The type of food is not particularly limited, and includes all health functional foods in the ordinary sense.

In the present invention, the term “prevention” refers to any act of inhibiting or delaying a disease by administering the composition according to the present invention. In the present invention, the term “treatment” refers to any action that improves or beneficially changes the symptoms of a disease by administration of the composition according to the present invention. In the present invention, "improvement" refers to any action that improves the bad state of a disease by administering or ingesting the composition of the present invention to an individual.

Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the content of the present invention, and 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 of ordinary skill in the art.

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

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

To evaluate the gene expression of hepatitis B virus (HBV), 1.3x HBV-luc was used. 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 a liver cancer cell line. The activity of luciferase By measuring the level, the expression level of the viral gene can be assessed.

Hep3B cells (American Type Culture Collection, ATCC ) , a liver cancer cell line, were used. For Hep3B cell culture, using 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) at 37°C, 5% CO It was cultured under ₂ conditions. 100ng 1.3x HBV-luc vector was transfected (transfection), and FGF11 (Korea Institute of Ocean Science and Technology ) was additionally transfected with 100ng, 200ng, 300ng, 400ng or 500ng Hep3B cells and then luciferase values of expression and enzymatic activity of measured.

The luciferase reporter assay method for measuring the HBV promoter activity value was performed as follows. Hep3B cells were seeded in a 24-well culture plate by 6x10 ⁵ wells/cells, and 100ng, 200ng, 300ng, 400ng or 500ng of 100ng of 1.3x HBV-luc plasmid and FGF11 plasmid were transfected into each well by concentration. At this time, pCMV/flag vector was used as a control to normalize 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. 24 hours after transfection, cells were recovered using cell lyis buffer (Promega), and a luminescence luminometer (Promega) was used according to the protocol of the device.

As shown in FIG. 2, as the liver cancer cell line infected with 1.3x HBV-luc was treated with fibroblast growth factor 11 (FGF11), the activity level of luciferase was decreased. As the treatment concentration of fibroblast growth factor 11 (FGF11) increased, the activity level of luciferase was significantly reduced. The result shows that fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) has an activity to inhibit gene expression of hepatitis B virus (HBV), fibroblast growth factor 11 (fibroblast growth factor) 11, demonstrating the antiviral effect of FGF11).

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

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

(1) RT-PCR

HBx protein (hepatitis B virus X protein) is a pathogenic protein that appears specifically in hepatitis B virus that induces various liver diseases. It plays a major role in regulating gene replication and expression of hepatitis B virus (HBV). is known to do To evaluate the effect of fibroblast growth factor 11 (FGF11) on HBx protein (hepatitis B virus X protein) expression, hepatitis B virus (HBV)-infected liver cancer cell lines were treated with fibroblasts. 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, penicillin and streptomycin (Gibco BRL) , WELGENE) medium was used at 37° C., and cultured under 5% CO ₂ conditions. HepG2 cells were transfected using jetOPTIMUS (PolyPlus Transfection). 24 hours after transfection, total RNA was extracted after dissolution using Trizol reagent (Invitrogen) according to the manufacturer's protocol. Using 0.5 μg of extracted RNA and M-MLV reverse transcriptase (Enzynomics), a reaction solution was prepared so that the final solution was 20 μl, and cDNA was synthesized. The cDNA synthesized according to the above procedure was performed under the following conditions: primary denaturation at 95°C for 10 minutes, denaturation at 95°C (30 seconds), primer binding (30 seconds), polymerization at 72°C (30 seconds) 25 cycles were repeated; And the final extension process was carried out at 72 ℃ for 5 minutes to amplify. Information on primer sets used for PCR and their respective binding temperatures are shown in Table 1 below. The results were confirmed through electrophoresis using 1.2% agarose gel.

The real-time quantitative PCR amplification process was performed using the StepONEt ^TM Real-time PCR System using TOPreal qPCR × 2 PreMIX with SYBR green (Enzynomics). The ΔΔCt method was used for relative mRNA quantitative analysis, and the result was expressed as an n-fold difference relative to the calibrator (RQ=2 ^-ΔΔCt ).

Primer base sequence SEQ ID NO: bonding temperature HBx Forward 5’-ATGGCTGCTAGGCTGTGCTGC-3’ One 58℃ Reverse 5’-ACGGTGGTCTCCATGCGACG-3’ 2 FGF11 Forward 5’-CCAAGTCCCTTTGCCAGAAGC-3’ 3 56℃ Reverse 5’-CCATGTAGTGACCCAGCTTGG-3’ 4 hGAPDH Forward 5’-GTGGTCTCCTCTGACTTCAAC-3’ 5 56℃ Reverse 5’-TCTCTTCCTCTTGTGCTCTTG-3’ 6

(2) Western blot

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

As shown in FIG. 3 , as the liver cancer cell line was treated with fibroblast growth factor 11 (FGF11), mRNA and protein expression levels of HBx protein (hepatitis B virus X protein) were decreased. The result shows that fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) suppresses the expression of HBx protein (hepatitis B virus X protein) along with fibroblast growth factor 11 for pathogenic viruses , demonstrating the antiviral effect of FGF11).

Example 3. Changes in expression of fibroblast growth factor 11 according to hepatitis B virus gene expression in liver cancer cell lines

The expression change of fibroblast growth factor 11 (FGF11) according to the degree of infection with hepatitis B virus (HBV) was evaluated.

To evaluate the expression level of fibroblast growth factor 11 (fibroblast growth factor 11, FGF11), a plasmid containing a fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) promoter and a luciferase reporter gene, a liver cancer cell line 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) with reference to the FGF11 promoter information expressed by the nucleotide sequence of SEQ ID NO: 9.

The transfected Hep3B cells were treated with 100ng, 200ng, 300ng, 400ng or 500ng of an HBV replicon (1.3x HBV-luc) containing whole genomic DNA of hepatitis B virus (HBV), and FGF11 The level of promoter activity was analyzed by luciferase reporter in the following way. Hep3B cells were aliquoted in a 24-well culture plate by 6x10 ⁵ wells/cells, and 100ng, 200ng, 300ng, 400ng or 500ng of 100ng of FGF11-luc plasmid and HBV replicon were transfected into each well by concentration. 24 hours after transfection, cells were recovered using cell lyis buffer (Promega), and a luminescence luminometer (Promega) was used 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 was hepatitis B virus (hepatitis B). It shows that the expression of fibroblast growth factor 11 (FGF11) increases to suppress viral infection in hepatocytes as the infection of the virus, HBV increases.

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 (farnesoid X receptor) effect on the activity was evaluated.

FXR (farnesoid X receptor) is a nuclear receptor factor in hepatocytes, and it is known as a factor activating viral gene expression when hepatitis B virus (HBV) is infected.

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

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

(1) RT-PCR

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

The real-time quantitative PCR amplification process was performed using the StepONEt ^TM Real-time PCR System using TOPreal qPCR × 2 PreMIX with SYBR green (Enzynomics). The ΔΔCt method was used for relative mRNA quantitative analysis, and the result was expressed as an n-fold difference relative to the calibrator (RQ=2 ^-ΔΔCt ).

Primer base sequence SEQ ID NO: bonding temperature HBx Forward 5’-ATGGCTGCTAGGCTGTGCTGC-3’ One 58℃ Reverse 5’-ACGGTGGTCTCCATGCGACG-3’ 2 FGF11 Forward 5’-CCAAGTCCCTTTGCCAGAAGC-3’ 3 56℃ Reverse 5’-CCATGTAGTGACCCAGCTTGG-3’ 4 hGAPDH Forward 5’-GTGGTCTCCTCTGACTTCAAC-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 harvested 24 hours after transfection, and cells were lysed using a lysis buffer (150 mM NaCl, 1% NP-40, 1 mM EDTA, 50 mM Tris pH 7.5, protease inhibitor and 1 mM PMSF). The concentration of the protein in the cell lysate was measured using Bradford reagent (Bio-Rad). Using a 10-15% polyacrylamide gel, 30 μg of protein was subjected to SDS-PAGE and sorted by size. Next, the proteins in the polyacrylamide gel are transferred onto the PVDF membrane through a transfer process, and then the primary antibodies anti-actin (Sigma), anti-flag (Cell signaling), and anti-FXR ( Santa Cruz Biotechnology) was treated and then reacted. And 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 the liver cancer cells are treated with FXR (farnesoid X receptor), the gene expression of hepatitis B virus (HBV) increases, and the expression of the HBx protein (hepatitis B virus X protein) increases. However, with fibroblast growth factor 11 (FGF11) treatment, the gene expression of hepatitis B virus (HBV) increased by FXR (farnesoid X receptor) and HBx protein (hepatitis B virus X protein) ) was found to decrease.

In addition, as GW4064, an activator that binds to FXR (farnesoid X receptor), was additionally treated, fibroblast growth factor 11 (FGF11) was expressed in 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 FIG. 6 , the gene expression of hepatitis B virus (HBV) increased by FXR (farnesoid X receptor) and GW4064 and the expression of HBx protein (hepatitis B virus X protein) were increased by fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) was shown to decrease.

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

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) is FXR (farnesoid X receptor) was evaluated for its effect on expression. A method of measuring the expression level of FXR (farnesoid X receptor) was carried out in the same manner as in Example 4.

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

The results show that fibroblast growth factor 11 (FGF11) reduces the activity and production of FXR (farnesoid X receptor) transcription factors that increase the replication and expression of hepatitis B virus (HBV). , suppresses the expression of hepatitis B virus (HBV), which uses fibroblast growth factor 11 (FGF11) as a novel antiviral agent for hepatitis B virus (HBV). prove that it can be used.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, 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 aggaggcctt 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 gtggggggaa 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 ctcttggggg gctggttaga ttggagggcg tgggatctaa acagcaccag ctgtccccaa 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 tggaggggg 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)

An antiviral composition comprising fibroblast growth factor 11 (FGF11) as an active ingredient. The antiviral composition of claim 1, wherein the fibroblast growth factor 11 (FGF11) inhibits the promoter activity of the hepatitis B virus (HBV) gene. The antiviral composition according to claim 1, wherein the fibroblast growth factor 11 (FGF11) inhibits the expression of hepatitis B virus X protein (HBx) or the mRNA expression of a gene encoding the same. . According to claim 1, wherein the fibroblast growth factor 11 (fibroblast growth factor 11, FGF11) is an antiviral composition characterized in that it inhibits the expression of FXR (farnesoid X receptor). The antiviral composition according to claim 1, wherein the antiviral composition is administered in combination with an antiviral agent. The antiviral composition according to claim 5, wherein the antiviral composition is administered simultaneously with the antiviral agent (simultaneous), separately (separate) or sequentially (seqeuntial). The method of claim 5, wherein the antiviral agent is lamivudine, telbivudine, clevudine, entecavir, adefovir, tenofovir disoproxil, tenofo Antiviral composition, characterized in that at least one selected from the group consisting of vir alafenamide (tenofovir alafenamide), and besifovir (besifovir). A pharmaceutical composition for preventing or treating viral infections comprising fibroblast growth factor 11 (FGF11) as an active ingredient. The pharmaceutical composition according to claim 8, wherein the viral infection is caused by infection with hepatitis B virus (HBV). A health functional food composition for preventing or improving viral infection disease comprising fibroblast growth factor 11 (FGF11) as an active ingredient.

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