Composition for inhibiting hepatitis b virus (HBV) proliferation

Abstract

The present application relates to a composition for inhibiting HBV proliferation, the composition comprising a compound represented by chemical formula 1 below as an active ingredient. [Chemical formula 1] (In chemical formula 1, R is a substitutable C3-C20 cycloalkyl group or a substitutable C6-C20 aryl group, and the substitution is substitution by oxygen, nitrogen, sulfur, hydroxy, linear or branched C1-C6 alkyl, a C3-C20 cycloalkyl group, C6-C20 aryl, a halogen, or a combination thereof.)

Description

Composition for inhibiting the proliferation of Hepatitis B virus (HBV)

[0001] The present invention relates to a composition for inhibiting the proliferation of hepatitis B virus (HBV).

[0002] Hepatitis B virus (HBV) is one of the major pathogens causing serious health problems worldwide. According to the World Health Organization (WHO), approximately 250 million people are infected with chronic hepatitis B, and about 900,000 people die each year from hepatitis B-related complications.

[0003] Currently, nucleoside / nucleotide analogs and interferon-alpha are primarily used to treat hepatitis B. However, these treatments have drawbacks, such as difficulty in achieving complete viral elimination, the development of resistance with long-term administration, and side effects. Therefore, there is an urgent need for the development of anti-HBV drugs with novel mechanisms of action.

[0004] Recent studies have revealed that the sirtuin protein family plays a crucial role in the HBV life cycle. In particular, Sirtuin 2 (SIRT2) is known to act as an important regulator in the process of HBV replication. Consequently, interest in developing inhibitors targeting SIRT2 is growing. AGK2 is one of the well-known SIRT2 inhibitors that demonstrated an inhibitory effect on HBV proliferation in in vitro experiments; however, its clinical application has been limited due to low selectivity and side effects.

[0005] Therefore, there is a need for research to improve the efficacy of inhibiting HBV proliferation and reduce side effects through the development of new variants based on the structure of AGK2.

[0006] Korean Published Patent No. 10-2020-0123392, which is the technical background of the present invention, relates to a composition for inhibiting the proliferation of the hepatitis B virus and a method thereof.

[0007] The present invention aims to solve the problems of the aforementioned prior art by providing a composition for inhibiting HBV proliferation comprising a compound represented by the following chemical formula 1 as an active ingredient:

[0008] [Chemical Formula 1]

[0009]

[0010] (In the above chemical formula 1,

[0011] R is a substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0012] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl, halogen, or a combination thereof).

[0013] In addition, a pharmaceutical composition for preventing or treating HBV infection is provided, comprising the above-mentioned composition for inhibiting HBV proliferation.

[0014] In addition, a health functional food for preventing or improving HBV infection is provided, comprising a compound represented by the above chemical formula 1 as an active ingredient.

[0015] However, the technical problems that the embodiments of the present invention aim to solve are not limited to the technical problems described above, and other technical problems may exist.

[0016] As a technical means for achieving the above-mentioned technical problem, the first aspect of the present invention provides a composition for inhibiting HBV proliferation comprising a compound represented by the following chemical formula 1 as an active ingredient:

[0017] [Chemical Formula 1]

[0018]

[0019] (In the above chemical formula 1,

[0020] R is a substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0021] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl, halogen, or a combination thereof).

[0022] According to one embodiment of the present invention, the compound represented by Formula 1 may include any one of the following compounds, but is not limited thereto:

[0023] ;

[0024] .

[0025] According to one embodiment of the present invention, HBc (Hepatitis B core) protein expression in HepG2 cells may be inhibited by a compound represented by the above chemical formula 1, but is not limited thereto.

[0026] According to one embodiment of the present invention, the formation of a nucleocapsid in HepG2 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0027] According to one embodiment of the present invention, HBV DNA synthesis in HepG2 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0028] According to one embodiment of the present invention, HBV DNA synthesis in Huh7 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0029] In addition, the second aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to the first aspect of the present invention.

[0030] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0031] According to one embodiment of the present invention, the carrier may comprise, but is not limited to, a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

[0032] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally include a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof, but is not limited thereto.

[0033] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may be administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof, but is not limited thereto.

[0034] In addition, the third aspect of the present invention provides a health functional food for preventing or improving HBV infection, comprising a compound represented by Chemical Formula 1 of the first aspect of the present invention as an active ingredient.

[0035] The means for solving the problem described above are merely exemplary and should not be interpreted as intended to limit the present invention. In addition to the exemplary embodiments described above, additional embodiments may exist in the drawings and the detailed description of the invention.

[0036] The composition for inhibiting HBV proliferation according to the present invention demonstrated high safety, as shown in the MTS assay results targeting HepG2 liver cancer cell lines, that it did not exhibit cytotoxicity across various concentration ranges and showed toxicity so low that the CC50 value (50% cytotoxicity concentration) could not be calculated.

[0037] In addition, it was confirmed to effectively inhibit the proliferation of HBV in HepG2 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0038] In addition, it was confirmed to inhibit HBV DNA synthesis in Huh7 cells as well as HepG2 cells, indicating that it can be effective in various hepatocyte environments.

[0039] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0040] However, the effects obtainable from this invention are not limited to those described above, and other effects may exist.

[0041] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0042] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention.

[0043] Figure 3 is the result of a cytotoxicity test according to one experimental example of the present invention.

[0044] Figure 4 is the result of an experiment on the inhibition of HBV proliferation in a liver cancer cell line of Chemical Formula 1 according to one experimental example of the present invention.

[0045] Figure 5 is an experiment on the inhibition of HBV proliferation in a liver cancer cell line of Formula 2 according to one experimental example of the present invention.

[0046] Embodiments of the present invention are described below with reference to the attached drawings so that those skilled in the art can easily implement them. However, the present invention may be embodied in various different forms and is not limited to the embodiments described herein. Furthermore, in order to clearly explain the present invention in the drawings, parts unrelated to the explanation have been omitted, and similar parts throughout the specification are denoted by similar reference numerals.

[0047] Throughout this specification, when a part is described as being "connected" to another part, this includes not only cases where they are "directly connected," but also cases where they are "electrically connected" with other elements interposed between them.

[0048] Throughout the entire specification, when a component is described as being located "on," "on top," "on top," "under," "on bottom," or "on bottom" of another component, this includes not only cases where the component is in contact with the other component but also cases where another component exists between the two components.

[0049] Throughout this specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0050] As used herein, terms of degree such as “about,” “substantially,” etc., are used to mean at or near the stated value when inherent manufacturing and material tolerances are presented in the stated meaning, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which precise or absolute values ​​are mentioned to aid in understanding the invention. Furthermore, throughout this specification, “a step of” or “a step of” does not mean “a step for”.

[0051] Throughout this specification, the term “combination thereof” included in the Markush-type expression means one or more mixtures or combinations selected from the group consisting of the components described in the Markush-type expression, and means including one or more selected from the group consisting of said components.

[0052] Throughout the entire specification, the description "A and / or B" means "A, B, or A and B".

[0053] Hereinafter, the composition for inhibiting HBV proliferation of the present invention will be described in detail with reference to the embodiments, examples, and drawings. However, the present invention is not limited to these embodiments, examples, and drawings.

[0054]

[0055] As a technical means for achieving the above-mentioned technical problem, the first aspect of the present invention provides a composition for inhibiting HBV proliferation comprising a compound represented by the following chemical formula 1 as an active ingredient:

[0056] [Chemical Formula 1]

[0057]

[0058] (In the above chemical formula 1,

[0059] R is a substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0060] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl, halogen, or a combination thereof).

[0061] The composition for inhibiting HBV proliferation according to the present invention demonstrated high safety, as shown in the MTS assay results targeting HepG2 liver cancer cell lines, that it did not exhibit cytotoxicity across various concentration ranges and showed toxicity so low that the CC50 value (50% cytotoxicity concentration) could not be calculated.

[0062] In addition, it was confirmed to effectively inhibit the proliferation of HBV in HepG2 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0063] In addition, it was confirmed to inhibit HBV DNA synthesis in Huh7 cells as well as HepG2 cells, indicating that it can be effective in various hepatocyte environments.

[0064] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0065] According to one embodiment of the present invention, the compound represented by Formula 1 may include any one of the following compounds, but is not limited thereto:

[0066] ;

[0067] .

[0068] According to one embodiment of the present invention, HBc (Hepatitis B core) protein expression in HepG2 cells may be inhibited by a compound represented by the above chemical formula 1, but is not limited thereto.

[0069] The HBc protein is a core structural protein of the hepatitis B virus (HBV), essential for the formation of the viral capsid, and plays an important role in the replication and assembly processes of viral DNA.

[0070] HepG2 is a human liver cancer cell line and a widely used model system for HBV research. Because this cell line can express HBV proteins and produce viral particles, it is suitable for studying the effects of antiviral drugs.

[0071] The compound represented by Chemical Formula 1 according to the present invention can directly inhibit the expression of HBc protein or affect intracellular signaling pathways that regulate the expression of this protein, thereby inhibiting the formation of HBV capsid, which may make it difficult for the virus to replicate and assemble. This can consequently lead to a reduction in the production of new viral particles, which in turn leads to an effect of inhibiting the proliferation of HBV.

[0072] According to one embodiment of the present invention, the formation of a nucleocapsid in HepG2 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0073] The nucleocapsid is the core particle of HBV, having a structure in which the viral genome is packaged inside a capsid composed of HBc proteins. This is essential for the replication of viral DNA and the assembly of new viral particles.

[0074] Inhibition of nucleocapsid formation blocks important stages of the HBV life cycle, which can interfere with the packaging of the viral genome and the formation of mature viral particles.

[0075] Since this approach targets multiple stages of the viral life cycle simultaneously, effective antiviral activity can be expected, and it has the potential to reduce the likelihood of viral resistance developing.

[0076] According to one embodiment of the present invention, HBV DNA synthesis in HepG2 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0077] According to one embodiment of the present invention, HBV DNA synthesis in Huh7 cells may be inhibited by a compound represented by Chemical Formula 1, but is not limited thereto.

[0078] HBV DNA synthesis is a critical step in the replication process of the hepatitis B virus (HBV), and inhibiting this step can lead to effects such as suppressing viral replication, preventing the spread of infection, reducing liver damage, and improving long-term prognosis.

[0079] Meanwhile, HepG2 cells are a human liver cancer cell line and a widely used model system for HBV research; the inhibition of HBV DNA synthesis in HepG2 cells directly demonstrates the antiviral effect of the compound. This cell line can mimic various stages of the HBV life cycle, making it useful for studying the mechanism of action of the compound.

[0080] Huh7 cells are also a human liver cancer cell line and another important model system used in HBV research; the inhibition of HBV DNA synthesis in Huh7 cells indicates that the compound's effect is not limited to a specific cell line. This suggests that the compound's antiviral effect may be more general and broad.

[0081] The compound represented by Chemical Formula 1 according to the present invention can directly inhibit the synthesis of HBV intermediate DNA or inhibit the activity of HBV DNA polymerase, and can also indirectly inhibit DNA synthesis by interfering with HBV capsid formation.

[0082] The consistent effect in two different cell lines suggests that the compound may be effective in various hepatocyte environments, allowing for the expectation of broad effects despite individual differences among patients.

[0083]

[0084] In addition, the second aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to the first aspect of the present invention.

[0085] Regarding the pharmaceutical composition for preventing or treating HBV infection according to the second aspect of the present invention, detailed descriptions of parts that overlap with the first aspect of the present invention have been omitted, but even if such descriptions are omitted, the contents described in the first aspect of the present invention may be applied equally to the second aspect of the present invention.

[0086] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0087] According to one embodiment of the present invention, the carrier may comprise, but is not limited to, a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

[0088] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally include a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof, but is not limited thereto.

[0089] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may be administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof, but is not limited thereto.

[0090]

[0091] In addition, the third aspect of the present invention provides a health functional food for preventing or improving HBV infection, comprising a compound represented by Chemical Formula 1 of the first aspect of the present invention as an active ingredient.

[0092] Regarding the health functional food for preventing or improving HBV infection according to the third aspect of the present invention, detailed descriptions of parts that overlap with the first and / or second aspects of the present invention have been omitted, but even if such descriptions have been omitted, the contents described in the first and / or second aspects of the present invention may be applied equally to the third aspect of the present invention.

[0093] The present invention is to be explained in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0094]

[0095] [Example 1] ACT-C-001

[0096] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-phenylacrylamide (JYPf68)

[0097] Oxalyl chloride (83.6 μL, 0.974 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (100 mg, 0.325 mmol) dissolved in dichloromethane. After heating at reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of aniline (39.7 μL, 0.387 mmol) and DIPEA (119 μL, 0.682 mmol) at 0°C. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with a saturated NaHCO3 solution, a 1 M HCl solution, and brine, dried with Na2SO4, filtered, and concentrated to prepare a compound represented by the following structural formula. (10 mg, 8.1%)

[0098]

[0099] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0100] 1 H NMR (600 MHz, CDCl3) δ 8.17 (d,J = 2.8 Hz, 2H), 8.03 (s, 1H), 7.62 (d,J = 8.3 Hz, 2H), 7.48 (d,J = 3.4 Hz, 1H), 7.38-7.43 (m, 3H), 7.29 (td,J = 8.8, 3.0 Hz, 2H), 7.20 (t,J = 7.2 Hz, 1H)

[0101] [Example 2] ACT-C-009

[0102] (E)-2-cyano-N-cyclohexyl-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (JYPf88)

[0103] Oxalyl chloride (62.7 μL, 0.730 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (75 mg, 0.243 mmol) dissolved in dichloromethane. After heating at reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of cyclohexylamine (33.5 μL, 0.292 mmol) and DIPEA (89.0 μL, 0.511 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder compound represented by the following structural formula. (6.1 mg, 6.4%)

[0104]

[0105] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention.

[0106] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.1 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 3.4 Hz, 1H), 7.41 (d,J = 8.3 Hz, 1H), 7.27-7.28 (m, 1H), 7.20 (d,J = 3.4 Hz, 1H), 6.21 (d,J = 7.6 Hz, 1H), 3.90-3.92 (m, 1H), 1.98-2.01 (m, 2H), 1.77 (dt,J = 13.7, 3.7 Hz, 2H), 1.65 (td,J = 8.4, 4.1 Hz, 1H), 1.38-1.45 (m, 2H), 1.22-1.30 (m, 3H)

[0107] [Experimental Example 1] MTS assay: Cytotoxicity study

[0108] 2 x 10 per well in a 96-well microtiter plate 4 HepG2 cells were seeded with 100 μL of cell culture medium and cultured at 37°C under 5% CO2. The following day, compounds were treated at various concentrations (1, 5, 10, 50, 100 μM) and cultured for 48 hours. Subsequently, 10 μL of MTS reagent was added to each well, and the cells were cultured for 4 hours under standard cell culture conditions at 37°C. After briefly shaking the plates on a shaker, the absorbance was measured at a plate reader OD=490 nm.

[0109] Figure 3 is the result of a cytotoxicity study using the MTS assay according to one experimental example of the present invention.

[0110] Referring to FIG. 3, neither compound according to Examples 1 and 2 showed cytotoxicity in HepG2 liver cancer cell lines at 1, 5, 10, 50, and 100 μM, and CC 50 I couldn't calculate the value.

[0111] [Experimental Example 2]

[0112] Experiments were performed to confirm the HBV inhibitory efficacy of the compounds according to Examples 1 and 2 of the present invention.

[0113] After seeding HepG2 and Huh7 cells, 8 μg of 1.3mer HBV WT plasmid DNA was transfected into HepG2 cells and 3 μg into Huh7 cells the following day. pGEM4Z was used as a control. Simultaneously with transfection, HepG2 cells (2 x 10⁶ 6 / 6cm) Compound 1 at concentrations of 5μM and 10μM was applied to Huh7 cells (5x10 5In the / 6cm cell group, Compound 1 was treated at a concentration of 2.5 μM, and on the following day (24 hours later), when the medium was changed, Compound 1 was treated at the same concentration as before and cultured for an additional 48 hours. In Huh7 cells, AGK2 was treated at a concentration of 2.5 μM as a control and cultured in the same manner. After culturing for a total of 72 hours following transfection, cell lysates were prepared with 0.2% NP40-NTE buffer, followed by SDS-10% PAGE and Western blotting. Anti-acetylated α-tubulin, anti-α-tubulin, anti-sirtuin 2, anti-HBc, and anti-GAPDH antibodies were used as primary antibodies for Western blotting, and enhanced chemiluminescence (ECL) was observed. After performing 1% native agarose gel electrophoresis (NAGE) on the lysate, Western blotting (immunoblotting) was performed using anti-HBc antibodies, and core particle formation was observed using ECL. To observe the synthesis of HBV replicative intermediate DNA (RI DNA) by Southern blotting, HBV DNA was extracted from the isolated core particles, subjected to 1% NAGE, and then transferred to a nylon membrane. 32 HBV DNA synthesis was observed by autoradiography after hybridization with a specific probe for the full length of P-labeled HBV. Among the RI DNAs, partially double-stranded relaxed circular DNA (RC DNA) and doubled-stranded linear DNA (DL DNA) were labeled RC and DL, respectively.

[0114] Figure 4 shows the experimental results of the HBV proliferation inhibitory efficacy of the compound according to Example 1 of the present invention. In Figure 4, the 1st to 5th panels of HepG2 and Huh7 cells represent Western blot results, the 6th panel represents the core particles after NAGE-immunoblotting, and the 7th panel of HepG2 cells represents the core particles after NAGE. 32 HBV nucleic acids within the particles were detected by autoradiography after hybridization with a specific probe for the full length of P-labeled HBV, and the final panel of HepG2 and Huh7 cells was Southern blotting to observe HBV RI DNA replication. HBV proliferation was demonstrated through HBc protein expression (4th panel), core particle formation (6th panel), and HBV RI DNA Southern blotting (last panel), and it was confirmed that Sirtuin 2 increased when HBV proliferated (lane 2) (3rd panel, lane 2). Furthermore, when pG2 cells were treated with the compound according to Example 1, an AGK2-derived variant of the SIRT2 inhibitor, at the indicated concentration, SIRT2 inhibition was not distinct (3rd panel, lanes 3, 4), but HBV proliferation was inhibited at a concentration of 10 μM (lane 4). In the case of Huh7 cells, AGK2 was used for comparison and the inhibition of HBV proliferation by AGK2 was confirmed (lane 4). When Huh7 cells were treated with the compound according to Example 1 at the indicated concentration (2.5 μM), it was confirmed that although SIRT2 inhibition was not distinct, HBV proliferation was inhibited (lane 3).

[0115] Through this, it was confirmed that the compound according to Example 1 of the present invention inhibits HBV proliferation, and in HepG2 cells, it inhibits HBc(C) protein expression and the formation of core particles, and inhibits HBV DNA synthesis, and in Huh7 cells, it was confirmed that it inhibits HBV DNA synthesis.

[0116] Figure 5 shows the experimental results of the HBV proliferation inhibitory efficacy of the compound according to Example 2 of the present invention. In Figure 5, the 1st to 5th panels of HepG2 and Huh7 cells are Western blot results, the 6th panel is NAGE-immunoblotting of core particles, and the last panel is Southern blotting to observe HBV RI DNA replication. HBV proliferation was demonstrated through HBc protein expression (4th panel), core particle formation (6th panel), and HBV RI DNA Southern blotting (last panel), and it was confirmed that Sirtuin 2 increases when HBV proliferates (lane 2) (3rd panels, lanes 2). In addition, when HepG2 cells were treated with the compound according to Example 2, which is an AGK2-derived variant and a SIRT2 inhibitor, at the indicated concentrations, it was confirmed that SIRT2 was inhibited (3rd panels, lanes 3, 4) and HBV proliferation was inhibited at a concentration of 10 μM (lane 4). In the case of Huh7 cells, when Huh7 cells were treated with the compound according to Example 2 at the indicated concentrations (2.5 μM, 5 μM, 10 μM), it was confirmed that SIRT2 inhibition was distinct and HBV proliferation was inhibited (lanes 3-5).

[0117] Through this, it was confirmed that the compound according to Example 2 of the present invention retained its SIRT2 enzyme inhibitory function and inhibited HBV proliferation in HepG2 cells and Huh7 cells. In addition, it was confirmed that it inhibited HBc(C) protein expression and the formation of core particles, and inhibited HBV DNA synthesis.

[0118]

[0119] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical concept or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0120] The scope of the present invention is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and the concept of equivalents thereof should be interpreted as being included within the scope of the present invention.

Claims

1. A composition for inhibiting HBV proliferation comprising a compound represented by the following chemical formula 1 as an active ingredient: [Chemical Formula 1] (In the above chemical formula 1, R is a substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi, The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl, halogen, or a combination thereof).

2. In Paragraph 1, A composition for inhibiting HBV proliferation, wherein the compound represented by the above chemical formula 1 comprises any one of the following compounds: ; .

3. In Paragraph 1, The inhibition of HBc (Hepatitis B core) protein expression in HepG2 cells by a compound represented by the above chemical formula 1, Composition for inhibiting HBV proliferation.

4. In Paragraph 1, The formation of nucleocapsid in HepG2 cells is inhibited by a compound represented by the above chemical formula 1, Composition for inhibiting HBV proliferation.

5. In Paragraph 1, HBV DNA synthesis in HepG2 cells is inhibited by a compound represented by the above chemical formula 1, Composition for inhibiting HBV proliferation.

6. In Paragraph 1, HBV DNA synthesis in Huh7 cells is inhibited by a compound represented by the above chemical formula 1, Composition for inhibiting HBV proliferation.

7. A pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to any one of claims 1 to 6.

8. In Paragraph 7, The above pharmaceutical composition for the prevention or treatment of HBV infection further comprises a pharmaceutically acceptable carrier.

9. In Paragraph 8, A pharmaceutical composition for the prevention or treatment of HBV infection, wherein the carrier comprises a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

10. In Paragraph 7, The above pharmaceutical composition for preventing or treating HBV infection further comprises a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof.

11. In Paragraph 7, The above pharmaceutical composition for the prevention or treatment of HBV infection is administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof.

12. A health functional food for preventing or improving HBV infection, comprising a compound represented by Chemical Formula 1 of Claim 1 as an active ingredient.

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