KR102114394B1 - Benzophenone derivatives, preparation method thereof and pharmaceutical composition for treatment of hepatitis B virus comprising the same - Google Patents

Abstract

The present invention relates to a benzophenone derivative, a method for preparing the same, and a pharmaceutical composition for treating hepatitis B comprising the same, the compound represented by Formula 1 according to the present invention reduces HBsAg secretion and HBeAg secretion, and HBV virus level Since it has a reducing effect and has an excellent effect of inhibiting pgRNA production and cccDNA production, it may be useful as a pharmaceutical composition for preventing or treating HBV disease.

Description

Benzophenone derivatives, preparation method thereof and pharmaceutical composition for treating hepatitis B containing the same Benzophenone derivatives, preparation method thereof and pharmaceutical composition for treatment of hepatitis B virus comprising the same}

The present invention relates to a novel benzophenone derivative, a method for preparing the same, and a pharmaceutical composition for treating hepatitis B comprising the same.

Hepatitis B virus (HBV) is known as one of the most damaging viruses in the world with the highest infection rate since it was discovered by Blumberg in 1967 (now Nobel Prize for Medicine in 1976).

The genome of 3.2kb HBV is partial duplex DNA and has 4 open-reading frames (P, PreC / C, PreS1 / S2 / S, X). All four genes of HBV overlap, and the longest is the gene that expresses the polymerase of the virus. Since it is circular, all genes are expressed by frame-shift. Covalently closed circular DNA (cccDNA) is used as a template to produce four major RNAs: 3.5-, 2.4-, 2.1-, and 0.7-kb. These four RNAs are expressed by HBV-specific enhancer II / basal core, large surface antigen (L), small surface antigen (S), and enhancer I / X promoter, respectively.

HBV dissociates the capsid after infection and the gene moves into the nucleus, where the double strand viral DNA is converted into cccDNA in the host nucleus. It is known as a major cause of chronic infection because it is an episome-type minichromosome that not only makes all RNA of HBV but also cannot remove it with any current treatment.

The 3.5 kb pregenomic RNA makes polymerase and core protein (HBcAg), and the other 3.5 kb precore RNA makes e antigen (HBeAg). HBsAg is made from 2.4- and 2.1-kb RNA, and X protein (HBx) is made from 0.7-kb RNA. In addition to making proteins, pregenomic RNA plays another important role in life.

In other words, it acts as a template that is wrapped inside the capsid with polymerase and converted into a viral genome by reverse transcriptase. It is known that chronic infection with HBV is closely related to impaired liver function, hepatitis, cirrhosis and liver cancer.

The development of vaccines has made it possible to prevent some of hepatitis B, but once HBV has penetrated into hepatocytes, there is no way to completely remove HBV that has infiltrated the patient's hepatocytes. . In addition, when a hepatitis B patient receives a healthy liver, the recurrence rate of hepatitis B is very high because the remaining HBV is not completely removed and penetrates into the transplanted healthy liver cells.

When HBV penetrates into hepatocytes, it forms cccDNA (covalently closed circular DNA) in the cell nucleus. Once formed, cccDNA is used as a template for HBV genomic DNA replication, and continues to produce HBV until the life span of the hepatocyte is reached. do. Current treatments can inhibit HBV formation in the blood by inhibiting HBV genomic DNA replication, but there is no technique to remove cccDNA to prevent HBV replication at the source.

Korean Patent Publication No. 10-2017-0006299 Korean Patent Publication No. 10-2016-0127710

An object of the present invention is to provide a novel compound having hepatitis B treatment efficacy.

Another object of the present invention is to provide a method for preparing the compound.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating hepatitis B containing the compound as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving hepatitis B containing the compound as an active ingredient

In order to achieve the above object,

The present invention provides a compound represented by Formula 1 below, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

이고;R ¹ is

ego;

R ² is H, C _1-8 straight or branched alkyl, or aryl C _1-8 straight or branched alkyl).

In addition, the present invention, as shown in Scheme 1 below,

Dissolving compound 2 and compound 3 in a pyridine solvent and reacting at room temperature to obtain compound 1 (step 1);

It provides a method for producing a compound represented by the formula (1) comprising a.

[Scheme 1]

In Reaction Scheme 1,

R ¹ and R ² are as defined in Chemical Formula 1.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating hepatitis B comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

또는

이고;R ¹ is

or

ego;

R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl;

R ³ is C _1-5 straight chain or branched alkoxy, and the C _1-5 straight chain or branched alkoxy may be substituted with one or more hydroxy groups.

In addition, the present invention provides a health functional food composition for preventing or improving hepatitis B comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

The compound represented by Formula 1 according to the present invention reduces HBsAg secretion and HBeAg secretion, has an effect of decreasing HBV virus level, and has excellent effect of inhibiting pgRNA production and cccDNA production, thus preventing or treating HBV disease It may be useful as a pharmaceutical composition.

1 is a result of measuring the chemical structure and in vitro p38 MAPK enzyme inhibitory activity (% inhibition) of the compound according to the present invention.
Figure 2 (A) is a result of analyzing the secretion of HBsAg of the compound according to the present invention by ELISA.
Figure 2 (B) is a result showing the positive correlation between the inhibitory effect of p38 MAPK and HBsAg secretion of the compound according to the present invention.
Figure 3 (A) is a result showing the viability of HepG2 and HepG2.2.15 cells according to the treatment concentration (0, 0.2, 1, 5, 20 μM) of the compound NJK14047 according to the present invention.
Figure 3 (B) is a result of measuring the cytotoxicity of NJK14047 in HepG2.2.15, HepG2, AML-12, A549 and HELA cell lines.
Figure 3 (C) is a result of the inhibition of p38 MAPK activation over time and concentration of NJK14047 treatment of HepG2 cells transfected with HBV genome (pHBV-1.2x).
Figure 3 (D) is a graph showing the effect on the synthesis of mRNA of IL-6 according to the NJK14047 treatment concentration in HepG2.2.15 cells (* p <0.05, ** p <0.01).
Figure 3 (E) is a graph showing the effect on the synthesis of mRNA of IL-10 according to NJK14047 treatment concentration in HepG2.2.15 cells (* p <0.05, ** p <0.01).
Figure 4 (A) is a result of analyzing the HBsAg secretion in HepG2.2.15 cells according to the NJK14047 treatment concentration by ELISA (* p <0.05, ** p <0.01 and *** p <0.001).
Figure 4 (B) is the result of analyzing the HBsAg secretion in HepG2 cells transfected with pHBV-1.2x according to the NJK14047 treatment concentration by ELISA (* p <0.05, ** p <0.01 and *** p <0.001 ).
Figure 4 (C) is the result of analyzing the HBeAg secretion in HepG2 cells transfected with pHBV-1.2x according to the NJK14047 treatment concentration by ELISA (* p <0.05, ** p <0.01 and *** p <0.001 ).
Figure 4 (D) is a result of measuring the HBV virus level in HepG2.2.15 cells according to NJK14047 treatment concentration by qPCR (quantitative PCR) (* p <0.05, ** p <0.01 and *** p <0.001) .
Figure 4 (E) is a result of measuring the HBV virus level in HepG2 cells transfected with pHBV-1.2x according to NJK14047 treatment concentration by qPCR (quantitative PCR) (* p <0.05, ** p <0.01 and * ** p <0.001).
Figure 5 (A) is a result of measuring the pgRNA synthesis (3.5 kb) in HepG2.2.15 cells according to enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration by quantitative reverse transcription PCR (quantitative RT-PCR) (* p <0.05, ** p <0.01 and *** p <0.001, NS: Not significant).
FIG. 5 (B) is a quantitative reverse transcription PCR (quantitative RT-PCR) of pgRNA synthesis (3.5 kb) in HepG2 cells transfected with pHBV-1.2x according to enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration. The measured results (* p <0.05, ** p <0.01 and *** p <0.001).
Figure 5 (C) is a result of measuring the cccDNA synthesis (3.5kb) in HepG2.2.15 cells according to the enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration (quantitative PCR) (* p <0.05 , ** p <0.01 and *** p <0.001).
Figure 5 (D) is a result of measuring cccDNA synthesis (3.5kb) in HepG2 cells transfected with pHBV-1.2x according to enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration (* P <0.05, ** p <0.01 and *** p <0.001).
Figure 6 (A) is a stable expression of NTCP HepG2 in vitro HBV-infected model in enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration of extracellular virus DNA amount measured by quantitative PCR (quantitative PCR) and ELISA One result (* p <0.05, ** p <0.01).
FIG. 6 (B) shows the amount of secreted HBsAg according to enterkavir (ETV, 30 nM) treatment and NJK14047 treatment concentration in a HBV-infected model stably expressing NTCP in an HBV-infected model by quantitative PCR (quantitative PCR) and ELISA. One result (* p <0.05, ** p <0.01).

Hepatitis B virus (HBV) is a virus belonging to the Hepadnaviridae family and infiltrates hepatocytes and causes acute and chronic hepatitis.

The life cycle of HBV attaches to the surface of hepatocytes, penetrates into cells, infiltrates into cells, infiltrates into cell nuclei, reproduces DNA, proliferates into cells, assembles into cells, recombines into a complete virus form, and escapes out of hepatocytes It involves the process of excretion, and the process of repeatedly attaching to other hepatocytes and proliferating repeatedly.

HBV has a partially double-stranded DNA genome. HBV genomic DNA not only has gaps, but also has a covalent bond between a protein (oval shape) and RNA (wavy line) at both ends. It is also called Relaxed Circular DNA (RCDNA) because there is no torsional stress because it is connected to both ends.

In order for HBV to proliferate in hepatocytes, the DNA genome (RC DNA) of HBV must be injected into the cell nucleus of hepatocytes and from this, cvalent DNA (covalently closed circular DNA) must be generated. Once cccDNA is produced, the proliferation of HBV can continue until the cell has reached the end of its life. In order to produce cccDNA, HBV must first be penetrated into hepatocytes.

The envelope of HBV contains a Hepatitis B surface antigen (HBsAg), in order for HBV to penetrate into hepatocytes, HBsAg binds to heparan sulfate proteoglycan on the surface of hepatocytes and HBV The step of attaching to the surface of hepatocytes is essential. When HBV is attached to the surface of hepatocytes, HBV penetrates into the hepatocytes by binding the Pre-S1 domain of HBsAg to the sodium Taurocholate Cotransporting Polypeptide (NTCP), a receptor present on the surface of hepatocytes.

Covalently closed circular DNA (cccDNA) exists in the hepatocyte nucleus infected with HBV and acts as a template for HBV transcription. Inhibition of cccDNA formation in hepatitis B virus is a type of treatment that directly affects the life of the virus. It is distinct from immunotherapy such as interferon therapy, cytokine therapy, virus specific T cell immune recovery, and the like.

In addition, inhibition of cccDNA formation in the hepatitis B virus differs from the extinction method of the infected hepatitis B virus cccDNA. The cccDNA extinction method of the hepatitis B virus has a problem in that appropriate carrier development must be accompanied for clinical application.

When cccDNA formation of the hepatitis B virus is suppressed, HBV genomic DNA is not replicated and HBV reproduction becomes impossible, so HBV transmission to hepatocytes that are not yet infected does not occur any more. Hepatitis B virus cccDNA is degraded and disappears when cccDNA-formed hepatocytes have reached the end of their lifespan (about 5 months) and die.

Hereinafter, the present invention will be described in detail.

Compound or pharmaceutically acceptable salt thereof

The present invention provides a compound represented by Formula 1 below, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

이고;R ¹ is

ego;

R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl).

Preferably,

R ² is H, C _2-7 straight or branched chain alkyl, or phenyl C _1-3 straight or branched chain alkyl.

More preferably,

R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl.

Preferred examples of the compound represented by Formula 1 include the following compound groups, and their chemical structural formulas are shown in Table 1 below.

1) 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

2) N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

3) N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

4) 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

5) N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

6) N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

7) N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

8) 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The expression pharmaceutically acceptable salt is a concentration having a relatively non-toxic and harmless effect on a patient, and any side effect of the base compound of formula 1 does not degrade the beneficial effect of the base compound of formula 1 due to side effects caused by the salt. It means inorganic addition salt. As the free acid, inorganic and organic acids can be used. As the inorganic acid, hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, and phosphoric acid can be used. As the organic acid, citric acid, acetic acid, lactic acid, maleic acid, and fumarine can be used. Acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesul Folic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid can be used. In addition, these salts include alkali metal salts (sodium salt, potassium salt, etc.) and alkaline earth metal salts (calcium salt, magnesium salt, etc.). For example, acid addition salts include acetate, aspartate, benzate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Glucetate, Gluconate, Glucuronate, Hexafluorophosphate, Hybenzate, Hydrochloride / Chloride, Hydrobromide / Bromide, Hydroiodide / Iodine, Isethionate, Lactate, Malate, Malate , Malonate, mesylate, methyl sulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, famoate, phosphate / hydrogen phosphate / dihydrogen phosphate, saccharide , Stearate, succinate, tartrate, tosylate, trifluoroacete And aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salt, and the like. Among them, hydrochloride or trifluoroacetate is preferred.

In addition, the compound represented by Formula 1 of the present invention includes all salts, isomers, hydrates and solvates that can be prepared by conventional methods, as well as pharmaceutically acceptable salts.

The addition salt according to the present invention can be prepared by a conventional method, for example, dissolving the compound of Formula 1 in a water-miscible organic solvent, for example acetone, methanol, ethanol, or acetonitrile, adding an excess organic acid or inorganic acid It can be prepared by precipitation or crystallization after the addition of an aqueous acid solution. Subsequently, the solvent or excess acid may be evaporated from this mixture, followed by drying to obtain an added salt or a precipitated salt by suction filtration.

The present invention relates to any and all tautomers of compounds of Formula 1 having anti-HBV activity, and it can be understood that certain compounds of Formula 1 may exist in solvate and unsolvated forms, such as hydrated form. It can be understood that the present invention encompasses all these solvated forms with anti-HBV activity.

Manufacturing method

The present invention, as shown in Scheme 1 below,

Dissolving compound 2 and compound 3 in a pyridine solvent and reacting at room temperature to obtain compound 1 (step 1);

It provides a method for producing a compound represented by the formula (1) of claim 1 comprising a.

[Scheme 1]

(In Scheme 1 above,

R ¹ and R ² are as defined in Formula 1 of claim 1).

Pharmaceutical composition for prevention or treatment of hepatitis B disease

The present invention provides a pharmaceutical composition for the prevention or treatment of hepatitis B disease comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

또는

이고;R ¹ is

or

ego;

R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl;

R ³ is C _1-5 straight or branched alkoxy, and C _1-5 straight or branched alkoxy may be substituted with one or more hydroxy groups).

Preferably,

또는

이고;R ¹ is

or

ego;

R ² is H, C _2-7 straight or branched chain alkyl, or phenyl C _1-3 straight or branched chain alkyl;

R ³ is H, or C _1-4 straight or branched alkoxy, and C _1-4 straight or branched alkoxy may be substituted with 1 to 4 hydroxy groups.

More preferably,

또는

이고;R ¹ is

or

ego;

R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl;

R ³ is H, methoxy, or 2,3-dihydroxypropyl.

Preferred examples of the compound represented by Formula 1 include the following compound groups, and their chemical structural formulas are shown in Table 1 below.

1) 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

2) N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

3) N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

4) 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

5) N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

6) N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

7) N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

8) 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

9) N-cyclopropyl-4 '-(4-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide;

10) N-cyclopropyl-4 '-(3-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide;

11) N-cyclopropyl-4 '-(4- (2,3-dihydrooxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide.

The compound represented by the formula (1) according to the present invention has excellent p38 MAPK inhibitory ability to act as an inflammatory precursor (see Experimental Example 1), and confirmed the inhibitory effect of p38 MAPK on hepatocytes (see Experimental Example 4).

In addition, it was confirmed that the p38 MAPK and the HBsAg secretion had a positive correlation (see Experimental Example 2), and it was confirmed that the compound was able to inhibit HBsAg secretion through p38 MAPK inhibition.

It was confirmed that the antiviral activity of the compound NJK14047 according to the present invention was not caused by cytotoxicity (see Experimental Example 3). The compound NJK14047 according to the present invention reduces HBsAg secretion and HBeAg secretion in a dose-dependent manner (see Experimental Example 5, FIGS.

In addition, the HBV virus level was decreased according to the concentration of NJK14047 treatment, indicating that NJK14047 can inhibit not only viral antigen production but also viral replication and viral particle production (see Experimental Examples 6, 4D and 4E).

The compound NJK14047 according to the present invention inhibited pgRNA production (see Experimental Example 7), and showed an excellent effect of inhibiting cccDNA production (see Experimental Example 8).

Therefore, the compound represented by Formula 1 according to the present invention reduces HBsAg secretion and HBeAg secretion, has an effect of reducing HBV virus level, and has excellent effect of inhibiting pgRNA production and cccDNA production, thus preventing HBV disease Or it may be useful as a therapeutic pharmaceutical composition.

When the composition of the present invention is used as a pharmaceutical, the pharmaceutical composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient is administered in various oral or non-oral dosage forms as follows. It may be formulated and administered, but is not limited thereto.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules, liquids, suspensions, emulsifiers, syrups, granules, elixirs, etc., and these formulations are diluents (e.g. lactose, dext, in addition to active ingredients). Rose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols). Tablets may also contain a binder such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and optionally, such as starch, agar, alginic acid or its sodium salt. Disintegrants or boiling mixtures and / or absorbents, colorants, flavors, and sweeteners.

In addition, the dosage of the compound represented by Formula 1 according to the present invention to the human body may vary depending on the patient's age, weight, sex, dosage form, health status and disease degree, and is based on an adult patient with a body weight of 70 kg. When it is, it is generally 0.1 to 1,000 mg / day, preferably 1 to 500 mg / day, and may be dividedly administered once to several times a day at regular time intervals according to the judgment of a doctor or pharmacist.

Health functional food composition for prevention or improvement of hepatitis B disease

The present invention provides a health functional food composition for preventing or improving hepatitis B disease comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

또는

이고;R ¹ is

or

ego;

R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl;

R ³ is C _1-5 straight chain or branched alkoxy, and C _1-5 straight chain or branched alkoxy may be substituted with one or more hydroxy groups).

Preferably,

또는

이고;R ¹ is

or

ego;

R ² is H, C _2-7 straight or branched chain alkyl, or phenyl C _1-3 straight or branched chain alkyl;

R ³ is H, or C _1-4 straight or branched alkoxy, and C _1-4 straight or branched alkoxy may be substituted with 1 to 4 hydroxy groups.

More preferably,

또는

이고;R ¹ is

or

ego;

R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl;

R ³ is H, methoxy, or 2,3-dihydroxypropyl.

Preferred examples of the compound represented by Formula 1 include the following compound groups, and the chemical structural formulas thereof are shown in Table 1 below.

1) 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

2) N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

3) N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

4) 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

5) N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

6) N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

7) N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide

8) 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide

9) N-cyclopropyl-4 '-(4-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide;

10) N-cyclopropyl-4 '-(3-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide;

11) N-cyclopropyl-4 '-(4- (2,3-dihydrooxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide.

When the composition of the present invention is used as a dietary supplement, there are no particular restrictions on the type of food. Examples of foods to which the composition of the present invention can be added are drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various Soups, beverages, alcoholic beverages and vitamin complexes, dairy products and dairy products, etc., and include all functional foods in the normal sense.

The health functional food composition containing the compound represented by Formula 1 of the present invention may be added as it is to a food or used with other foods or food ingredients, and may be suitably used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (for prevention or improvement). In general, the amount of the composition in the dietary supplement can be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of maintaining health or for the purpose of controlling health, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

The health functional food composition containing the compound represented by the formula (1) of the present invention is an essential ingredient in the indicated proportions, and there is no particular limitation on other ingredients except that containing the composition of the present invention, and various flavors or natural carbohydrates, such as ordinary drinks And the like as additional components. Examples of the natural carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, etc .; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (such as taumatin and stevia extract (for example, rebaudioside A, glycyrrhizine)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 health functional food composition of the present invention.

In addition to the above, the health functional food composition containing the compound represented by Formula 1 of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate Etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain flesh for the preparation of natural fruit juice and fruit juice drinks and vegetable drinks.

These ingredients can be used independently or in combination. The proportion of these additives is not so critical, but is generally selected from 0.1 to about 20 parts by weight per 100 parts by weight of the health functional food composition containing the active substance of the present invention.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are only illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

<Example 1> 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1' biphenyl] -3-carboxamide (NJK13018)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.70 (d, 2H, J = 8.2 Hz), 7.65 (dd, 1H, J = 8.2, 1.8 Hz), 7.51 (s, 1H), 7.33-7.29 (m , 3H), 6.38 (s, 1H), 4.22 (t, 2H, J = 6.6 Hz), 2.89-2.88 (m, 1H), 2.28 (s, 3H), 2.27 (s, 3H), 1.77-1.64 ( m, 2H), 1.50-1.36 (m, 2H), 0.97 (t, 3H, J = 7.4 Hz), 0.85-0.84 (m, 2H), 0.61-0.60 (m, 2H)

<Example 2> N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13020)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.70 (d, 2H, J = 8.4 Hz), 7.65 (dd, 1H, J = 8.4, 1.8 Hz), 7.57 (d, 1H, J = 1.8 Hz), 7.31-7.29 (m, 3H), 6.36 (s, 1H), 4.25 (t, 2H, J = 6.8 Hz), 2.91-2.86 (m, 1H), 2.28 (s, 3H), 2.26 (s, 3H) , 1.81-1.72 (m, 1H), 1.64 (dt, 2H, J = 6.9, 6.6), 0.97 (d, 6H, J = 6.4), 0.88-0.82 (m, 2H), 0.61-0.60 (m, 2H )

<Example 3> N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13023)

¹ H-NMR (DMSO- d ₆ , 400 MHz) δ 11.19 (s, 1H), 8.34 (d, 1H, J = 4.1 Hz), 7.67 (d, 3H, J = 8.2 Hz), 7.63 (d, 1H , J = 1.3 Hz), 7.32 (t, 3H, J = 7.8 Hz), 2.78-2.77 (m, 1H), 2.21 (s, 3H), 2.13 (s, 3H), 0.62-0.59 (m, 2H) , 0.51-0.48 (m, 2H)

<Example 4> 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13031)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.68-7.62 (m, 2H), 7.57 (s, 2H), 7.40 (t, 1H, J = 7.6 Hz), 7.31-7.26 (m, 2H), 6.44 (s, 1H), 4.19 (t, 2H, J = 6.7 Hz), 2.90-2.86 (m, 1H), 2.28 (s, 3H), 2.25 (s, 3H), 1.74-1.67 (m, 2H), 1.48-1.39 (m, 2H), 0.96 (t, 3H, J = 7.4 Hz), 0.86-0.81 (m, 2H), 0.62-0.58 (m, 2H)

<Example 5> N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13032)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.66 (d, 1H, J = 7.9 Hz), 7.57 (d, 3H, J = 7.1 Hz), 7.40 (t, 1H, J = 7.6 Hz), 7.28 ( d, 2H, J = 8.2 Hz), 6.57 (s, 1H), 2.90-2.85 (m, 1H), 2.29 (s, 3H), 2.28 (s, 3H), 0.84-0.80 (m, 2H), 0.62 -0.58 (m, 2H)

<Example 6> N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13034)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.68-7.62 (m, 2H), 7.58-7.56 (m, 2H), 7.41 (t, 1H, J = 7.7 Hz), 7.32-7.26 (m, 2H) , 6.33 (s, 1H), 3.97 (d, 2H, J = 6.8 Hz), 2.92-2.86 (m, 1H), 2.28 (s, 3H), 2.26 (s, 3H), 2.09-2.02 (m, 1H) ), 0.96 (d, 6H, J = 6.7 Hz), 0.88-0.83 (m, 2H), 0.63-0.59 (m, 2H)

<Example 7> N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13035)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.68-7.62 (m, 2H), 7.57 (d, 2H, J = 2.0 Hz), 7.41 (t, 1H, J = 7.6 Hz), 7.32-7.26 (m , 2H), 6.38 (s, 1H), 4.23 (t, 2H, J = 6.8 Hz), 2.91-2.86 (m, 1H), 2.28 (s, 3H), 2.24 (s, 3H), 1.79-1.72 ( m, 1H), 1.64-1.59 (m, 2H), 0.95 (d, 6H, J = 6.6 Hz), 0.87-0.84 (m, 2H), 0.63-0.61 (m, 2H)

<Example 8> 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide (NJK13040)

¹ H-NMR (CDCl ₃ , 400 MHz) δ 7.68-7.65 (m, 2H), 7.58-7.56 (m, 2H), 7.43-7.35 (m, 5H,), 7.33-7.27 (m, 3H), 6.31 (s, 1H), 5.24 (s, 2H), 2.90-2.88 (m, 1H), 2.29 (s, 3H), 2.28 (s, 3H), 0.88-0.83 (m, 2H), 0.62-0.58 (m , 2H)

<Example 9> N-cyclopropyl-4 '-(4-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide (NJK14021)

A known compound N-cyclopropyl-4 '-(4-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide was prepared.

<Example 10> N-cyclopropyl-4 '-(3-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide (NJK14027)

A known compound N-cyclopropyl-4 '-(3-methoxybenzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide was prepared .

<Example 11> N-cyclopropyl-4 '-(4- (2,3-dihydrooxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide ( NJK14047)

A known compound N-cyclopropyl-4 '-(4- (2,3-dihydrooxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide was prepared. .

<Comparative Example 1> 4'-benzoyl-N-cyclopropyl-6-methyl- [1,1'-biphenyl] -3-carboxamide (NJK14018)

A known compound 4'-benzoyl-N-cyclopropyl-6-methyl- [1,1'-biphenyl] -3-carboxamide was prepared.

Chemical structures of Examples 1 to 11 and Comparative Example 1 Chemical structural formula Chemical structural formula Example 1
(NJK
13018)

Example 7
(NJK
13035)

Example 2
(NJK
13020)

Example 8
(NJK
13040)

Example 3
(NJK
13023)

Example 9
(NJK
14021)

Example 4
(NJK
13031)

Example 10
(NJK
14027)

Example 5
(NJK
13032)

Example 11
(NJK
14047)

Example 6
(NJK
13034)

Comparative Example 1
(NJK
14018)

<Experimental Example 1> p38 MAPK (mitogen-activated protein kinase) inhibitory evaluation

The compound prepared in Example 1-12 was evaluated for p38α MAPK inhibitory activity. Specifically, the p38α MAPK inhibitory ability of the compound prepared in Example 1-12 was obtained using “SelectScreen ™ Kinase Profiling Services (Life technologies)”, which is shown in Table 2 below (see FIG. 1).

compound % inhibition (1 μM) Example 1 (NJK13018) 21 Example 2 (NJK13020) 14 Example 3 (NJK13023) 42 Example 4 (NJK13031) 7 Example 5 (NJK13032) 25 Example 6 (NJK13034) 10 Example 7 (NJK13035) 6 Example 8 (NJK13040) 9 Example 9 (NJK14021) 84 Example 10 (NJK14027) 92 Example 11 (NJK14047) 97 Comparative Example 1 (NJK14018) 89

<Experimental Example 2> Confirmation of the correlation between p38 MAPK inhibitory ability and HBsAg (hepatitis B Surface Antigen) secretion

p38 MAPK inhibitory activity correlates with inhibition of HBsAg secretion. To investigate the anti-HBV activity of the compounds according to the present invention, HepG2.2.15 cells with HBV genotype D were cultured with the compounds for 48 hours.

All compounds except NJK13032 (Example 5) and NJK13040 (Example 8) were analyzed by HBsAg enzyme-linked immunosorbent assays (ELISA, Bio-kit). As shown in Fig. 2A, the compound according to the present invention inhibited the HBsAg secretion by more than 50% at a concentration of 10 μM. NJK14021 (Example 9) and NJK14027 (Example 10) showed about 50% inhibition at 2 μM.

In particular, NJK14047 (Example 11) showed the greatest inhibition of p38 MAPK and HBsAg secretion. As shown in Figure 2B, in vitro p38 MAPK enzyme inhibition and inhibition of HBsAg secretion by the compound showed a high positive correlation (r ² = 0.8547) so that the compound could inhibit HBsAg secretion through p38 MAPK inhibition. Was confirmed.

<Experiment 3> cytotoxicity confirmation

The cytotoxicity of the NJK14047 (Example 11) compound of the present invention to HepG2 and HepG2.2.15 cells was analyzed to exclude the possibility of antiviral activity caused by cytotoxicity.

Cytotoxicity was confirmed by using MTT (3- (4-5) -dimenthylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay to confirm the cytotoxicity of NJK14047 against various cells. Briefly, HepG2, HepG2.2.15, AML12, A549 or HeLa cells in a 96-well plate were cultured at different concentrations of NJK14047 for 48 hours.

HepG2, HepG2.2.15, A549 and HeLa cells were cultured in DMEM (Dulbecco's modified Eagle's 224 medium) supplemented with 10% fetal bovine serum (FBS) and penicillin / streptomycin (100 U / mL).

AML12 cells containing AML12 medium containing GlutaMAX, insulin (0.005 mg / mL), transferrin (0.005 mg / mL), selenium (5 ng / mL), dexamethasone (40 ng / mL) and 10% FBS (DMEM / F12; 1: 1).

hNTCP-HepG2-C4 cells (Dr, provided by Koichi Watashi) were cultured in AML12 medium containing G418 (500 μg / mL).

Then, the cell culture medium was replaced with fresh medium containing MTT (40 μg / mL) and incubated for another 4 hours. After aspiration, DMSO was added to lyse the cells and insoluble formazan. Absorbance at 570 nm was measured with a microplate reader.

NJK14047 compound did not show significant cytotoxicity to HepG2.2.15 cells at concentrations up to 20 μM. Therefore, 20 μM of NJK14047 was used as the maximum concentration in the following experimental example (see FIG. 3A).

In addition, the cytotoxic effect of NJK14047 on other cell lines was further analyzed. As shown in Figure 3B, the 50% cytotoxic concentration (CC ₅₀ ) value ranges from 53.7 to 110.4 μM, and the CC ₅₀ value of NJK14047 in normal mouse hepatocytes (AML-12 cells) is higher than other cell lines. Have (125.99 μM).

<Experiment 4> Confirmation of p38MAPK inhibition in hepatocytes

HepG2 cells transformed with a plasmid containing HBV genome (pHBV-112 1.2x, GenBank no. AY641558) to confirm p38 MAPK inhibition in hepatocytes were treated with 5 or 20 μM NJK14047 (Example 11) and subjected to immunoblotting. Were analyzed. NJK14047 treatment reduced p38 MAPK phosphorylation without affecting total protein levels, indicating that NJK14047 can inhibit p38 MAPK activation in HBV infected cells. (See Figure 3C).

In addition, NJK14047 treatment significantly inhibited the synthesis of mRNAs encoding interleukin (IL) -6 and interleukin (IL) -10 in HepG2.2.15 cells in a dose-dependent manner, confirming that NJK14047 can inhibit p38 MAPK-mediated inflammatory responses. did. (See Figures 3D and 3E).

<Experimental Example 5> HBsAg and HBeAg secretion measurement

In order to confirm the anti-HBV activity of the compound of the present invention NJK14047 (Example 11), HepG2.2.15 cells were treated by increasing the concentration of NJK14047, and the cell culture supernatant was recovered and the secretion of HBsAg was analyzed by ELISA.

As shown in Figure 4A, NJK14047 inhibited HBsAg secretion from HepG2.2.15 cells dose-dependently (IC ₅₀ = 5.3 μM, see Figure 4A). The effect of NJK14047 on HBeAg secretion using HepG2.2.15 cells could not be detected, which was also determined by ELISA (data not shown). The antiviral effect of NJK14047 was confirmed using an HBV genome transfected model with the genotype C viral genome.

After 24 hours of transfection, cells were treated with NJK14047 for 48 hours, and the supernatant was analyzed by ELISA. Unlike the results measured in HepG2.2.15 cells, compound NJK14047 showed a dose-dependent decrease in both HBsAg and HBeAg secretion (Figures 4B and 4C).

<Experimental Example 6> Measurement of virus particle production

The effect of NJK14047 (Example 11) on HBV virus production was analyzed to investigate the effect on HBV replication. HepG2 cells transfected with HepG2.2.15 247 and HBV genomes were treated with NJK14047 at the indicated concentrations for 48 hours. Virus particles in the collected culture supernatant were precipitated using PEG6000 as previously described (Wei Y, Tavis JE, Ganem D. 1996. Relationship between viral DNA synthesis and virion envelopment in hepatitis B viruses.J Virol 70: 6455 -6458.).

Virus pellets from HBV genome-transfected cells were treated with RQ1 DNase I (Promega, WI, USA) according to the manufacturer's instructions to remove residual DNA used for transfection. Viral DNA was quantified by qRT-PCR (quantitative real-time PCR) using the following primers: forward primer, 5'-TTAACAAGAATCCTCACAATACC-3 '; Reverse primer, 5'-GGAGGTTGGGGACTGCGAAT-3 '.

HBV virus levels were significantly reduced with NJK14047 treatment, indicating that NJK14047 can inhibit viral replication and viral particle production as well as viral antigen production (see Figure 4D). Similar results were derived from experiments with HepG2 cells transfected with pHBV-1.2x (see Figure 4E).

<Experimental Example 7> pgRNA production measurement

The effect of the compound of the present invention NJK14047 (Example 11) on HBV pgRNA production was analyzed. HBV pgRNA production in cells is slightly modified (Verrier ER, Colpitts CC, Bach C, Heydmann L, Weiss A, Renaud M, Durand SC, Habersetzer F, Durantel D, Abou-Jaoude G, Lopez Ledesma MM, Felmlee DJ, Soumillon M, Croonenborghs T, Pochet N, Nassal M, Schuster C, Brino L, Sureau C, Zeisel MB, Baumert TF. 2016.A targeted functional RNA interference screen uncovers glypican 5 as an entry factor for hepatitis B and D viruses.Hepatology 63 : 35-48.). Total RNA was isolated from HepG2.2.15 or HepG2 cells transfected with the HBV genome using TRIzol and treated with RQ1 DNaseI (Promega). RNA was purified by conventional phenol-chloroform isoamyl alcohol extraction and ethanol precipitation, and cDNA was synthesized with an RNA sample (1 μg).

The amount of pgRNA was measured by TaqMan real time RT-qPCR using the following primers and probes: forward primer, 5'-GGTCCCCTAGAAGAAGAACTCCCT-3 '; Reverse primer, 5'-CATTGAGATTCCCGAGATTGAGAT-3 '; TaqMan probe 5 '-[FAM] -TCTCAATCGCCGCGTCGCAGA- [TAMRA] -3'. All values were normalized to GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) as a control.

HepG2.2.15 cells (FIG. 5A) and pHBV-1.2x-transfected HepG2 cells (FIG. 5B) were treated with NJK14047 or Entecavir (ETV, 30nM) for 48 hours, resulting in 3.5 kb pgRNA in NJK14047 treated cells. A significant decrease in was observed compared to vehicle-treated cells. .

<Experimental Example 8> Quantification of HBV nuclear cccDNA

HepG2 cells transfected with HBV cccDNA of HepG2.2.15 or linearized HBV genome were partially modified and quantified (Belloni L, Allweiss L, Guerrieri F, Pediconi N, Volz T, Pollicino T, Petersen J, Raimondo G, Dandri M, Levrero M. 2012.IFN-alpha inhibits HBV transcription and replication in cell culture and in humanized mice by targeting the epigenetic regulation of the nuclear cccDNA minichromosome.J Clin Invest 122: 529-537.). The collected cells were washed with cold phosphate buffered saline and lysed by incubating in 270 μl of lysis buffer (50 mM Tris-HCl, pH 7.4, 1 mM ethylene diamine tetraacetic acid [EDTA], 1% NP-40) for 10 min on ice. Ordered.

After brief centrifugation (10000 × g, 1 min), the pelletized nucleus was degraded Buffer B (10 mM Tris-HCl, 10 mM EDTA, 150 mM NaCl, 0.5% sodium dodecyl sulfate, and 0.5 mg / mL protein K) And re-suspended at 37 ° C overnight.

DNA extracted by phenol-chloroform extraction and ethanol extraction was treated with 10 U Plasmid-Safe ATP-dependent DNase I (10U; Epicentre, WI, USA) for 45 minutes, and then DNase was inactivated at 70 ° C.

cccDNA was purified using MEGA Quick Spin Kit (Intron) and quantified by real-time quantitative PCR using the following primers and probes: cccF1, 5'-CCGTCTGTGCCTTCTCAT-3 '; cccR1, 5'-CACAGCTTGGAGGCTTGAAC-3 '; cccProbe, 5'-FAM280 CGTGTGCACTTCGCTTCACCTCTGC-TAMRA-3 '.

5C and 5D, it was found that treatment with NJK14047 significantly reduced cccDNA in both experiments.

In addition, treatment with 5 μM NJK14047 reduced cccDNA by 70% and 90% in HepG2.2.15 cells and pHBV-1.2x-transfected cells, respectively. Therefore, it was confirmed by the above results that p38MAPK inhibition by the compound NJK14047 of the present invention can inhibit cccDNA production and pgRNA synthesis.

<Experimental Example 9> In vitro HBV infection analysis

Culture supernatants were collected every 2 days for 14 days from HepG2.2.15 cells (4 x 107 cells / 150-mm dish). The supernatant was simply centrifuged and filtered to remove cell debris (0.45 μm).

Virus particles were concentrated by ultracentrifugation (100,000 × g, 3 hours, 4 ° C.). Virus particles were resuspended in AML12 medium containing DMSO (2%) and PEG8000 (4%). NTCP-HepG2-C4 cells in AML12 medium containing DMSO (2%) were infected with HBV (6 × 103 genome equivalents / cell) and centrifuged (1300 rpm / 1 hour).

After incubation for 24 hours, cells were washed and maintained for 7 days in AML12 medium containing NJK14047. Extracellular viral DNA and HBsAg levels were determined by qPCR (quantitative PCR) and ELISA as described above.

As a result, NJK14047 treatment confirmed a significant decrease in both HBV virion particle production and HBsAg secretion in a dose-dependent manner (see FIGS. 6A and B).

Therefore, the compound represented by Formula 1 according to the present invention reduces HBsAg secretion and HBeAg secretion, has an effect of reducing HBV virus level, and has excellent effect of inhibiting pgRNA production and cccDNA production, thus preventing HBV disease Or it may be useful as a therapeutic pharmaceutical composition.

Meanwhile, the compound represented by Chemical Formula 1 according to the present invention may be formulated in various forms according to the purpose. The following are examples of some formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, and the present invention is not limited thereto.

<Production Example 1> Preparation of powder

2 g of compound of formula 1

1 g of lactose

A powder was prepared by mixing the above components and filling the gas-tight fabric.

<Formulation Example 2> Preparation of tablets

100 mg of compound of formula 1

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2mg

After mixing the above components, tablets were prepared by tableting according to a conventional tablet manufacturing method.

<Formulation Example 3> Preparation of capsules

100 mg of compound of formula 1

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2mg

After mixing the above components, the capsules were prepared by filling the gelatin capsules according to a conventional capsule preparation method.

<Formulation Example 4> Preparation of injection

100 mg of compound of formula 1

Mannitol 180 mg

Na2HPO4 ㆍ 2H2O 26 mg

Distilled water 2974 mg

According to the conventional method of preparing an injection, the above ingredients were contained in the indicated amounts to prepare an injection.

Example of manufacturing health functional food

The compound represented by the formula (1) according to the present invention can be prepared as a health functional food in various forms depending on the purpose. The following is an example of a method for manufacturing some health functional foods containing the active substance according to the present invention as an active ingredient, and the present invention is not limited thereto.

<Production Example of Health Functional Food> Preparation of Health Functional Food

100 mg of compound of formula 1

Vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5 mg

Suitable amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the vitamin and mineral mixture is a composition suitable for a relatively healthy functional food in a preferred embodiment, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for manufacturing a healthy functional food. Then, the granules can be prepared and used in the production of a health functional food composition according to conventional methods.

<Production Example of Health Functional Food> Preparation of Health Functional Drink

100 mg of compound of formula 1

100 mg citric acid

Oligosaccharide 100 mg

Plum concentrate 2 mg

Taurine 100 mg

Purified water was added to make 500 mL

After mixing the above components according to a conventional health drink manufacturing method, after stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered, obtained in a sterilized 1 container, sealed and sterilized, and then refrigerated and stored in the present invention. It is used in the manufacture of health drink compositions. Although the above composition ratio is a mixture of components suitable for a comparatively preferred beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, country of demand, and usage.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims, not the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Claims (10)

A compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In the above formula 1,
R ¹ is

ego;
R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl).
According to claim 1,
R ² is H, C _2-7 straight chain or branched chain alkyl, or phenyl C _1-3 straight chain or branched chain alkyl, or a pharmaceutically acceptable salt thereof.
According to claim 1,
R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl, or a pharmaceutically acceptable salt thereof.
According to claim 1,
The compound represented by Formula 1 is a compound or a pharmaceutically acceptable salt thereof, characterized in that any one selected from the following group of compounds:
1) 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide;
2) N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
3) N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
4) 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide;
5) N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
6) N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
7) N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide; And
8) 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide.
As shown in Scheme 1 below,
Dissolving compound 2 and compound 3 in a pyridine solvent and reacting at room temperature to obtain compound 1 (step 1);
Method for preparing a compound represented by Formula 1 of claim 1 comprising:
[Scheme 1]

(In Scheme 1 above,
R ¹ and R ² are as defined in Formula 1 of claim 1).
A pharmaceutical composition for preventing or treating hepatitis B comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In the above formula 1,
R ¹ is

ego;
R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl).
The method of claim 6,
R ² is H, C _2-7 straight chain or branched chain alkyl, or phenyl C _1-3 straight chain or branched chain alkyl.
The method of claim 6,
R ² is H, C _4-5 linear or branched alkyl, or phenylmethyl.
The method of claim 6,
The compound represented by Formula 1 is any one selected from the following group of compounds:
1) 4 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide;
2) N-cyclopropyl-4 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
3) N-cyclopropyl-4 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
4) 3 '-(1- (butoxyimino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide;
5) N-cyclopropyl-3 '-(1- (hydroxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
6) N-cyclopropyl-3 '-(1-((isobutoxyimino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide;
7) N-cyclopropyl-3 '-(1-((isopentyloxy) imino) ethyl) -6-methyl- [1,1'biphenyl] -3-carboxamide; And
8) 3 '-(1-((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl] -3-carboxamide.
Hepatitis B prevention or improvement health functional food composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In the above formula 1,
R ¹ is

ego;
R ² is H, C _1-8 straight or branched chain alkyl, or aryl C _1-8 straight or branched chain alkyl).

Images