KR20190023760A - 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 manufacturing method thereof, and a pharmaceutical composition for treating hepatitis B comprising the same. A compound represented by chemical formula 1 according to the present invention reduces HBsAg secretion and HBeAg secretion, is excellent in an effect of inhibiting pgRNA production and cccDNA production, thereby being useful as the pharmaceutical composition for preventing or treating HBV disease.

Description

[0001] The present invention relates to a benzophenone derivative, a process for preparing the same, and a pharmaceutical composition for treating hepatitis B comprising the same,

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

Hepatitis B virus (HBV) is known to be one of the viruses that has been causing harm to humans with the highest infection rates in the world since it was discovered by Blumberg (received the Nobel Prize for Medicine in 1976) in 1967.

The 3.2 kb HBV genome is a partial duplex DNA and has four open reading frames (P, PreC / C, PreS1 / S2 / S, X). The four genes of HBV are all overlapping, and the longest one is the gene that expresses the polymerase of the virus. Because they are circular, all genes are differentially 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.

After infection, the capsid is disassembled and the gene is transferred into the nucleus, and the double strand virus DNA is converted into cccDNA in the nucleus of the host. This is an episomal minichromosome that is known to be the major cause of chronic infection as it not only makes all the RNA of HBV, but it can not destroy it with any current therapeutic agent.

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

In other words, it acts as a template that is encapsulated in the capsid with the polymerase and converted into the viral genome by reverse transcriptase. Chronic infection by HBV is known to be closely related to liver function impairment, hepatitis, liver cirrhosis and liver cancer.

However, once the HBV has penetrated into the hepatocytes, there is no way to completely remove the HBV that has penetrated into the hepatocytes of the patient. Thus, the fundamental cure for hepatitis B is very difficult at present . In addition, when hepatitis B patients are transplanted with healthy liver, the recurrence rate of hepatitis B is very high, because the remaining HBV infiltrates into the transplanted healthy hepatocytes.

When HBV infiltrates into hepatocytes, cccDNA (covalently closed circular DNA) is formed in the nucleus. Once formed, cccDNA is used as a template for HBV genomic DNA replication to produce HBV continuously until the lifespan of the hepatocytes do. Current therapies include the inhibition of HBV genomic DNA replication, which can inhibit HBV formation in the blood, but there is no technique to remove cccDNA to prevent replication of HBV.

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

It is an object of the present invention to provide novel compounds having hepatitis B treatment efficacy.

Another object of the present invention is to provide a process for producing the above compound.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating hepatitis B comprising the compound as an active ingredient.

It is still another object of the present invention to provide a health functional food composition for preventing or ameliorating hepatitis B comprising the compound as an active ingredient

In order to achieve the above object,

The present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(In the 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.

Also, as shown in the following Reaction Scheme 1,

A step of dissolving Compound 2 and Compound 3 in a pyridine solvent and reacting at room temperature to obtain Compound 1 (Step 1);

Wherein R < 1 > and R < 2 >

[Reaction Scheme 1]

In the above Reaction Scheme 1,

Wherein R ¹ and R ² are the same as defined in the above formula (1).

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating hepatitis B comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In 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.

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

The compound of formula (I) according to the present invention reduces the HBsAg secretion and HBeAg secretion, reduces the level of HBV virus, and has an excellent effect of inhibiting pgRNA production and inhibiting cccDNA production. Therefore, May be useful as pharmaceutical compositions.

Figure 1 shows the chemical structure of the compounds according to the present invention and the results of measuring in vitro p38 MAPK enzyme inhibitory activity (% inhibition).
FIG. 2 (A) shows the results of analysis of the secretion of HBsAg of the compound according to the present invention by ELISA.
FIG. 2 (B) shows the positive correlation between the p38 MAPK inhibitory activity and the HBsAg secretion inhibitory effect of the compound of the present invention.
FIG. 3 (A) shows the viability of HepG2 and HepG2.2.15 cells according to the treatment concentration (0, 0.2, 1, 5, 20 μM) of compound NJK14047 according to the present invention.
FIG. 3 (B) shows the results of measuring the cytotoxicity of NJK14047 in HepG2.2.15, HepG2, AML-12, A549 and HELA cell lines.
Fig. 3 (C) shows the results of inhibition of p38 MAPK activation by HepG2 cells transfected with the HBV genome (pHBV-1.2x) with NJK14047 treatment concentration and time.
FIG. 3 (D) is a graph showing the effect of IL-6 mRNA synthesis according to the concentration of NJK14047 treatment in HepG2.2.15 cells (* p <0.05, ** p <0.01).
FIG. 3 (E) is a graph showing the effect of IL-10 mRNA synthesis according to the treatment concentration of NJK14047 in HepG2.2.15 cells (* p <0.05, ** p <0.01).
FIG. 4 (A) is the result of ELISA analysis of HBsAg secretion in HepG2.2.15 cells according to the treatment concentration of NJK14047 (* p <0.05, ** p <0.01 and *** p <0.001).
FIG. 4 (B) is the result of ELISA analysis of HBsAg secretion in HepG2 cells transfected with pHBV-1.2x according to the treatment concentration of NJK14047 (* p <0.05, ** p <0.01 and *** p <0.001 ).
Figure 4 (C) shows the results of ELISA analysis of HBeAg secretion in HepG2 cells transfected with pHBV-1.2x according to the treatment concentration of NJK14047 (* p <0.05, ** p <0.01 and *** p <0.001 ).
Figure 4 (D) shows the results of qPCR (* p <0.05, ** p <0.01 and *** p <0.001) of HBV virus levels in HepG2.2.15 cells according to the treatment concentration of NJK14047 .
4 (E) shows the results of qPCR (quantitative PCR) of HBV levels in hepG2 cells transfected with pHBV-1.2x according to the treatment concentration of NJK14047 (* p <0.05, ** p <0.01 and * ** p < 0.001).
FIG. 5A is a result of quantitative RT-PCR measurement of pGRNA synthesis (3.5 kb) in HepG2.2.15 cells according to treatment with enterivir (ETV, 30 nM) and NJK14047 treatment concentration (* p <0.05, ** p <0.01 and *** p <0.001, NS: Not significant).
Fig. 5 (B) shows quantitative RT-PCR of pvRNA synthesis (3.5 kb) in HepG2 cells transfected with EnterBir (ETV, 30 nM) and pHBV-1.2x according to the treatment concentration of NJK14047 (* P <0.05, ** p <0.01 and *** p <0.001).
5C shows the result of quantitative PCR (* p < 0.05) for the treatment of enterococci (ETV, 30 nM) and ccc DNA synthesis (3.5 kb) in HepG2.2.15 cells according to the treatment concentration of NJK14047 , ** p < 0.01 and *** p < 0.001).
Figure 5 (D) shows the results of quantitative PCR of ccc DNA synthesis (3.5 kb) in HepG2 cells transfected with EnterBir (ETV, 30 nM) and NBK14047 treatment concentration and pHBV-1.2x (* P < 0.05, ** p < 0.01 and *** p < 0.001).
Figure 6 (A) shows the quantification of extracellular viral DNA by treatment with enterivir (ETV, 30 nM) and NJK14047 treatment in a HepG2 in vitro HBV-infected model stably expressing NTCP by quantitative PCR and ELISA (* P <0.05, ** p <0.01).
FIG. 6 (B) shows quantitative PCR and ELISA for the amount of HBsAg secreted by enterovir (ETV, 30 nM) treatment and NJK14047 treatment in a HepG2 in vitro HBV-infected model stably expressing NTCP (* P <0.05, ** p <0.01).

Hepatitis B virus (HBV) is a virus belonging to the family of Hepadnaviridae that infiltrate hepatocytes and cause acute and chronic hepatitis.

The life history of HBV is a process of infiltration into the cell that sticks to the surface of the hepatocyte, the process of peeling the cell in the cytoplasm and entering into the nucleus to replicate DNA, the assembly process of stacking the shell in the cytoplasm and recombining into a complete virus form, The process of excretion, and the process of repeated proliferation, stuck to another hepatocyte again.

HBV has a partially double-stranded DNA genome. HBV genomic DNA has not only a gap but also an oval shape or a wavy line at both ends. And is called Relaxed Circular DNA (RCDNA) because there is no torsional stress because both ends are not connected.

For HBV proliferation in hepatocytes, the DNA genome of HBV (RC DNA) must be injected into the nucleus of the hepatocyte, from which cccalently closed circular DNA (cccDNA) should be generated. Once cccDNA is generated, the proliferation of HBV can continue until the cell's lifespan is reached. In order to generate cccDNA, HBV must first penetrate hepatocytes.

The envelope of HBV contains a surface antigen (HBsAg). In order for HBV to penetrate hepatocytes, HBsAg binds to heparan sulfate proteoglycan on the surface of hepatocytes and HBV Is attached to the surface of the hepatocyte. When HBV is attached to the surface of hepatocytes, HBV binds to the hepatocytes by binding to the sodium Taurocholate Cotransporting Polypeptide (NTCP), a receptor on the surface of the hepatocytes.

Covalently closed circular DNA (cccDNA) is present in hepatocyte nuclei infected with HBV and acts as a template for HBV transcription. Inhibition of cccDNA formation by hepatitis B virus is a type of therapy that directly affects the life cycle of the virus. This 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 by hepatitis B virus differs from that of the infected hepatitis B virus cccDNA. The cccDNA extinction method of hepatitis B virus has to be accompanied by development of suitable carrier for clinical application.

If cccDNA formation of hepatitis B virus is suppressed, replication of HBV genomic DNA is not achieved and HBV reproduction becomes impossible, so that HBV propagation to uninfected hepatocytes is no longer occurred. When hepatocytes already formed with cccDNA are dead due to their life (about 5 months), cccDNA of hepatitis B virus is also decomposed and disappeared.

Hereinafter, the present invention will be described in detail.

Compound or a pharmaceutically acceptable salt thereof.

The present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(In the 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 chain or branched chain alkyl.

More preferably,

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

Preferable examples of the compound represented by the above formula (1) include the following compounds, and their chemical structures 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) Synthesis of 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) Synthesis of 3 '- (1 - ((benzyloxy) imino) ethyl) -N- cyclopropyl-6-methyl- [1,1'biphenyl]

The compound represented by the 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 term pharmaceutically acceptable salt means a concentration that is relatively non-toxic to a patient and has a beneficial effect that is harmless to the patient, such that the side effects caused by the salt are any organic or inorganic salt of the base compound of Formula 1 that does not impair the beneficial effects of the base compound of Formula An inorganic addition salt. Examples of the inorganic acid include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid and phosphoric acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, tartaric acid, succinic acid, malonic acid, succinic acid, malonic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid. These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / , Maleate, maleate, methylsulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, , Stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Hydrochloride or trifluoroacetate is preferred.

In addition, the compound represented by Formula 1 of the present invention includes not only pharmaceutically acceptable salts, but also all salts, isomers, hydrates and solvates which can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of Chemical Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile, etc., Followed by precipitation or crystallization. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

It is understood that the present invention relates to any and all arc variants of the compounds of formula (I) having anti-HBV activity and that certain compounds of formula (I) may exist in solvated form and in unsolvated form, e.g., hydrated form. The present invention may be understood to encompass all such solvated forms with anti-HBV activity.

Manufacturing method

As shown in the following Reaction Scheme 1,

A step of dissolving Compound 2 and Compound 3 in a pyridine solvent and reacting at room temperature to obtain Compound 1 (Step 1);

(1), wherein R &lt; 1 &gt;

[Reaction Scheme 1]

(In the above Reaction Scheme 1,

Wherein R &lt; ¹ &gt; and R &lt; ² &gt; are the same as defined in formula (1).

A pharmaceutical composition for the prevention or treatment of hepatitis B disease

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

[Chemical Formula 1]

(In the 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 &lt; ³ &gt; is _C1-5 straight chain or branched alkoxy, and the _C1-5 linear or branched alkoxy may be substituted with at least one hydroxy group.

Preferably,

또는

이고;R ¹ is

or

ego;

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

R ³ is H, or C _1-4 straight chain or branched alkoxy, and the C _1-4 straight chain 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.

Preferable examples of the compound represented by the above formula (1) include the following compounds, and their chemical structures 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) Synthesis of 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) Synthesis of 3 '- (1 - ((benzyloxy) imino) ethyl) -N- cyclopropyl-6-methyl- [1,1'biphenyl]

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-dihydroxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide.

The compound represented by Formula 1 according to the present invention has excellent p38 MAPK inhibitory ability as a precursor of inflammation (see Experimental Example 1), and confirmed the effect of inhibiting p38 MAPK in hepatocytes (see Experimental Example 4).

In addition, it was confirmed that p38 MAPK and HBsAg secretion were positively correlated (see Experimental Example 2), and it was confirmed that the compound could 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 dose-dependently decreases HBsAg secretion and HBeAg secretion (see Experimental Example 5, Fig. 4B and Fig. 4C).

In addition, the level of HBV virus was decreased according to the treatment concentration of NJK14047, indicating that NJK14047 can inhibit viral replication and viral particle production as well as viral antigen production. Experimental Example 6, Fig. 4D and Fig. 4E.

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

Therefore, the compound of formula (I) according to the present invention reduces the secretion of HBsAg and the secretion of HBeAg, reduces the level of HBV virus, inhibits pgRNA production and inhibits cccDNA production, Or pharmaceutical compositions for therapeutic use.

When the composition of the present invention is used as a medicine, the pharmaceutical composition containing the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient may be administered orally or non-orally May be formulated and administered, but the present invention is not limited thereto.

Examples of the formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules and elixirs. These formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, (For example, silica, talc, stearic acid and magnesium or calcium salts thereof and / or polyethylene glycol), and the like. The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain additives such as starch, agar, alginic acid or its sodium salt A disintegrating or boiling mixture and / or an absorbent, a colorant, a flavoring agent, and a sweetening agent.

The dose of the compound of formula (I) according to the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease severity. , It is generally 0.1 to 1,000 mg / day, preferably 1 to 500 mg / day, and may be administered once or several times a day at a predetermined time interval according to the judgment of a doctor or pharmacist.

A health functional food composition for preventing or ameliorating hepatitis B disease

The present invention provides a health functional food composition for preventing or ameliorating a hepatitis B disease comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(In the 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 &lt; ³ &gt; is _C1-5 straight chain or branched alkoxy, and the _C1-5 linear or branched alkoxy may be substituted with at least one hydroxy group.

Preferably,

또는

이고;R ¹ is

or

ego;

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

R ³ is H, or C _1-4 straight chain or branched alkoxy, and the C _1-4 straight chain 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.

Preferable examples of the compound represented by the above formula (1) include the following compounds, and their chemical structures 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) Synthesis of 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) Synthesis of 3 '- (1 - ((benzyloxy) imino) ethyl) -N- cyclopropyl-6-methyl- [1,1'biphenyl]

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-dihydroxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide.

When the composition of the present invention is used as a health functional food, the kind of the food is not particularly limited. Examples of foods to which the composition of the present invention can be added include food products such as drinks, meat, sausages, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, Soups, beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, all of which include health functional foods in a conventional sense.

The health functional food composition containing the compound represented by the general formula (1) of the present invention can be added as it is to the food or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement). Generally, the amount of the composition in the health functional food may be 0.1 to 90 parts by weight of the total food. However, in the case of long-term intake for the purpose of health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional food composition containing the compound represented by formula (1) of the present invention is not particularly limited as long as it contains the composition of the present invention as an essential ingredient in the indicated ratio, and may be various flavors or natural carbohydrates Or the like as an additional component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above . The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 of the health functional food composition of the present invention.

In addition to the above, the health functional food composition containing the compound represented by the formula (1) of the present invention can be used as a nutritional supplement, a vitamin, a mineral (electrolyte), a flavor such as a synthetic flavor and a natural flavor, Etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the health functional food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

These components may be used independently or in combination. The proportion of such additives is not so important, but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the health functional food composition containing the effective substance of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example 1 Synthesis of 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

Example 2 Synthesis of N-cyclopropyl-4 '- (1 - ((isopentyloxy) imino) ethyl) -6-

_{^{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 )

Example 3 Synthesis of 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 Synthesis of 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), (M, 2H), 0.96 (t, 3H, J = 7.4 Hz), 0.86-0.81 (m, 2H), 0.62-0.58

Example 5 Synthesis of 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 ( 2H, J = 8.2 Hz), 6.57 (s, 1H), 2.90-2.85 (m, 1H), 2.29 (s, 3H), 2.28 -0.58 (m, 2H)

Example 6 Synthesis of 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) 3H), 2.26 (s, 3H), 2.09-2.02 (m, IH), 6.33 (s, IH), 3.97 (d, 2H, J = 6.8 Hz), 2.92-2.86 ), 0.96 (d, 6H, J = 6.7Hz), 0.88-0.83 (m, 2H), 0.63-0.59

Example 7 Synthesis of 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, IH), 4.23 (t, 2H, J = 6.8 Hz), 2.91-2.86 (m, IH), 2.28 (s, 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 Synthesis of 3'- (1 - ((benzyloxy) imino) ethyl) -N-cyclopropyl-6-methyl- [1,1'biphenyl]

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

Example 9 Synthesis of 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 Synthesis of 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 Synthesis of N-cyclopropyl-4 '- (4- (2,3-dihydroxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide NJK14047)

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

Comparative Example 1 Synthesis of 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.

In 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 Evaluation of p38 MAPK (mitogen-activated protein kinase)

The p38? MAPK inhibiting ability of the compound prepared in Example 1-12 was evaluated. Specifically, the p38? MAPK inhibitory effect of the compound prepared in Example 1-12 was determined using SelectScreen ™ Kinase Profiling Services (Life technologies), and the results are shown in Table 2 below (see FIG. 1).

compound % inhibition (1 [mu] M) 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 Correlation between p38 MAPK Inhibition and HBsAg (Hepatitis B Surface Antigen) Secretion Confirmation

The 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 incubated 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 in FIG. 2A, the compounds of the present invention inhibited HBsAg secretion by 50% or more at a concentration of 10 μM. NJK14021 (Example 9) and NJK14027 (Example 10) showed about 50% inhibition at 2 [mu] M.

In particular, NJK14047 (Example 11) showed the greatest inhibition of p38 MAPK and HBsAg secretion. As shown in Fig. 2 B, in vitro p38 MAPK inhibitor and HBsAg secretion inhibition by the compound is represented by the high positive correlation (r ² = 0.8547) that the compound can inhibit the HBsAg secretion through the p38 MAPK inhibition Respectively.

&Lt; Experimental Example 3 >

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

Cytotoxicity was confirmed by cytotoxicity of NJK14047 on various cells using MTT (3- (4-5) -dimenthylthiazol-2-yl) -2,5-diphenyltetrazolium bromide analysis. Briefly, HepG2, HepG2.2.15, AML12, A549 or HeLa cells were cultured in 96-well plates 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 were cultured in AML12 medium containing GlutaMAX, insulin (0.005 mg / mL), transferrin (0.005 mg / mL), selenium (5 ng / mL), dexamethasone (40 ng / mL) (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 further cultured for 4 hours. After aspiration, DMSO was added to dissolve the cells and dissolve the water-insoluble formazan. Absorbance at 570 nm was measured with a microplate reader.

The NJK14047 compound did not show significant cytotoxicity in HepG2.2.15 cells at concentrations up to 20 μM. Therefore, 20 [mu] M of NJK14047 was used at the maximum concentration in the following experimental examples (see Fig. 3A).

In addition, the cytotoxic effect of NJK14047 on other cell lines was further analyzed. As shown in FIG. 3B, the 50% cytotoxic concentration (CC ₅₀ ) value ranged from 53.7 to 110.4 μM, and the CC ₅₀ value of NJK14047 in normal mouse hepatic cells (AML-12 cells) (125.99 [mu] M).

EXPERIMENTAL EXAMPLE 4 Confirmation of p38 MAPK Inhibition in Hepatocytes

To confirm p38 MAPK inhibition in hepatocytes, HepG2 cells transformed with plasmid containing HBV genome (pHBV-112 1.2x, GenBank no. AY641558) were treated with 5 or 20 μM NJK14047 (Example 11) and immunoblotted Respectively. Treatment with NJK14047 reduced p38 MAPK phosphorylation without affecting total protein levels, indicating that NJK14047 could inhibit p38 MAPK activation in HBV infected cells. (See FIG. 3C).

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

<Experimental Example 5> Measurement of HBsAg and HBeAg secretion

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

As shown in Figure 4A NJK14047 inhibited the secretion of HBsAg in a dose-dependent manner from the HepG2.2.15 cells (IC ₅₀ = 5.3 μM, Fig. 4A). The effect of NJK14047 on HBeAg secretion with HepG2.2.15 cells could not be detected and was also determined by ELISA (data not shown). The antiviral effect of NJK14047 was confirmed using an HBV genomic transduced model with the genotype C virus genome.

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

&Lt; Experimental Example 6 >

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

Viral pellets from HBV genome-injected cells were treated with RQ1 DNase I (Promega, Wis., USA) according to the manufacturer's instructions to remove residual DNA used for transfection. The 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 treatment with NJK14047, indicating that NJK14047 could inhibit viral replication as well as viral replication and viral particle production (see Figure 4D). Similar results were derived from experiments with HepG2 cells transfected with pHBV-1.2x (see FIG. 4E).

<Experimental Example 7> Measurement of pgRNA production

The effect of the compound of the present invention NJK14047 (Example 11) on HBV pgRNA production was analyzed. HBV pgRNA production in the cells was slightly altered (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 glycican 5 as an entry factor for hepatitis B and D viruses. : 35-48.). Total RNA was isolated from HepG2.2.15 or HepG2 cells transfected with HBV genome using TRIzol and treated with RQ1 DNase I (Promega). The RNA was purified by extraction with conventional phenol-chloroform isoamyl alcohol and ethanol precipitation, and cDNA synthesis was performed with an RNA sample (1.).

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 with GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) as a control.

Treatment of HepG2.2.15 cells (FIG. 5A) and pHBV-1.2x-transfected HepG2 cells (FIG. 5B) for 48 hours in the amount of NJK14047 or entecavir (ETV, 30 nM) resulted in 3.5 kb pgRNA &Lt; / RTI &gt; was observed compared to in vehicle-treated cells. .

<Experimental Example 8> Quantification of HBV nuclear cccDNA

HBV cccDNA of HepG2.2.15 or HepG2 cells transfected with the linearized HBV genome were quantitated by partial modification (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 epigenetic regulation of the nuclear ccc DNA minichromosome. J Clin Invest 122: 529-537.). The collected cells were washed with cold phosphate buffered saline and cultured on ice for 10 minutes with 270 μl of lysis buffer (50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid [EDTA], 1% NP-40) .

After the brief centrifugation (10,000 x g, 1 min), the pelleted nuclei were digested with digestion buffer B (10 mM Tris-HCl, 10 mM EDTA, 150 mM NaCl, 0.5% sodium dodecyl sulfate, and 0.5 mg / And incubated overnight at &lt; RTI ID = 0.0 &gt; 37 C. &lt; / RTI &gt;

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

cccDNA was purified using a 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 '.

As shown in FIGS. 5C and 5D, treatment with NJK14047 revealed a significant decrease in 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 that the p38MAPK inhibition by the compound NJK14047 of the present invention can inhibit cccDNA production and pgRNA synthesis.

Experimental Example 9 In Vitro HBV Infection Assay

The culture supernatant was collected from HepG2.2.15 cells (4 x 107 cells / 150-mm dish) every two days for 14 days. The supernatant was briefly centrifuged and filtered to remove cell debris (0.45 mu m).

The virus particles were concentrated by ultracentrifugation (100,000 x g, 3 hours, 4 占 폚). 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 24 hours of incubation, the cells were washed and maintained in AML12 medium containing NJK14047 for 7 days. Extracellular viral DNA and HBsAg levels were determined by qPCR (quantitative PCR) and ELISA as described above.

As a result, the NJK14047 treatment showed a dose-dependent reduction in HBV virion particle production (both virion particle production) and HBsAg secretion (see FIGS. 6A and B).

Therefore, the compound of formula (I) according to the present invention reduces the secretion of HBsAg and the secretion of HBeAg, reduces the level of HBV virus, inhibits pgRNA production and inhibits cccDNA production, Or pharmaceutical compositions for therapeutic use.

Meanwhile, the compound represented by Formula 1 according to the present invention can be formulated into various forms according to the purpose. Hereinafter, some formulating methods in which the compound represented by Formula 1 according to the present invention is contained as an active ingredient are exemplified, and the present invention is not limited thereto.

&Lt; Formulation Example 1 > Preparation of powders

The compound of formula (1) 2 g

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

&Lt; Formulation Example 2 > Preparation of tablet

The compound of formula (1) 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

&Lt; Formulation Example 3 > Preparation of capsules

The compound of formula (1) 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; Formulation Example 4 > Preparation of injection

The compound of formula (1) 100 mg

Mannitol 180 mg

Na2HPO4. 2H2O 26 mg

Distilled water 2974 mg

According to the conventional method for preparing an injectable preparation, an injectable preparation was prepared by incorporating the aforementioned components in the amounts indicated.

Manufacturing Example of Health Functional Food

The compound represented by Formula 1 according to the present invention can be manufactured into various types of health functional foods according to the purpose. Hereinafter, the method of manufacturing some health functional foods containing the active substance according to the present invention as an active ingredient is exemplified, and the present invention is not limited thereto.

&Lt; Health functional food production example 1 > Production of health functional food

The compound of formula (1) 100 mg

Vitamin mixture Suitable amount

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

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture Suitable amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a component suitable for a health functional food as a preferred embodiment, the compounding ratio may be arbitrarily changed, and the above components may be mixed , Granules may be prepared and used in the manufacture of health functional food compositions according to conventional methods.

&Lt; Health Functional Food Production Example 2 >

The compound of formula (1) 100 mg

Citric acid 100 mg

oligosaccharide 100 mg

Plum concentrate 2 mg

Taurine 100 mg

Purified water is added to the entire 500 mL

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized container. The resulting solution was sealed and sterilized, &Lt; / RTI &gt; Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (10)

1. A compound represented by the following formula (1): &lt; EMI ID =
[Chemical Formula 1]

(In the 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 according to claim 1,
R ² is H, C _2-7 straight or branched chain alkyl, or phenyl C _1-3 straight chain or branched chain alkyl, or a pharmaceutically acceptable salt thereof.
The method according to claim 1,
R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl, or a pharmaceutically acceptable salt thereof.
The method according to claim 1,
Wherein the compound represented by the formula (1) is any one selected from the group consisting of the following 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,
A step of dissolving Compound 2 and Compound 3 in a pyridine solvent and reacting at room temperature to obtain Compound 1 (Step 1);
(1), wherein R &lt; 1 &gt; and R &lt; 2 &gt;
[Reaction Scheme 1]

(In the above Reaction Scheme 1,
Wherein R &lt; ¹ &gt; and R &lt; ² &gt; are the same as defined in formula (1).
A pharmaceutical composition for the prevention or treatment of hepatitis B comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
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 &lt; ³ &gt; is _C1-5 straight chain or branched alkoxy, and the _C1-5 linear or branched alkoxy may be substituted with at least one hydroxy group.
The method according to claim 6,
R ¹ is

or

ego;
R ² is H, C _2-7 linear or branched alkyl, or phenyl C _1-3 linear or branched alkyl;
R ³ is H, or C _1-4 straight chain or branched alkoxy, and wherein the C _1-4 straight chain or branched alkoxy may be substituted with one to four hydroxy groups.
The method according to claim 6,
R ¹ is

or

ego;
R ² is H, C _4-5 straight or branched chain alkyl, or phenylmethyl;
R &lt; ³ &gt; is H, methoxy, or 2,3-dihydroxypropyl.
The method according to claim 6,
Wherein the compound represented by the formula (1) is any one selected from the group consisting of the following 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;
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; And
11) N-Cyclopropyl-4 '- (4- (2,3-dihydroxypropoxy) benzoyl) -6-methyl- [1,1'-biphenyl] -3-carboxamide.
A health functional food composition for preventing or ameliorating hepatitis B comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
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 &lt; ³ &gt; is _C1-5 straight chain or branched alkoxy, and the _C1-5 linear or branched alkoxy may be substituted with at least one hydroxy group.

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