KR20150065130A - Benzidine derivative, preparation method thereof and pharmaceutical composition for treating liver disease related with Hepatitis C virus containing the same - Google Patents

Abstract

The present invention relates to: a compound having anti-virus activity against hepatitis C virus; an optical isomer or pharmaceutically acceptable salt of the same; method for producing the same; and a pharmaceutical composition containing the same as an active ingredient for preventing or treating liver disease caused by hepatitis C virus. According to the present invention, a benzidine derivative has excellent anti-virus activity against hepatitis C virus and exhibits excellent pharmacological activity in a human body. Therefore, the benzidine derivative can be advantageously used as a pharmaceutical composition for preventing or treating liver disease such as acute hepatitis C, chronic hepatitis C, cirrhosis, and hepatocellular cancer which are caused by hepatitis C virus.

Description

[0001] The present invention relates to a benzidine derivative, a process for producing the same, and a pharmaceutical composition for treating liver disease by the hepatitis C virus containing the same,

The present invention relates to a compound having an antiviral activity against a C-type virus, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, a method for preparing the same, and a method for preventing or treating liver disease caused by hepatitis C virus ≪ / RTI >

Hepatitis C virus (HCV) is an RNA virus classified as a hepacivirus genus of the Flaviviridae, which was discovered in the United States in 1989.

It is reported that the hepatitis C virus (HCV) infection is caused by transfusion and community-acquired, and about 70% is infected by kidney dialysis. It is known that about 20% of hepatitis C virus infections cause acute hepatitis accompanied by cirrhosis within 5 years and metastasis to liver cancer (Davis et al, New. Engl. ) 1501 / Alter et al, Leonard et al., Current Pespective in Hepatology, (1989) p.83). This high rate of chronic infection is an uncommon event in RNA viruses and is evidence that hepatitis C virus is a mediator of high rates of liver cancer. In recent years, hepatitis C virus (HBV) has been successfully screened for all blood, and infection due to blood transfusion has been significantly reduced. However, since local infection is not effectively managed and infection rate is high, .

Epidemiologically, unlike hepatitis B virus (HBV), hepatitis C virus is spread all over the world and it is reported that 1.5% to 2% of the world's population is infected. Hepatitis C virus infection is characterized by progression to chronic hepatitis. The probability of liver metastasis and liver metastasis is significantly higher than that of hepatitis B virus. Because hepatitis C virus is taxonomically different from the hepatitis B virus, it can not be prevented by the hepatitis B virus vaccine. In addition, interferon and ribavirin, an antiviral agent, are currently used as a therapeutic agent for hepatitis C virus infection (Hayashi N., et al., J. Gastroenterol., 41, (2006) The response is remarkably different depending on the genotype, the effect is extremely weak, and the side effects of the two drugs used in combination therapy are large and expensive. Therefore, development of a more effective new hepatitis C virus agent is required.

In order to overcome the above-mentioned problems, the hepatitis C virus (HCV) has been characterized to express the pharmacological activity of hepatitis C virus (HCV) by blocking certain stages of the life cycle of the hepatitis C virus.

The life cycle of hepatitis C virus is as follows. HCV attached to the surface of the host cell invades into the host cell by endocytosis. Thereafter, a precursor protein consisting of about 3,000 amino acid residues is generated from the hepatitis C virus genome RNA that has invaded into the host cell. The hepatitis C virus genome or the NS3 and NS4 protease encoded by the signal peptidase of the host cell to produce capsid protein, envelope protein, NS3 and NS4 protease, NS5B About 10 virus proteins such as RNA polymerase are produced. Genomic RNA cloned by NS 2B polymerase binds to capsid proteins and envelope proteins mediated by a-glucosidase and becomes viral particles. Hepatitis C virus particles are released from host cells (Manns MP, et al., Nat. Rev. Drug. Discov., 6, (2007), 991).

Agents that exhibit hepatitis C virus-inhibiting activity by blocking certain stages of the hepatitis C virus life cycle as described above may be classified as either an RNA polymerase inhibitor-type, a protease inhibitor-type, an a- glucosidase inhibitor- And other types. For example, RNA polymerase inhibitor types such as MK-7009 (Merck) and R7128 (Pharmasset / Roche) are on Phase 1 of clinical trials and include VCH-759 (Virochem), R1626 (Roche), valopicitabine Idenix) is on clinical trial 2. In addition, among the protease inhibitor types, ITMN-191 (R-7227, InterMune / Roche) is on clinical trial 1, TMC435350 (Medivir / Tibotec) is on clinical trial 2, Boceprevir (SCH 503034), Schering ) And Telaprevir (Vertex) are on Phase 3 of the trial. In addition, the cyclophilin inhibitor type DEBIO-025 (DEBIO) and the glucosidase I inhibitor type celgosivir (MIGEBIX) are on clinical trial 2 (Kronenberger B., et al., Clin Liver Dis. 12, (2008), 529).

However, since the emergence of viruses resistant to the hepatitis C virus infection, which has undergone clinical trials, has already been reported, a novel C type hepatitis virus Development of hepatitis virus agent is urgently required.

Accordingly, the present inventors have studied a hepatitis C virus agent having low cytotoxicity and excellent antiviral activity against hepatitis C virus, and have found that benzidine derivatives have excellent antiviral activity against hepatitis C virus, And completed the present invention.

It is an object of the present invention to provide a compound having an antiviral activity against hepatitis C virus, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

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 liver disease caused by hepatitis C virus containing the above compound as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or ameliorating liver disease caused by hepatitis C virus containing the above compound as an active ingredient.

In order to achieve the above object,

There is provided a compound represented by the following formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

R ¹ and R ² are independently -H, -OH, a halogen, C _{1 -10} straight or branched chain alkyl, C _{1 -10} linear or branched alkoxy, unsubstituted or substituted C _{6 -10} of the aryl, or - NHC (= O) R < ¹² >

Here, the above substituted aryl of _{6 -10} C is one or more substituents selected from the group consisting of straight or branched alkyl, straight-chain or branched alkoxy and halogen of C _{1 -5} C for _{1 -5} may be substituted,

In addition, the R ¹² is -H, or a linear or branched alkoxy of ₁ C and _-5;

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently selected from -H, halogen, unsubstituted or substituted C _{1 -5} straight or branched ego,

Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring of _{5 -6} C with the carbon atoms to which they are connected, the ring include halogen, C ₁ of the C _{5 -6 -5} straight or branched chain alkyl and = O may be substituted;

R ¹¹ is -H, -OH, halogen, straight or branched chain alkoxy of C _{1 -10} straight or branched chain alkyl, C _{1 -10} of, or is = O;

는 단일 또는 이중결합이고;

Is a single or double bond;

a is an integer of 0-3.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound represented by formula (2) with a compound represented by formula (3) in an organic solvent to prepare a compound represented by formula (4) (step 1);

Removing the protecting group of the compound represented by the formula (4) prepared in the step (1) to prepare a compound represented by the formula (5) (step 2); And

Reacting a compound represented by the formula (5) and a compound represented by the formula (6) to prepare a compound represented by the formula (1) (step 3). A method for producing the compound is provided.

[Reaction Scheme 1]

(In the above Reaction Scheme 1,

는 상기 화학식 1에서 정의한 바와 같고; ^{R 1, R 2, R 3} , R 4, R 5, R 6, R 7, R 8, R 9, R 10, R 11, a, and

Is as defined in Formula 1;

PG is a protecting group).

Further, the present invention provides a pharmaceutical composition for preventing or treating liver disease caused by hepatitis C virus comprising the compound represented by the above-mentioned formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating liver disease caused by hepatitis C virus comprising the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient .

The benzidine derivative according to the present invention exhibits excellent antiviral activity including the activity of the viral agent for hepatitis C virus, has no toxicity to cells, exhibits excellent pharmacological activity in vivo, and is free from toxicity in heart and plasma The pharmaceutical composition containing it as an active ingredient is useful as a pharmaceutical composition for the prevention or treatment of liver diseases such as acute hepatitis C, chronic hepatitis C, cirrhosis and hepatocellular carcinoma caused by hepatitis C virus Can be used.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following general formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

R ¹ and R ² are independently -H, -OH, a halogen, C _{1 -10} straight or branched chain alkyl, C _{1 -10} linear or branched alkoxy, unsubstituted or substituted C _{6 -10} of the aryl, or - NHC (= O) R < ¹² >

Here, the above substituted aryl of _{6 -10} C is one or more substituents selected from the group consisting of straight or branched alkyl, straight-chain or branched alkoxy and halogen of C _{1 -5} C for _{1 -5} may be substituted,

In addition, the R ¹² is -H, or a linear or branched alkoxy of ₁ C and _-5;

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently selected from -H, halogen, unsubstituted or substituted C _{1 -5} straight or branched ego,

Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring of _{5 -6} C with the carbon atoms to which they are connected, the ring include halogen, C ₁ of the C _{5 -6 -5} straight or branched chain alkyl and = O may be substituted;

R ¹¹ is -H, -OH, halogen, straight or branched chain alkoxy of C _{1 -10} straight or branched chain alkyl, C _{1 -10} of, or is = O;

는 단일 또는 이중결합이고;

Is a single or double bond;

a is an integer of 0-3.

Preferably,

R ¹ and R ² are independently -H, -OH, a halogen, C _{1 -5} straight or branched chain alkyl, C _{1 -5} straight or branched chain alkyl, unsubstituted or substituted C _{6 -8} in the aryl, or - NHC (= O) R < ¹² >

Wherein said substituted C _{6 -8} aryl may be substituted with one or more substituents selected from the group consisting of C _{1 -3} straight or branched alkyl, C _{1 -3} straight or branched alkoxy and halogen,

Also, R ¹² is -H or C _{1 -3} straight or branched alkoxy;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently selected from -H, halogen, unsubstituted or substituted C _{1 -3} straight or branched ego,

Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring of _{5 -6} C with the carbon atoms to which they are connected, the ring include halogen, C ₁ of the C _{5 -6 -3} straight or branched chain alkyl and = O may be substituted;

R ¹¹ is -H, -OH, halogen, C _{1 -5} straight or branched chain alkyl, C _{1 -5} straight or branched alkoxy, or = O;

는 단일 또는 이중결합이고;

Is a single or double bond;

a is an integer of 0-2.

More preferably,

R ¹ and R ² are independently phenyl, or methoxycarbonylamino;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently -H, -F, -Cl, -Br, -CF ₃ ,

Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring in C ₅ along with the carbon atoms to which they are connected together, with -F, = O, and methyl, the ring of the C ₅ One or more substituents selected from the group consisting of the substituents may be substituted;

R < ¹¹ > is -H, or = O;

는 단일 또는 이중결합이고;

Is a single or double bond;

a is an integer of 0-1.

The compound represented by Formula 1 may be any one selected from the following group of compounds.

(1) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;

(2) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'-bis (trifluoromethyl) - [ (2-oxo-1-phenylethan-2-yl) -4,4'-diyl) bis (azodioyl)) bis (carbonyl)) bis (pyrrolidin- 1-yl))) dicarbamate;

(3) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;

(4) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' Carbonyl)) bis (pyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(5) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl)) bis (carbonyl)) bis (pyrrolidine- )) Dicarbamate;

(6) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;

(7) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;

(8) Synthesis of dimethyl ((1R, 1'R) - ((2R, 2'R) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate; bis (pyrrolidine-2,1-diyl)) bis

(9) Synthesis of dimethyl ((1R, 1'R) - ((5S, 5'S) -5,5 ' )) Bis (carbonyl)) bis (3-oxopyrrolidin-5,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(10) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (piperidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(11) Synthesis of dimethyl ((1R, 1'R) - ((2R, 2'R) -2,2 ' Bis (carbonyl)) bis (piperidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(12) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (2-methylpyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(13) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (Azodioyl)) bis (carbonyl)) bis (pyrrolidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;

(14) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' Bis (carbonyl)) bis (pyrrolidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate; And

(15) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (pyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate.

The compound represented by the formula (I) according to the present invention can be used in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, and the like. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dynitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluene sulfonate, claw Benzene sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene- Sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be obtained by a conventional method, for example, by dissolving the compound represented by the formula (1) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, Followed by filtration and drying. Alternatively, the solvent and excess acid may be distilled off under reduced pressure, followed by drying or crystallization in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

The present invention includes not only the compound represented by the formula (1) and pharmaceutically acceptable salts thereof, but also possible solvates, hydrates, optical isomers and the like which can be prepared therefrom.

Since the compound represented by Formula 1 according to the present invention is excellent in antiviral activity against hepatitis C virus, the pharmaceutical composition comprising the compound as an active ingredient can be used for the treatment of acute hepatitis C, chronic hepatitis C Hepatitis, liver cirrhosis, hepatocellular carcinoma, and the like.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound represented by formula (2) with a compound represented by formula (3) in an organic solvent to prepare a compound represented by formula (4) (step 1);

Removing the protecting group of the compound represented by the formula (4) prepared in the step (1) to prepare a compound represented by the formula (5) (step 2); And

(Step 3), which comprises reacting the compound represented by Chemical Formula 5 and the compound represented by Chemical Formula 6, prepared in Step 2, to prepare a compound represented by Chemical Formula 1 .

[Reaction Scheme 1]

(In the above Reaction Scheme 1,

는 상기 화학식 1에서 정의한 바와 같고; ^{R 1, R 2, R 3} , R 4, R 5, R 6, R 7, R 8, R 9, R 10, R 11, a, and

Is as defined in Formula 1;

PG is a protecting group).

Hereinafter, the process for preparing the compound represented by the formula (1) according to the present invention will be described in detail.

In the process for preparing a compound represented by the formula (1) according to the present invention, the step (1) is carried out by reacting the compound represented by the formula (2) and the compound represented by the formula (3) together with an amide reagent in an organic solvent, More specifically, an amidation reaction of an amine group of a compound represented by the formula (2) and a carboxylic acid group of a compound represented by the formula (3) is carried out in the presence of an amidating agent to obtain a coupling product represented by the formula Is a step of preparing a compound represented by the formula

The amide reagent is then reacted with diisopropylethylamine (DIPEA), triethylamine (TEA) or dimethylaminopyridine (DMAP) in the presence of benzotriazol-1-yl-oxy- (Methyl-amino) -phosphonium hexafluorophosphate (Py-BOP), O-benzotriazole-N, N, N, N-tetramethyl-euenium-hexafluoro-phosphate (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (HAT), and the like. Ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), and carbonyldiimidazole (CDI) -Dimethylaminopropyl) carbodiimide (EDC) can be used.

The organic solvent may be used alone or in combination with methanol, dimethylformamide, tetrahydrofuran, dichloromethane, toluene or the like, preferably dichloromethane.

Further, the reaction is preferably carried out at a temperature of 0 ° C between the boiling point of the solvent, and the reaction time is not particularly limited, but preferably 0.5-10 hours.

In the process for preparing a compound represented by the formula (1) according to the present invention, the step (2) is a step for preparing a compound represented by the formula (5) by removing the protecting group of the compound represented by the formula (4) Specifically, the amine protecting group (PG) contained in the compound represented by the formula (4) prepared in the step 1 is deprotected using a deprotecting agent to prepare a compound represented by the formula (5) wherein the amine protecting group is deprotected .

Examples of the amine protecting group (PG) contained in the compound of Formula 3 include t-butoxycarbonyl (Boc), 9H-fluoren-9-ylmethoxycarbonyl (Fmoc), trityl, Benzyl, chloroacetyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, formyl, trifluoroacetyl, p-toluenesulfonyl, benzenesulfonyl, methanesulfonyl, p- nitrobenzyloxycarbonyl, 2,2-trichloroethoxycarbonyl, allyloxycarbonyl (Alloc) and the like can be used, and t-butoxycarbonyl (Boc) can be preferably used.

In addition, the deprotection conditions of the step 2 according to the present invention can use an unprotected condition that is conventionally used according to a protected protecting group.

Further, the reaction is preferably carried out at a temperature of 0 ° C between the boiling point of the solvent, and the reaction time is not particularly limited, but preferably 0.5-10 hours.

In the process for preparing the compound represented by the formula (1) according to the present invention, the compound represented by the formula (5) and the compound represented by the formula (6) prepared in the step 2 are reacted together with the amidating agent to produce a compound represented by the formula More specifically, an amidation reaction of an amine group of the compound represented by the general formula (5) deprotected in the step 2 and a carboxylic acid group represented by the general formula (6) in the presence of an amidating agent is carried out to obtain a compound represented by the general formula Is a step of preparing a compound represented by the formula

The amide reagent is then reacted with diisopropylethylamine (DIPEA), triethylamine (TEA) or dimethylaminopyridine (DMAP) in the presence of benzotriazol-1-yl-oxy- (Methyl-amino) -phosphonium hexafluorophosphate (Py-BOP), O-benzotriazole-N, N, N, N-tetramethyl-euenium-hexafluoro-phosphate (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (HAT), and the like. Ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), and carbonyldiimidazole (CDI) -Dimethylaminopropyl) carbodiimide (EDC) can be used.

The organic solvent may be used alone or in combination with methanol, dimethylformamide, tetrahydrofuran, dichloromethane, toluene or the like, preferably dichloromethane.

Further, the reaction is preferably carried out at a temperature of 0 ° C between the boiling point of the solvent, and the reaction time is not particularly limited, but preferably 0.5-10 hours.

The present invention also provides a pharmaceutical composition for preventing or treating liver disease caused by hepatitis C virus comprising the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the pharmaceutical composition according to the present invention, the liver disease caused by the hepatitis C virus includes acute hepatitis C, chronic hepatitis C, cirrhosis, hepatocellular carcinoma and the like.

Thus, the anti-C virus activity of the compound represented by formula (1) according to the present invention was evaluated. Specifically, when the compound of Chemical Formula 1 according to the present invention was treated at a concentration of 1 μM in hepatocarcinoma cells inoculated with hepatitis C virus containing the luciferase gene, the hepatitis C virus contained low concentrations of the compound represented by Chemical Formula 1 (See Experimental Example 1).

In addition, it was confirmed that the EC ₅₀ value indicating the antiviral activity was low in the experiment in which the compound represented by Chemical Formula 1 according to the present invention was treated to the copied hepatitis C virus. Especially, in Examples 2, 3, 4, 5, 6, 7, 9, 13, 14 and 15 were found to have very low EC ₅₀ values, respectively (see Experimental Example 2). From this, it can be seen that the compound represented by the formula (I) according to the present invention has excellent antiviral activity against hepatitis C virus.

Specifically, the compound represented by formula (I) according to the present invention acts as a target for NS5A (Nonstructural protein 5A), which is known as a factor necessary for RNA replication of a virus. The NS5A is a 56 kDa protein in the form of phosphorylation involved in HCV RNA replication. It has an amphipathic alpha helix structure that catalyzes attachment to the host cell membrane, (Cys 39, 57, 59 and 80), binding with PI3K, PKR and NS5B, and viruses.

Known inhibitors known as NS5A include ACH-2928, AZD-7295, PPI-461, BMS 824393, GS-5885 and Vertex.

Since the NS5A has a unique zinc binding site, the compound represented by the formula (1) according to the present invention binds to the above position and acts as an inhibitor of NS5A, thereby exhibiting an antiviral activity against hepatitis C virus And there is a need for further research to be carried out in the future.

Further, the cytotoxicity of the compounds of Examples 1 to 12 according to the present invention was evaluated. As a result, it was confirmed that when treated at a concentration of 1 μM, there was no cytotoxicity. From this, it can be seen that the compound represented by Chemical Formula 1 according to the present invention has low cytotoxicity (see Experimental Example 3).

Therefore, the compound represented by the formula (I) according to the present invention is excellent in antiviral activity against hepatitis C virus and has no cytotoxicity. Therefore, the pharmaceutical composition containing the compound as an active ingredient is an acute C Hepatitis, chronic hepatitis C, liver cirrhosis, hepatocellular carcinoma, and the like.

When the pharmaceutical 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 can be administered orally or parenterally But the present invention is not limited thereto.

Examples of formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules, elixirs and troches, (E.g., silica, talc, stearic acid or a magnesium salt or a calcium salt 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, optionally mixed with starch, agar, alginic acid or its sodium salt The same disintegrating or boiling mixture and / or absorbing agent, coloring agent, flavoring agent and sweetening agent.

The pharmaceutical composition containing the compound represented by the formula (1) as an active ingredient may be administered parenterally, and the parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

In this case, in order to formulate the formulation for parenteral administration, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof may be mixed with water or a stabilizer or a buffer to prepare a solution or suspension, . The compositions may contain sterilized and / or preservatives, stabilizers, wettable or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, Or may be formulated according to the coating method.

The dosage of the pharmaceutical composition containing the compound represented by the formula (1) as an active ingredient may vary depending on the patient's age, body weight, sex, dosage form, health condition and disease severity, 200 mg / kg / day, may be administered orally or parenterally by dividing a predetermined time interval several times a day, preferably once or three times a day, according to the judgment of a doctor or pharmacist.

The present invention also provides a health functional food composition for preventing or ameliorating liver disease caused by hepatitis C virus comprising the compound represented by the formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient .

In the health functional food composition according to the present invention, the liver diseases caused by the hepatitis C virus include acute hepatitis C, chronic hepatitis C, cirrhosis, hepatocellular carcinoma and the like.

The health functional food composition according to the present invention can be added to health functional foods such as food, beverage, etc. for the purpose of preventing or improving liver disease caused by hepatitis C virus.

There is no particular limitation on the kind of the food. Examples of the foods to which the above substances can be added include dairy products including dairy products, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, Beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, all of which include health functional foods in a conventional sense.

The compound represented by formula (1) according to the present invention can be used as it is in food or in combination with other food or food ingredients, and can be suitably used according to conventional methods. 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 compound represented by the formula (1) in the health functional food may be 0.1 to 90 parts by weight based on the total weight of the food. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, 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 beverage of the present invention is not particularly limited to the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Daisakaride, for example Maltose, sucrose etc; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavors (e.g., tau martin, stevia extract, etc.) and synthetic flavorings (e.g., saccharin, aspartame, etc.) can be advantageously used as flavorings 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 healthy function foods of the present invention.

In addition to the above, the health functional food composition containing the compound represented by the formula (1) according to the present invention as an active ingredient may be used as a nutritional supplement, a vitamin, a mineral (electrolyte), a flavor such as a synthetic flavor and a natural flavor, (For example, cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, ≪ / RTI > 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 ratio 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 according to the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples.

It should be noted, however, that the following examples and experimental examples are illustrative of the present invention, and the present invention is not limited by the following examples and experimental examples.

< Manufacturing example  1 > (R) -2- ( Methoxycarbonylamino ) -2-phenylacetic acid

Sodium carbonate (0.55 g, 5.2 mmol) was added to an aqueous sodium hydroxide solution (10 ml, 10 mmol) in which D-phenylglycine (1.5 g, 10 mmol) was dissolved and cooled using an ice bath. Methyl chloroformate (0.85 ml, 11.0 mmol) was slowly added dropwise to the cooled reaction mixture. When the dropwise addition was complete, the ice bath was removed and the mixture was stirred at room temperature for 3.25 hours. The reaction product was then washed three times with ether (18 ml) and the water layer was cooled in an ice bath and acidified to pH 1-2 with hydrochloric acid (conc. HCl). The acidified aqueous layer was filtered and was extracted three times with dichloromethane (18 ml), drying the extracted organic layer with magnesium sulfate (MgSO _4). The filtered organic layer was concentrated under reduced pressure, and the obtained oil residue was treated with diethyl ether / hexane (~5: 4 ratio; 10 ml) to obtain a precipitate. The precipitate obtained above was washed with diethyl ether / hexane (ratio of 1: 3), filtered and dried under vacuum to obtain the desired compound (1.4 g, 67%) as a white solid.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 500 MHz): 12.79 (br s, 1H), 7.96 (d, J = 12, 1H), 7.40-7.29 (m, 5H), 5.13 (d, J = 12, 1H), 3.55 (s, 3H).

< Manufacturing example  2> 9,9-Dimethyl-9 H - Fluorene -2,7- Diamine  Produce

Step 1: 9,9-Dimethyl-2,7- Dinitro -9 H - Fluorene  Produce

2,7-nitro -9 H - in a mixture of fluorene (100 mg, 0.39 mmol) and NaOt-Bu (75 mg, 0.78 mmol) N, was dissolved in DMF in an ice bath. Iodomethane (49 mL, 0.78 mmol) was slowly added to the mixture, and after mixing for 2 hours, water was added to obtain a precipitate. The precipitate was filtered, washed with water, and then dried. Without further purification, the desired product (89 mg, 80%) was obtained in the form of a yellow solid.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 8.59 (d, 2H), 8.33 (m, 4H), 1.60 (s, 6H).

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 156.2, 148.1, 142.7, 123.6, 122.9, 118.7, 47.9, 25.6.

Step 2: 9,9-Dimethyl-9 H - Fluorene -2,7- Diamine  Produce

Fe ₃ O ₄ (15 mg, 0.063 mmol) and DMF (1.9 mL) were placed in an oven-dried Schlenk tube and heated in an ultrasound bath in argon for 1 minute Sonicate. With 9,9-dimethyl-2,7-di nitro obtained in Step 1 -9 H - was added to the fluorene (90 mg, 0.32 mmol) and hydrazine monohydrate (123 mL, 2.52 mmol) to the mixture. The reaction mixture was mixed at 80 &lt; 0 &gt; C in argon until the reaction was terminated. After magnetic separation using a catalyst, the organic layer was concentrated in vacuo. The residue was separated by CH ₂ Cl ₂ and H ₂ O. After washing the organic layer with brine, dried with MgSO _4, filter, and concentrated in vacuo. Without further purification, the desired product (69 mg, 98%) was obtained in the form of a yellow solid.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 7.21 (s, 1H), 7.19 (s, 1H), 6.6 (d, 2H), 6.47 (d, 1H), 6.45 (d, 1H), 4.9 (s, 4H), 1.3 (s, 6H).

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 153.5, 146.7, 128.4, 118.6, 112.6, 108.5, 45.6, 27.7.

_{^{19 F NMR (DMSO- d 6,}} 377 MHz,): δ-106.7. LC / MS: Anal. Calcd.

^{For [M + H] + C} 15 H 16 N 2: 225.1386; found 225.1383.

< Manufacturing example  3> 9,9- Difluoro -9 H - Fluorene -2,7- Diamine  Produce

Step 1: 9,9- Difluoro -2,7- Dinitro -9 H - Fluorene  Produce

A 2,7 nitro -9 H - fluorene (100 mg, 0.39 mmol) and a N- fluorobenzene sulfonimide (N-Fluorobenzenesulfonimide, NFSI) ( 369 mg, 1.17 mmol) was dissolved in the DMF, -20 Lt; 0 &gt; C. NaHMDS (1.0 M in THF, 1.17 mL, 1.17 mmol) was added dropwise over 5 minutes. After 2 hours at 0 &lt; 0 &gt; C, the reaction was terminated by TLC and MeOH was added to quench excess base. The suspension was filtered using celite and concentrated in vacuo. A silica gel mesh was prepared from the residue and the desired product (22 mg, 19%) was obtained in the form of a yellow solid via flash chromatography (silica gel: EtOAc / hexane as eluent).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 8.63 (d, 2H), 8.53 (d, 1H), 8.56 (d, 1H), 8.36 (s, 1H), 8.34 (s, 1H).

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 149.1, 142.5, 138.5, 129.2, 144.3, 120.8, 119.6.

_{^{19 F NMR (DMSO-d 6}} , 377 MHz,): δ-110.3.

Step 2: 9,9- Difluoro -9 H - Fluorene -2,7- Diamine  Produce

Fe ₃ O ₄ (4 mg, 0.015 mmol) and DMF (0.5 mL) were placed in an oven-dried Schlenk tube and heated in an ultrasound bath in argon for 1 minute Sonicate. A-2,7-nitro-9,9-difluoro obtained in Step 1 -9 H - it was added to the fluorene (22 mg, 0.075 mmol) and hydrazine monohydrate (29 ㎕, 0.60 mmol) to the mixture . The reaction mixture was mixed at 80 &lt; 0 &gt; C in argon until the reaction was terminated. After magnetic separation using a catalyst, the organic layer was concentrated in vacuo. The residue was separated by CH ₂ Cl ₂ and H ₂ O. After washing the organic layer with brine, dried with MgSO _4, filter, and concentrated in vacuo. Without further purification, the desired product (17 mg, 98%) was obtained in the form of a yellow solid.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 7.19 (s, 1H), 7.17 (s, 1H), 6.8 (d, 2H), 6.61 (d, 1H), 6.59 (d, 1H), &lt; / RTI &gt; 5.3 (s, 4H).

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 148.0, 137.3, 137.1, 127.7, 123.7, 119.8, 116.45, 109.1.

_{^{19 F NMR (DMSO-d 6}} , 377 MHz,): δ-106.7.

LC / MS: Anal. Calcd. ^{For [M + H] + C} 13 H 10 F 2 N 2: 233.0885; found 233.0885.

< Manufacturing example  4> 2,7- Diamino -9 H - Fluorene -9-one

A mixture of 9 H -fluorene-2,7-diamine (294 mg, 1.5 mmol) and Cs ₂ CO ₃ (1.5 g, 4.5 mmol) was dissolved in DMSO (7 mL) air. After confirming that all of the reaction materials had disappeared through TLC, water was added to the solution to obtain a precipitate. The precipitate was filtered, washed with water, and then dried. The desired product (239 mg, 76%) was obtained in solid form without further purification.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 7.10 (s, 1H), 7.08 (s, 1H), 6.70 (d, 2H), 6.57 (d, 1H), 6.57 (d, 1H) 5.30 (s, 4H).

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 194.9, 148.2, 134.6, 133.3, 119.9, 118.6, 109.7.

LC / MS: Anal. Calcd. ^{For [M + H] + C} 13 H 10 N 2 O: 211.0866; found 211.0867.

< Example  1> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((3,3'- Dimethyl - [1,1'- Biphenyl ] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (9.47 g, 44.0 mmol), EDC (9.97 g, 52.0 mmol), and ortho-tolidine (4.25 g, 20.0 mmol) in a mixture at room temperature and CH ₂ Cl ₂ (30 mL) And stirred for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, 2S, 2'S) -dio-tert-butyl 2,2 '- (((3,3'-dimethyl- [1,1'- biphenyl] -4,4'-diyl) bis (azodioyl) ) Bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (11.3 g, 93%).

The compound (144 mg, 0.238 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (1 mL) and CH ₂ Cl ₂ (1 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure, and a solution of i- Pr ₂ NEt (208 μL, 1.192 mmol) dissolved in CH ₂ Cl ₂ (1 mL) was added over 4 minutes, and further EDC (119 mg, 0.620 mmol) The reaction mixture of the compound (100 mg, 0.572 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (77 mg, 41%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.31 (s, 2H), 7.74 (d, 2H), 7.54-7.23 (m, 16H), 5.52 (d, 2H), 4.52 ( m, 2H), 3.85 (m, 2H), 3.52 (s, 6H), 3.18 (m, 2H), 2.28 (s, 6H), 2.00 - 1.82 (m, 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.1, 168.8, 156.1, 137.1, 136.5, 135.4, 132.2, 128.6, 128.2, 128.1, 128.0, 125.2, 123.9, 60.6, 56.8, 51.6, 46.9, 29.1, 24.3, 17.9;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₄ H ₄₈ N ₆ O ₈ : 789.3606; found 789.3597.

< Example  2> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'-bis Trifluoromethyl ) - [1,1'- Biphenyl ] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine Diyl)) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (3.23 g, 15.0 mmol), EDC (3.12 g, 16.3 mmol), and 2,2'-bis (trifluoromethyl) benzidine (2.00 g, 6.3 mmol) to CH ₂ Cl ₂ (20 mL) and stirred at room temperature for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, (2,2'-bis (trifluoromethyl) - [1,1'-biphenyl] -4,4'-diyl) bis (Aminodiacyl) bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (4.3 g, 96%).

The compound (357 mg, 0.5 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (434 μL, 2.5 mmol) dissolved in CH ₂ Cl ₂ (2 mL) was added over 4 min and further EDC (249 mg, 1.3 mmol) The reaction mixture of the compound (251 mg, 1.2 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (145 mg, 32%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.29 (s, 2H), 8.21 (d, 2H), 7.83 (m, 2H), 7.75 (d, 2H), 7.43-7.05 ( (m, 2H), 3.51 (s, 6H), 3.20 (app br d, 2H), 2.06-1.82 8H);

¹³ C NMR (DMSO- d ₆ ,? = 39.52 ppm, 100 MHz): 171.26, 168.75, 156.42, 139.30, 137.16, 132.79, 131.36, 128.89, 128.35, 128.25, 125.07, 122.88, 121.81, 116.37, 61.05, 56.97, 51.87, 47.22, 29.45, 24.51;

¹⁹ F NMR (DMSO-d ₆ , 377 MHz):? - 57.28;

LC / MS: Anal. Calcd. ^{For [M + H] + C} 44 H 42 F 6 N 6 O 8: 897.3041; found 897.3046.

< Example  3> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'- Dimethyl - [1,1'- Biphenyl ] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (2.23 g, 10.4 mmol), EDC (2.35 g, 12.3 mmol), and meta- tolidine (1.0 g, 4.7 mmol) in a mixture at room temperature and CH ₂ Cl ₂ (10 mL) And stirred for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, 2S, 2'S) -di-tert-butyl 2,2 '- (((2,2'-dimethyl- [1,1'- biphenyl] -4,4'-dyl) bis (azodioyl) ) Bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (2.75 g, 96%).

The compound (292 mg, 0.509 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (443 μL, 2.54 mmol) dissolved in CH ₂ Cl ₂ (2 mL) was added over 4 min and further EDC (253 mg, 1.32 mmol) The reaction mixture of the compound (256 mg, 1.22 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (292 mg, 73%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.84 (s, 2H), 7.73 (d, 2H), 7.59 (s, 2H), 7.48-7.14 (m, 12H), 6.87 ( (d, 2H), 5.51 (d, 2H), 4.42 (m, 2H), 3.84 (m, 2H), 3.55 (m, 8 H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.2, 168.4, 156.1, 137.9, 137.1, 135.8, 135.8, 129.58, 128.62, 128.1, 127.9, 120.5, 116.7, 60.7, 56.8, 51.7, 47.0, 29.4, 24.3, 19.8;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₄ H ₄₈ N ₆ O ₈ : 789.3606; found 789.3605.

< Example  4> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- ((9H- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl Tan-2,1- Dai )) Dicarbamate  Produce

N- Boc-L-proline (323 mg, 1.5 mmol), EDC (312 mg, 1.63 mmol) and 2,7-diaminofluorene (123 mg, 0.63 mmol) were dissolved in CH ₂ Cl ₂ And the mixture was stirred at room temperature for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, Bis (carbonyl)) bis (pyrrolidine-1, 2'S) di-tert-butyl 2,2 ' -Carboxylate) as a solid (359 mg, 97%).

The compound (300 mg, 0.51 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (441 μL, 2.5 mmol) dissolved in CH ₂ Cl ₂ (2 mL) was added over 4 min and further EDC (253 mg, 1.3 mmol) The reaction mixture of the compound (255 mg, 1.3 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue, which was applied to flash chromatography (eluent: EtOAc / hexane mixture) to give the target compound as a white solid (198 mg, 50%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.92 (s, 2H), 7.91 (s, 2H), 7.75-7.69 (m, 4H), 7.56 (d, 2H), 7.44- 2H), 2.04-1.78 (m, 2H), 3.51 (m, 2H), 3.53 (m, 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.2, 168.5, 156.2, 143.6, 137.5, 137.2, 136.4, 128.7, 128.1, 127.9, 119.6, 118.1, 116.2, 60.8, 56.8, 51.7, 47.0, 36.7, 29.4, 24.3;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₃ H ₄₄ N ₆ O ₈ : 773.3293; found 773.3296.

< Example  5> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'- Difluoro - [1,1'-biphenyl] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine Diyl)) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (2.4 g, 10.9 mmol), EDC (2.3 g, 11.8 mmol), and 4,4'-diamino biphenyl (1.0 g, 4.5 mmol) with 2,2'-difluoro Were mixed in CH ₂ Cl ₂ (15 mL) and stirred at room temperature for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, 2S, 2'S) -di-tert-butyl 2,2 '- (((2,2'-difluoro- [1,1'-biphenyl] -4,4'- )) Bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (2.6 g, 93%).

The compound (245 mg, 0.42 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure, and a solution of i- Pr ₂ NEt (370 μL, 2.1 mmol) dissolved in CH ₂ Cl ₂ (2 mL) was added over 4 minutes, and further EDC (210 mg, 1.1 mmol) The reaction mixture of the compound (212 mg, 1.0 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (134 mg, 40%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.43 (s, 2H), 7.74-7.71 (m, 3H), 7.46-7.11 (m, 15H), 5.51 (d, 2H), 4.43 (m, 2H), 3.83 (m, 2H), 3.54 (s, 6H), 3.19 (m, 2H), 2.05 - 1.77 (m, 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.8, 168.4, 160.0, 158.0, 156.1, 140.5, 137.2, 131.6, 128.6, 128.5, 128.1, 127.9, 127.6, 117.1, 115.1, 106.3, 106.1, 60.8, 56.7, 51.7, 47.0, 29.3, 24.3;

_{^{19 F NMR (DMSO-d 6}} , 377 MHz,): δ-73.45;

LC / MS: Anal. Calcd. ^{For [M + H] + C} 42 H 42 F 2 N 6 O 8: 797.3105; found 797.3112.

< Example  6> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'- Dichloro - [1,1'-biphenyl] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- die Work)) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (2.4 g, 10.9 mmol), EDC (2.3 g, 11.8 mmol), and 4,4'-diamino-2,2'-dichloro-biphenyl (1.15 g, 4.5 mmol) to Were mixed in CH ₂ Cl ₂ (15 mL) and stirred at room temperature for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, 2S, 2'S) -di-tert-butyl 2,2 '- (((2,2'-dichloro- [1,1'- biphenyl] -4,4'- ) Bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (2.8 g, 95%).

The compound (245 mg, 0.4 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (4 mL) and CH ₂ Cl ₂ (4 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (347 μL, 2.0 mmol) dissolved in CH ₂ Cl ₂ (4 mL) was added over 4 min and EDC (199 mg, 1.04 mmol) The reaction mixture of the compound prepared in Preparation Example 1 (200 mg, 0.96 mmol) was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (146 mg, 44%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.14 (s, 2H), 7.97-7.91 (m, 2H), 7.75 (d, 2H), 7.65-7.55 (m, 2H), (M, 2H), 3.39 (m, 2H), 3.50 (m, 2H) 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.8, 168.5, 156.2, 139.9, 137.1, 132.6, 132.1, 131.7, 128.6, 128.1, 127.9, 119.3, 117.7, 60.8, 56.73, 51.68, 47.0, 29.3, 24.3;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₂ H ₄₂ Cl ₂ N ₆ O ₈ : 829.2514; found 829.2518.

< Example  7> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'- Dive Lomo - [1,1'-biphenyl] -4,4'- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- die Work)) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-L- proline (755 mg, 3.5 mmol), EDC (729 mg, 3.8 mmol), and 4,4'-diamino-2,2'-dibromo-biphenyl (500 mg, 1.46 mmol) to Were mixed in CH ₂ Cl ₂ (5 mL) and stirred at room temperature for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, 2S, 2'S) -di-tert-butyl 2,2 '- (((2,2'-dibromo- [1,1'- biphenyl] -4,4'-diyl) bis (azodioyl) ) Bis (carbonyl)) bis (pyrrolidine-1-carboxylate) as a solid (991 mg, 92%).

The compound (260 mg, 0.353 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (3078 μL, 1.77 mmol) dissolved in CH ₂ Cl ₂ (4 mL) was added over 4 min and further EDC (176 mg, 0.92 mmol) The reaction mixture of the compound (177 mg, 0.85 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (110 mg, 42%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.13 (s, 2H), 8.32-8.12 (m, 2H), 7.92-7.61 (m, 4H), 7.41-7.13 (m, 12H 2H), 2.05-1.81 (m, 2H), 3.51 (m, 2H), 5.51 (d, 2H), 4.39 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.7, 168.5, 156.2, 139.8, 137.1, 135.9, 131.3, 128.6, 128.1, 127.9, 123.0, 122.3, 118.1, 60.8, 56.7, 51.7, 47.0, 29.3, 24.3;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₂ H ₄₂ Br ₂ N ₆ O ₈ : 917.1504; found 917.1521.

< Example  8> Dimethyl  ((1R, 1'R) - ((2R, 2'R) -2,2 '- (([ Biphenyl ] -4,4'-diylbis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N- Boc-D-proline (771 mg, 3.6 mmol), EDC (812 mg, 4.2 mmol) and benzidine (300 mg, 1.63 mmol) were mixed in CH ₂ Cl ₂ (4 mL) Respectively.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, Bis (carbonyl)) bis (2,2 ', 2'-di-tert-butyl 2,2' (Pyrrolidine-1-carboxylate) as a solid (930 ng, 99%).

The compound (300 mg, 0.52 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure, and a solution of i- Pr ₂ NEt (455 μL, 2.6 mmol) dissolved in CH ₂ Cl ₂ (3 mL) was added over 4 minutes, and further EDC (258 mg, 0.620 mmol) The reaction mixture of the compound (260 mg, 1.24 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (171 mg, 57%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.14 (s, 2H), 7.68-7.60 (m, 9H), 7.46-7.30 (m, 11H), 5.49 (d, 2H), 4.53 (m, 2H), 3.68 (m, 2H), 3.54 (s, 6H), 3.12 (m, 2H), 2.00 - 2.13 (m, 2H), 1.89 - 1.82 (m, 6H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.2, 168.1, 156.4, 138.2, 136.9, 134.4, 128.4, 128.4, 127.8, 126.4, 119.4, 60.6, 56.6, 51.6, 46.9, 29.4, 24.7;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₂ H ₄₄ N ₆ O ₈ : 761.3293; found 761.3281.

< Example  9> Dimethyl  ((1R, 1'R) - ((5S, 5'S) -5,5 '- (([ Biphenyl ] -4,4'-diylbis ( Azandai I )) Bis ( Carbonyl )) Bis (3- Oxopyrrolidine -5,1- Dai )) Bis (2-oxo-1- Neil Tan-2,1- Dai )) Dicarbamate  Produce

(33 mg, 1.5 mmol), EDC (303 mg, 1.6 mmol), and benzidine (112 mg, 0.61 mmol) were mixed in CH ₂ Cl ₂ (2 mL) and treated with N- Boc-4-oxo-L- Lt; / RTI &gt; for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with 1N hydrochloric acid aqueous solution and brine, dried over magnesium sulfate, filtered, concentrated under reduced pressure, Bis (carbonyl)) bis (3S, 5S, 5'S) -di-tert-butyl 5,5 ' -Oxopyrrolidine-1-carboxylate) as a solid (320 mg, 87%).

The compound (161 mg, 0.27 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (2 mL) and CH ₂ Cl ₂ (2 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure and a solution of i- Pr ₂ NEt (232 μL, 1.3 mmol) dissolved in CH ₂ Cl ₂ (2 mL) was added over 4 min and further EDC (132 mg, 0.69 mmol) The reaction mixture of the compound (133 mg, 0.64 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (48 mg, 23%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.38 (s, 2H), 7.92 (d, 2H), 7.70-7.10 (m, 18H), 5.46 (d, 2H), 4.94 ( d, 2H), 4.25 (d, 2H), 3.89 (d, 2H), 3.54 (s, 6H), 3.06 (m, 2H), 2.54 (m, 2H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 208.5, 170.1, 169.4, 156.1, 137.8, 136.8, 134.7, 128.6, 128.2, 128.1, 126.5, 119.7, 57.4, 56.9, 53.1, 51.7, 40.4;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₂ H ₄₀ N ₆ O ₁₀ : 789.2879; found 789.2877.

< Example  10> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (([ Biphenyl ] -4,4'-diylbis ( Azandai I )) Bis ( Carbonyl )) Bis (Piperidin-2,1- Dai )) Bis (2-oxo-1- Pe Ethyl-2,1- Dai )) Dicarbamate  Produce

N -Boc-L- mixed at l cholic acid (400 mg, 1.75 mmol), EDC (363 mg, 1.9 mmol), and benzidine (134 mg, 0.73 mmol) to CH ₂ Cl ₂ (7 mL) and 2 hours at room temperature Lt; / RTI &gt;

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with a 1N aqueous hydrochloric acid solution and brine, dried over magnesium sulfate, filtered and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to give (2S, 2'S) -di-tert- butyl 2,2 ' (186 mg, 42%) was obtained as a solid. MS (m / z) &lt;'&gt;

The compound (73 mg, 0.12 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (1 mL) and CH ₂ Cl ₂ (1 mL), and the mixture was stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure, and a solution of i- Pr ₂ NEt (105 μl, 0.60 mmol) dissolved in DMF (1 mL) was added over 4 minutes, and further EDC (60 mg, 0.31 mmol) (60 mg, 0.29 mmol) was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the desired compound as a solid (12 mg, 13%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.99-9.84 (s, 2H), 7.85-7.30 (m, 20H), 5.73-5.64 (m, 2H), 5.17 / 4.85 (m 2H), 1.76 (m, 2H), 1.63-1.24 (m, 2H), 4.45 , 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.0, 169.3, 168.5, 156.2, 138.0, 137.2, 134.5, 128.6, 128.4, 128.2, 127.7, 126.5, 120.2, 119.7, 67.0, 55.5, 52.8, 51.6, 43.2, 27.5, 25.1, 24.6, 19.7;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₄ H ₄₈ N ₆ O ₈ : 789.3606; found 789.3605.

< Example  11> Dimethyl  ((1R, 1'R) - ((2R, 2'R) -2,2 '- (([ Biphenyl ] -4,4'-diylbis ( Azandai I )) Bis ( Carbonyl )) Bis (Piperidin-2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

N -Boc-D- l cholic acid (500 mg, 2.2 mmol), mix in EDC (452 mg, 2.4 mmol) , and benzidine (167 mg, 0.91 mmol) to CH ₂ Cl ₂ (4 mL) and 2 hours at room temperature Lt; / RTI &gt;

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with a 1N aqueous hydrochloric acid solution and brine, dried over magnesium sulfate, filtered and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain (2R, 2'R) -di-tert- butyl 2,2 ' (190 mg, 35%) was obtained as a solid. MS (m / z) &lt; RTI ID = 0.0 & ).

The compound (122 mg, 0.2 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (1 mL) and CH ₂ Cl ₂ (1 mL), and the mixture was stirred at room temperature for 5 hours. Volatile components were removed under reduced pressure, and a solution of i- Pr ₂ NEt (195 μL, 1.0 mmol) dissolved in DMF (1 mL) was added over 4 minutes, and further EDC (100 mg, 0.52 mmol) (101 mg, 0.48 mmol) was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and subjected to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (23 mg, 15%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.98-9.84 (s, 2H), 7.84-7.28 (m, 20H), 5.72-5.64 (m, 2H), 5.16 / 4.85 (m 2H), 1.76 (m, 2H), 1.63-1.23 (m, 2H), 4.45 (m, 2H), 3.55 , 8H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.0, 169.3, 168.5, 156.2, 138.0, 137.2, 134.5, 128.6, 128.4, 128.2, 127.7, 126.5, 120.2, 119.7, 67.0, 55.5, 52.8, 51.6, 43.2, 27.5, 25.1, 24.6, 19.7;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₄ H ₄₈ N ₆ O ₈ : 789.3606; found 789.3605.

< Example  12> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (([ Biphenyl ] -4,4'-diylbis ( Azandai I )) Bis ( Carbonyl )) Bis (2- Methylpyrrolidine -2,1- Dai )) Bis (2-oxo-1- Neil Tan-2,1- Dai )) Dicarbamate  Produce

-Boc-N-methyl-a- -L- proline (200 mg, 0.87 mmol), EDC (181 mg, 0.95 mmol), and benzidine (67 mg, 0.36 mmol) and the mixture at room temperature in CH ₂ Cl _{2 (2} mL) Lt; / RTI &gt; for 2 hours.

Next, the compound obtained above was fractionated into CH ₂ Cl ₂ and H ₂ O, and the organic layer was washed with a 1N aqueous hydrochloric acid solution and brine, dried over magnesium sulfate, filtered and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to give (2S, 2'S) -di-tert- butyl 2,2 ' (2-methylpyrrolidine-1-carboxylate) as a solid (86 mg, 31 &lt; RTI ID = 0.0 & %).

The compound (144 mg, 0.238 mmol) obtained above was placed in a mixed solvent of CF ₃ CO ₂ H (1 mL) and CH ₂ Cl ₂ (1 mL) and stirred at room temperature for 5 hours. The volatiles were removed under reduced pressure, and a solution of i- Pr ₂ NEt (208 μL, 1.192 mmol) dissolved in CH ₂ Cl ₂ (1 mL) was added over 4 minutes, and further EDC (119 mg, 0.620 mmol) The reaction mixture of the compound (100 mg, 0.572 mmol) prepared in Preparation Example 1 was stirred at room temperature for 75 minutes. Next, the reaction residue was fractionated into CH ₂ Cl ₂ and H ₂ O, and the obtained organic layer was washed with H ₂ O and brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. A silica gel mesh was prepared from the residue and applied to flash chromatography (eluent: EtOAc / hexane mixture) to obtain the target compound as a white solid (77 mg, 41%).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.03 (s, 1H), 8.89 (s, 1H), 7.77-7.57 (m, 10H), 7.40-7.32 (m, 10H), 3H), 3.48 (m, 1H), 3.21 (m, 1H), 2.16 (m, -2.08 (m, 2H), 1.91-1.80 (m, 6H), 1.55 (s, 3H), 1.43 (s, 3H);

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 171.8, 171.7, 168.2, 167.7, 156.4, 156.2, 138.2, 138.0, 137.2, 136.5, 134.7, 134.3, 128.7, 128.4, 128.22, 128.17, 127.70, 127.68, 126.09, 126.05, 120.9, 120.3, 67.6, 67.5, 57.2, 57.0, 51.7, 51.6, 47.7, 47.5, 23.5, 23.1, 20.6, 20.5;

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₄ H ₄₈ N ₆ O ₈ : 789.3606; found 789.3600.

< Example  13> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9,9- Difluoro -9H- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dill )) Dicarbamate  Produce

Step 1: (2S, 2 ' S) - die - tert - butyl 2,2 '- (((9,9- Difluoro -9H- Fluorene -2,7-diyl) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -One- Carboxylate )

CH ₂ Cl ₂ (1 mL) of N -Boc-L-proline (24 mg, 0.11 mmol) was dissolved in, EDC (24 mg, 0.12 mmol ) and 9,9-difluoro-a obtained in Preparation Example 3-9 The mixture of H -fluorene-2,7-diamine (11 mg, 0.047 mmol) was mixed at ambient temperature for 2 hours and then layered using CH ₂ Cl ₂ and H ₂ O. The organic layer was washed with 1 N aq HCl solution and brine, dried over MgSO ₄ , filtered, and concentrated in vacuo. Without purification, the desired product (28 mg, 95%) was obtained in solid form.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.29 (app br s, 2H), 8.06 (s, 1H), 8.02 (s, 1H), 7.69-7.66 (m, 4H), (M, 2H), 2.23-2.16 (m, 2H), 1.92-1.80 (m, 2H), 3.45-3. , 6H), 1.40 (app br s, 9H), 1.27 (app br s, 9H).

_{^{13 C NMR (DMSO-d 6}} , δ = 39.52 ppm, 100 MHz): 174.3, 173.9, 171.9, 171.5, 153.6, 153.1, 139.4, 133.56, 133.49, 122.8, 122.7, 121.2, 114.5, 114.4, 78.8, 78.6, 60.5, 59.3, 58.6, 46.6, 46.2, 46.1, 31.0, 30.3, 28.2, 28.0, 27.9, 27.7, 24.0, 23.4.

_{^{19 F NMR (DMSO-d 6}} , 377 MHz,): δ-108.9, -109.0.

LC / MS: Anal. Calcd. ^{For [M + H] + C} 33 H 40 F 2 N 4 O 6: 627.2989; found 627.2997.

Step 2: Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9,9- Difluoro -9H- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dill )) Dicarbamate  Produce

_{CF 3 CO 2 H (1 mL} ) and CH ₂ Cl ₂ (1 mL) of (2S, 2'S) obtained in Step 1 was dissolved in -di -tert- butyl 2,2 '- (((9,9-dimethyl Bis (pyrrolidine-1-carboxylate) (27 mg, 0.043 mmol) was added to a solution of 5 &lt; RTI ID = 0.0 &gt;Lt; / RTI &gt; The volatiles (volatile component) is removed in a vacuum, dissolved in EDC (21 mg, 0.11 mmol) , Cap (22 mg, 0.10 mmol) for one batch _{(batches) CH 2 Cl 2 (} 1 mL) for 4 min. i- Pr ₂ NEt (28 mL, 0.22 mmol), and the reaction mixture was mixed at room temperature for 75 minutes. After layer separation using CH ₂ Cl ₂ and H ₂ O, the organic layer was washed with H ₂ O and brine, dried over MgSO ₄ , filtered and concentrated in vacuo. A silica gel mesh was prepared from the residue and the desired product (18 mg, 52%) was obtained as a solid via flash chromatography (silica gel: EtOAc / hexane as eluent).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.14 (s, 2H), 8.05 (s, 2H), 7.77 (d, 2H), 7.71 (s, 4H), 7.43-7.10 ( (m, 2H), 5.51 (d, 2H), 4.39 (m, 2H), 3.85 (m, 2H), 3.55 (s, 6H), 3.21 (m, 2H), 2.06-1.

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.6, 168.5, 156.2, 139.3, 137.3, 137.0, 128.6, 128.4, 128.1, 127.9, 127.6, 122.8, 121.2, 114.5, 60.8, 56.7, 51.6, 47.0, 29.3, 24.3.

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₃ H ₄₂ F ₂ N ₆ O ₈ : 809.3105; found 809.3109.

< Example  14> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9,9- Dimethyl -9H- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

Step 1: (2S, 2 ' S) - die - tert - butyl 2,2 '- (((9,9- Dimethyl -9H- Fluorene -2,7-diyl) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -One- Carboxylate )

CH ₂ Cl ₂ (1 mL) of N -Boc-L-proline (173 mg, 0.80 mmol), EDC (167 mg, 0.87 mmol) and 9,9-dimethyl -9 obtained in Preparation Example 2 dissolved in H - The mixture of fluorene-2,7-diamine (75 mg, 0.33 mmol) was mixed at ambient temperature for 2 hours and then layered using CH ₂ Cl ₂ and H ₂ O. The organic layer was dried with 1 N aq HCl and washed with brine and MgSO _4, filter, and concentrated in vacuo. Without further purification, the desired product (201 mg, 97%) was obtained in solid form.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.10 (app br s, 2H), 7.84 (s, 1H), 7.79 (s, 1H), 7.66 (d, 2H), 7.51 ( 2H), 4.31-4.28 (m, 1H), 4.24-4.21 (m, 1H), 3.44-3.40 (m, 2H) 1.95-1.80 (m, 6H), 1.40 (app br s, 9H), 1.34 (s, 6H), 1.28 (app br s, 9H).

_{^{13 C NMR (DMSO-d 6}} , δ = 39.52 ppm, 100 MHz): 174.3, 173.9, 171.4, 171.0, 153.6, 153.2, 138.1, 138.0, 133.7, 133.6, 119.7, 118.4, 118.2, 113.8, 113.6, 78.6, 78.5, 60.4, 60.0, 46.6, 46.1, 42.2, 42.1, 31.0, 30.3, 28.2, 28.1, 27.9, 27.1, 24.0, 23.4.

LC / MS: Anal. Calcd. ^{For [M + H] + C} 35 H 42 N 4 O 6: 619.3490; found 619.3496.

Step 2: Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9,9- Dimethyl -9H- Fluorene -2,7-diyl) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

(2S, 2'S) -di-tert-butyl 2,2 '- (((9,9-difluoro-9H- fluoren- (2S, 2'S) -di-tert-butyl 2,2 '- (((9,9-dimethyl- (Pyrrolidine-1-carboxylate) (200 mg, 0.32 mmol) was used in place of the bis (cyclopentadienyl) The target product (135 mg, 53%) was obtained in the form of a solid by the same procedure as in the step 13 of Example 13.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 9.96 (s, 2H), 7.86 (s, 2H), 7.74 (d, 2H), 7.69 (d, 2H), 7.51 (dd, 2H), 3.53 (s, 6H), 3.20 (m, 2H), 2.05-1.78 (m, 2H) (m, 8 H), 1.41 (s, 6 H).

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 170.1, 168.3, 156.1, 153.7, 138.0, 137.2, 133.7, 128.6, 128.1, 127.8, 119.8, 118.3, 113.7, 60.7, 56.7, 51.6, 47.0, 46.5, 29.3, 27.2, 24.3.

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₅ H ₄₈ N ₆ O ₈ : 801.3606; found 801.3611.

< Example  15> Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9- Fluorene -2,7-diyl) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Phenyl ethane -2,1- Dai )) Dicarbamate  Produce

Step 1: (2S, 2 ' S) - die - tert - butyl 2,2 '- (((9-oxo-9H- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -One- Carboxylate )

Dissolved in _{CH 2 Cl 2 (1 mL)} N -Boc-L-proline (123 mg, 0.57 mmol), EDC (119 mg, 0.62 mmol) and 2,7-diamino -9 H obtained in Preparation Example 4 -Fluoren-9-one (50 mg, 0.24 mmol) were mixed at ambient temperature for 2 hours and then layered using CH ₂ Cl ₂ and H ₂ O. The organic layer was washed with a 1 N aq HCl solution and brine, and dried, the filter with MgSO _4, and concentrated in vacuo. Without further purification, the desired product (131 mg, 91%) was obtained in the form of a solid.

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.24 (s, 2H), 7.91 (m, 2H), 7.91 (m, 2H), 7.62 (m, 2H), 4.26-4.16 ( (m, 2H), 3.46-3.40 (m, 2H), 3.37-3.31 (m, 2H), 2.24-2.16 1.27 (app br s, 9H).

_{^{13 C NMR (DMSO-d 6}} , δ = 39.52 ppm, 100 MHz): 192.9, 171.9, 171.4, 153.6, 153.1, 139.6, 138.8, 134.2, 125.0, 121.1, 115.0, 78.8, 78.6, 60.5, 60.1, 46.6, 46.1, 31.0, 30.2, 28.2, 28.0, 24.0, 23.4.

LC / MS: Anal. Calcd. ^{For [M + H] + C} 33 H 40 N 4 O 7: 605.2970; found 605.2980.

Step 2: Dimethyl  ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((9- Fluorene -2,7- Dai ) Bis ( Azandai I )) Bis ( Carbonyl )) Bis ( Pyrrolidine -2,1- Dai )) Bis (2-oxo-1- Pe Ethyl-2,1- Dai )) Dicarbamate  Produce

(2S, 2'S) -di-tert-butyl 2,2 '- (((9,9-difluoro-9H- fluoren- (2S, 2'S) -di-tert-butyl 2,2 '- (((9-oxo-9H-fluoro- (Pyrrolidine-1-carboxylate) (14 mg, 0.023 mmol) was used in place of the bis (diphenylphosphino) Step 13 The title compound (12 mg, 66%) was obtained as a solid in the same manner as in step 2).

_{^{1 H NMR (DMSO- d 6,}} δ = 2.5 ppm, 400 MHz): 10.10 (s, 2H), 7.91 (s, 2H), 7.76 (t, 4H), 7.64 (d, 2H), 7.43-7.10 ( (m, 2H), 5.51 (d, 2H), 4.39 (m, 2H), 3.85 (m, 2H), 3.55 (s, 6H), 3.20 (m, 2H), 2.05-1.

_{^{13 C NMR (DMSO- d 6,}} δ = 39.52 ppm, 100 MHz): 192.8, 170.6, 168.5, 156.2, 139.5, 138.8, 137.1, 134.2, 128.6, 128.1, 127.9, 125.0, 121.1, 115.0, 60.8, 56.7, 51.7, 47.0, 29.3, 24.3.

LC / MS: Anal. Calcd. For [M + H] ⁺ C ₄₃ H ₄₂ N ₆ O ₉ : 787.3086; found 787.3089.

Hereinafter, the chemical structural formulas of the compounds prepared in Examples 1-15 are shown in Table 1 below.

Example Chemical structural formula Example Chemical structural formula One

9

2

10

3

11

4

12

5

13

6

14

7

15

8

< Experimental Example  1> Evaluation of hepatitis C virus activity 1

The following experiments were conducted to evaluate the anti-hepatitis C virus activity of the compounds of formula (I) according to the present invention.

Huh7.5.1 cells, a liver cancer cell line, were dispensed into a 12-well cell culture plate at about 50,000 per well. Dissociated cells were supplemented with 10% (v / v) FBS (Fetal Bovine Serum SH30406.02, Hyclone Co.) and 1% (v / v) antibiotics (penicillin / streptomycin solution SV30010, Hyclone Co.) The cells were cultured for 24 hours in a 6.0% carbon dioxide cell incubator (CO ₂ Incubator 311, Forma Scientific Co, Lnc. USA) at 37 ° C using a DMEM culture medium (Ducos-modified Eagle culture solution 12800-017, GIBCO Co.) And attached to the floor. Subsequently, hepatitis C virus (JFH-5aFlucm4, PLoS One. 2011; 6 (8): e228808.) With the Renilla luciferase gene was inoculated into the cells for 3 hours as a reporter Respectively. After 3 hours of virus inoculation, the cell culture broth was removed and a new cell culture broth prepared by adding the compounds prepared in Examples 1 to 15 according to the present invention to the culture broth at a concentration of 1 [mu] M was injected into each well, And incubated for 3 days in a 6.0% carbon dioxide cell incubator (CO ₂ Incubator 311, Forma Scientific Co, Lnc. USA) at 37 ° C. Three days after the infection, the cell culture medium was collected separately to measure the cytotoxicity of the following <Experimental Example 3>. Cells attached to the plate were washed with phosphate buffered saline (PBS) and lysed by treatment with 100 μl of 1 × indirect lysis buffer (passive lysis buffer, promega, E1941). The lysed cell lysate (10 ㎕) was injected into the Renilla luciferase assay reagent (50 ㎕) of the Renilla Luciferase assay system (Promega, E2820) and mixed to obtain a luminometer (GLOMAX 20/20 Luminometer, Promega) using luciferase measure program for 10 seconds. At this time, the luminescence quantities of the treated and untreated groups treated with the compounds of Examples 1 to 15 according to the present invention were measured three times, and their average was compared and analyzed. The luminescence amount according to the concentration of each compound was measured with a sigma plot The maximum maximal effective concentration (EC ₅₀ ) was calculated using a program (sigma plot program) and is shown in Table 2 below.

Example EC ₅₀ Rating One C 2 A 3 A 4 B 5 A 6 A 7 A 8 C 9 A 10 C 11 C 12 C 13 A 14 A 15 B

In Table 2,

Grade A is less than EC ₅₀ 1 nM;

B grade is EC ₅₀ 1 nM - 100 nM;

Class C indicates that the EC _{50 is} more than 100 nM.

As shown in Table 2, the compounds according to the present invention have antiviral effects against hepatitis C virus present in liver cancer cell lines. In particular, in Examples 2, 3, 5, 6, 7, 9, 13 and 14, the maximum effective concentration (EC ₅₀ ) was less than 1 nM at 50%

Therefore, the compounds according to the present invention are excellent in anti-C hepatitis virus effect against hepatitis C virus, so that they can be used for the treatment of acute hepatitis C, chronic hepatitis C, liver cirrhosis or hepatocellular carcinoma And can be usefully used as a pharmaceutical composition for preventing or treating the same liver disease.

< Experimental Example  2> Evaluation of anti-hepatitis C virus activity 2

The following experiments were carried out in order to evaluate the anti-hepatitis C virus activity of the compound of formula (1) according to the present invention, i.e., the activity of the reproductive virus.

About 50,000 huh7.5.1 cells with NK / R2AN replicon, which can measure the replication and translation process of hepatitis C virus, were dispensed into 12-well cell culture plates. Divided cells were treated with antibiotics (penicillin / streptomycin solution SV30010, Hyclone Co.) at a final concentration of 10% (v / v) FBS (Fetal Bovine Serum SH30406.02, Hyclone Co.) (CO ₂ Incubator 311, manufactured by Forma Scientific Co, Inc) at 37 ° C using a DMEM culture medium (Ducose-modified eagle culture solution 12800-017, GIBCO Co.) supplemented with G418 (600 μg / mL, Calbiochem) Lnc. USA) for 24 hours to attach the cells to the bottom of the plate. Then, the cell culture medium in which the cells were grown was removed, and a new cell culture medium prepared by adding the compounds prepared in Examples 1 to 15 according to the present invention to the culture medium at a concentration of 40 pM to 1 μM, respectively, was injected into each well , And incubated for 3 days in a 6.0% carbon dioxide cell incubator (CO ₂ Incubator 311, Forma Scientific Co, Lnc. USA) at 37 ° C. Three days after the infection, the cell culture medium was collected separately to measure the cytotoxicity of the following <Experimental Example 3>. Cells attached to the plate were washed with phosphate buffered saline (PBS) and treated with 100 μl of 1 × indirect lysis buffer (E1941). The lysed cell lysate (10 ㎕) was injected into the Renilla luciferase assay reagent (50 ㎕) of the Renilla Luciferase assay system (Promega, E2820) and mixed to obtain a luminometer (GLOMAX 20/20 Luminometer, Promega) using luciferase measure program for 10 seconds. At this time, the luminescence quantities of the treated and untreated groups treated with the compounds of Examples 1 to 15 according to the present invention were measured three times, and their average was compared and analyzed. The luminescence amount according to the concentration of each compound was measured with a sigma plot The maximum maximal effective concentration (EC ₅₀ ) was calculated using a program (sigma plot program) and is shown in Table 3 below.

Example EC ₅₀ Rating One C 2 A 3 A 4 A 5 A 6 A 7 A 8 C 9 A 10 C 11 D 12 C 13 A 14 A 15 A

In Table 3,

Grade A is less than EC ₅₀ 100 pM;

Class B is EC ₅₀ 100 pM - 1 nM;

Class C is EC ₅₀ 1 nM - 100 nM;

The grade D indicates that the EC _{50 is} more than 100 nM.

As shown in Table 3 above, the compounds of the examples according to the present invention were found to have an excellent antiviral effect against hepatitis C virus. More specifically, the compounds prepared in Examples 2, 3, 4, 5, 6, 7, 9, 13, 14 and 15 had an EC ₅₀ value of less than 100 pM and showed antiviral activity against hepatitis C virus It was confirmed to be excellent.

Therefore, the compounds according to the present invention are highly effective against the hepatitis C virus even at a relatively low concentration against the hepatitis C virus, so that the acute hepatitis C, chronic hepatitis C, cirrhosis Or liver diseases such as hepatocellular carcinoma.

< Experimental Example  3> Assessment of cytotoxicity

In order to evaluate the cytotoxicity of the compounds of the present invention represented by formula (1) according to the present invention, the following experiment was conducted.

20 μl of the culture medium per well was used according to the cytotoxicity assay kit (Lonza, LT07-117), and AK detection reagent (ToxiLight Non-destructive Cytotoxicity Bio Assay Kit LT07-117, Lonza Co., was added to a 100 ㎕) was allowed to stand for 5 minutes Victor _{^{3 (VICTOR 3 TM wallaac 1420-051 Multiabel}} plate Counter, PerkinElmer Inc. Boston, MA, USA) Wallac 1420 workstation (Wallac 1420 workstation to) program To measure the degree of light emission for 1 second at a wavelength of 565 nm. The light emission amounts of the compound treatment group and the non-treatment group of Examples 1 to 12 according to the present invention were compared and analyzed. As a result, it was confirmed that when treated at a concentration of 1 μM, there was no cytotoxicity, and when treated at a concentration of 25 μM, the fluorescence of the kit for confirming cytotoxicity was confirmed to be emitted at 565 nm, indicating cytotoxicity. From this, it can be seen that the compound represented by the formula (1) according to the present invention has no side effects due to cytotoxicity.

< Formulation example  1> Sanje  Produce

The compound of formula (1) 2 g

Lactose 1 g

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

< Formulation example  2> Preparation of tablets

The compound of formula (1) 100

Corn starch 100

Lactose 100

Magnesium stearate 2

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

< Formulation example  3> Preparation of capsules

The compound of formula (1) 100

Corn starch 100

Lactose 100

Magnesium stearate 2

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

< Formulation example  4> Preparation of injection

The compound of formula (1) 100

Mannitol 180

Na ₂ HPO ₄ 2H ₂ O 26

Distilled water 2974

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

< Formulation example  5> Manufacture of health functional foods

The compound of formula (1) 1000

Vitamin mixture Suitable amount

Vitamin A acetate 70

Vitamin E 1.0

vitamin 0.13

Vitamin B2 0.15

Vitamin B6 0.5

Vitamin B12 0.2

Vitamin C 10

Biotin 10

Nicotinic acid amide 1.7

Folic acid 50

Calcium pantothenate 0.5

Mineral mixture Suitable amount

Ferrous sulfate 1.75

Zinc oxide 0.82

Magnesium carbonate 25.3

Potassium monophosphate 15

Dicalcium phosphate 55

Potassium citrate 90

Calcium carbonate 100

Magnesium chloride 24.8

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 modified, and the above components may be mixed , Granules may be prepared and used in the manufacture of a health functional food composition according to a conventional method.

< Formulation example  6> Manufacture of health drinks

The compound of formula (1) 1000

Citric acid 1000

oligosaccharide 100 g

Plum concentrate 2 g

Taurine 1 g

Purified water is added to the entire 900

The above ingredients were mixed according to the usual health drink manufacturing method, and the mixture was stirred and heated at 85 for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, And used for manufacturing.

Although the composition ratio is relatively mixed with the ingredient suitable for the favorite drink, it is also possible to arbitrarily modify the blending ratio according to the regional or national preference such as the demand class, demand country, use purpose, and the like.

< Formulation example  7> Manufacture of other health functional foods

7-1. Manufacturing of beverages

honey 522

Dioctanoic acid amide 5

Nicotinic acid amide 10

Sodium riboflavin hydrochloride 3

Pyridoxine hydrochloride 2

Inositol 30

Ortho acid 50

The compound of formula (1) 0.48-1.28

water 200

A beverage was prepared using the above-mentioned composition and content by a conventional method.

7-2. Of chewing gum  Produce

Gum base 20%

Sugar 76.36-76.76%

The compound of formula (1) 0.24-0.64%

Fruits One %

water 2 %

Chewing gum was prepared using the above-mentioned composition and content by a conventional method.

7-3. Manufacture of candy

Sugar 50-60%

corn syrup 39.26-49.66%

The compound of formula (1) 0.24-0.64%

Orange incense 0.1%

The composition and the content of the candy were prepared using a conventional method.

7-4. Manufacture of flour food products

0.5 to 5 parts by weight of the benzidine derivative of the formula (1) was added to 100 parts by weight of wheat flour, and bread, cake, cookies, crackers and noodles were prepared by using this mixture.

7-5. dairy product( dairy products )

5 to 10 parts by weight of the benzidine derivative of the formula (1) was added to 100 parts by weight of milk, and various dairy products such as butter and ice cream were prepared using the milk.

7-6. Solar  Produce

Brown rice 30%

Yulmu 15%

barley 20%

Perilla 7%

Black beans 7%

Black sesame 7%

The compound of formula (1) 3%

wisdom 0.5%

Rehab 0.5%

Brown rice, barley, glutinous rice, and yulmu were alphalized by a known method and dried, and the powder was prepared into a powder having a particle size of 60 mesh by a pulverizer. Black beans, black sesame seeds, and perilla seeds were steamed and dried by a known method, and then powdered with a particle size of 60 meshes by a grinder. The grains and seeds pulverized by the powder were mixed with the benzidine derivative of the formula (I) of the present invention in the same ratio as above.

Claims (10)

Claims 1. A compound represented by the following formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
R ¹ and R ² are independently -H, -OH, a halogen, C _{1 -10} straight or branched chain alkyl, C _{1 -10} linear or branched alkoxy, unsubstituted or substituted C _{6 -10} of the aryl, or - NHC (= O) R &lt; ¹² &gt;
Here, the above substituted aryl of _{6 -10} C is one or more substituents selected from the group consisting of straight or branched alkyl, straight-chain or branched alkoxy and halogen of C _{1 -5} C for _{1 -5} may be substituted,
In addition, the R ¹² is -H, or a linear or branched alkoxy of ₁ C and _-5;

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently selected from -H, halogen, unsubstituted or substituted C _{1 -5} straight or branched ego,
Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring of _{5 -6} C with the carbon atoms to which they are connected, the ring include halogen, C ₁ of the C _{5 -6 -5} straight or branched chain alkyl and = O may be substituted;

R ¹¹ is -H, -OH, halogen, straight or branched chain alkoxy of C _{1 -10} straight or branched chain alkyl, C _{1 -10} of, or is = O;

Is a single or double bond;

and a is an integer of 0-3).
The method according to claim 1,
R ¹ and R ² are independently -H, -OH, a halogen, C _{1 -5} straight or branched chain alkyl, C _{1 -5} straight or branched chain alkyl, unsubstituted or substituted C _{6 -8} in the aryl, or - NHC (= O) R &lt; ¹² &gt;
Wherein said substituted C _{6 -8} aryl may be substituted with one or more substituents selected from the group consisting of C _{1 -3} straight or branched alkyl, C _{1 -3} straight or branched alkoxy and halogen,
Also, R ¹² is -H or C _{1 -3} straight or branched alkoxy;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently selected from -H, halogen, unsubstituted or substituted C _{1 -3} straight or branched ego,
Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring of _{5 -6} C with the carbon atoms to which they are connected, the ring include halogen, C ₁ of the C _{5 -6 -3} straight or branched chain alkyl and = O may be substituted;

R ¹¹ is -H, -OH, halogen, C _{1 -5} straight or branched chain alkyl, C _{1 -5} straight or branched alkoxy, or = O;

Is a single or double bond;

and a is an integer of 0-2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
The method according to claim 1,
R ¹ and R ² are independently phenyl, or methoxycarbonylamino;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently -H, -F, -Cl, -Br, -CF ₃ ,
Here, the R ⁴ and R ^7, or R ⁶ and R ⁹ may form a ring in C ₅ along with the carbon atoms to which they are connected together, with -F, = O, and methyl, the ring of the C ₅ One or more substituents selected from the group consisting of the substituents may be substituted;

R &lt; ¹¹ &gt; is -H, or = O;

Is a single or double bond;

and a is an integer of 0-1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
The method according to claim 1,
The compound represented by the formula (1) is any one selected from the following compound group, its optical isomer, or a pharmaceutically acceptable salt thereof:
(1) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;
(2) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 '- (((2,2'-bis (trifluoromethyl) - [ (2-oxo-1-phenylethan-2-yl) -4,4'-diyl) bis (azodioyl)) bis (carbonyl)) bis (pyrrolidin- 1-yl))) dicarbamate;
(3) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;
(4) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' Carbonyl)) bis (pyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(5) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl)) bis (carbonyl)) bis (pyrrolidine- )) Dicarbamate;
(6) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;
(7) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl) bis (azodiacyl) bis (carbonyl)) bis (pyrrolidine- ) Dicarbamate;
(8) Synthesis of dimethyl ((1R, 1'R) - ((2R, 2'R) -2,2 ' (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate; bis (pyrrolidine-2,1-diyl)) bis
(9) Synthesis of dimethyl ((1R, 1'R) - ((5S, 5'S) -5,5 ' )) Bis (carbonyl)) bis (3-oxopyrrolidin-5,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(10) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (piperidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(11) Synthesis of dimethyl ((1R, 1'R) - ((2R, 2'R) -2,2 ' Bis (carbonyl)) bis (piperidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(12) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (2-methylpyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(13) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' (Azodioyl)) bis (carbonyl)) bis (pyrrolidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate;
(14) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' Bis (carbonyl)) bis (pyrrolidine-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate; And
(15) Synthesis of dimethyl ((1R, 1'R) - ((2S, 2'S) -2,2 ' )) Bis (carbonyl)) bis (pyrrolidin-2,1-diyl)) bis (2-oxo-1-phenylethane-2,1-diyl)) dicarbamate.
As shown in Scheme 1 below,
Reacting a compound represented by formula (2) with a compound represented by formula (3) in an organic solvent to prepare a compound represented by formula (4) (step 1);
Removing the protecting group of the compound represented by the formula (4) prepared in the step (1) to prepare a compound represented by the formula (5) (step 2); And
Reacting the compound represented by the general formula (5) and the compound represented by the general formula (6) to produce the compound represented by the general formula (1) (step 3). Preparation of the indicated compounds:
[Reaction Scheme 1]

(In the above Reaction Scheme 1,
^{R 1, R 2, R 3} , R 4, R 5, R 6, R 7, R 8, R 9, R 10, R 11, a, and

Is as defined in claim 1;
PG is a protecting group).
6. The method of claim 5,
Wherein the organic solvent in Step 1 is at least one selected from the group consisting of methanol, dimethylformamide, tetrahydrofuran, dichloromethane, and toluene.
A pharmaceutical composition for preventing or treating liver disease caused by hepatitis C virus comprising the compound of formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
8. The method of claim 7,
Wherein the liver disease caused by the hepatitis C virus is one selected from the group consisting of acute hepatitis C, chronic hepatitis C, cirrhosis and hepatocellular carcinoma.
A health functional food composition for preventing or ameliorating liver disease caused by hepatitis C virus comprising the compound represented by the general formula (1) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
10. The method of claim 9,
Wherein the liver disease caused by hepatitis C virus is one selected from the group consisting of acute hepatitis C, chronic hepatitis C, cirrhosis and hepatocellular carcinoma.