KR100516432B1 - 2-(4-Substituted-anilino)pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same - Google Patents

Abstract

The present invention relates to a 2- (4-substituted-anilino) pyridine derivative useful as an antiviral agent, and more particularly, to the proliferation inhibitory effect of Hepatitis C virus (HCV) represented by the following Chemical Formula 1 The present invention relates to a novel 2- (4-substituted-anilino) pyridine derivative, a pharmaceutically acceptable salt thereof, a method for preparing the same, and a pharmaceutical composition for an antivirus comprising the compound as an active ingredient. The novel 2- (4-substituted-anilino) pyridine derivatives represented by the general formula (1) according to the present invention are excellent in inhibiting the proliferation of hepatitis C virus (HCV) and have low toxicity, thus preventing hepatitis C. It can be usefully used as a therapeutic agent.

  In Formula 1, R represents a heterocyclic alkylamino group or a substituted phenylpiperazinyl group.

Description

2- (4-substituted-anilino) pyridine derivatives, preparation method thereof and pharmaceutical composition for antiviral comprising same {2- (4-Substituted-anilino) pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same}

The present invention relates to a 2- (4-substituted-anilino) pyridine derivative useful as an antiviral agent, and more particularly, represented by the following Chemical Formula 1 having an excellent antiproliferative effect of Hepatitis C virus (HCV). The present invention relates to a novel 2- (4-substituted-anilino) pyridine derivative, a pharmaceutically acceptable salt thereof, a method for preparing the same, and a pharmaceutical composition for an antivirus comprising the compound as an active ingredient.

[Formula 1]

In Formula 1, R represents a heterocyclic alkylamino group or a substituted phenylpiperazinyl group.

Hepatitis C virus (HCV: hereinafter referred to as "HCV") is a major factor in non-A and non-B viral hepatitis, mainly transfusion and regionally specific infections. (community-acquired) In HCV infection, about 80% of the signs of the disease progress to chronic hepatitis and about 20% to acute hepatitis, which causes cirrhosis, and then to liver cancer. According to a recent report, more than 200 million people are infected with HCV worldwide, estimated at more than 4.5 million people in the United States (up to 15 million), and more than 5 million HCV patients in Europe alone. It is known to be.

HCV is a virus with a membrane belonging to a family of Flaviviruses different from HBV. The membrane contains about 9.5 kb of (+)-RNA (single stranded positive-sense RNA) into the genome, and the RNA genome is divided into 5 long and 3 'nonparallel segments and one long open reading frame (ORF). Consists of. The ORF is expressed as a polyprotein consisting of 3,010 to 3,040 amino acids by enzymes of the host cell, and three structural proteins and six kinds of proteins by the host cell and the virus's own protease. It is divided into nonstructural proteins. Conserved regions exist at the 5 'and 3' end of the genome, respectively, which are thought to play an important role in viral protein formation and RNA replication.

One long ORF is expressed as a polyprotein and can be expressed through the transcriptional or posttranslational processing, which is the structural antigen core antigen protein (core), the surface antigen protein (E1, E2), and the nonstructural protein NS2 (protein). Degrading enzyme, protease), NS3 (serine protease serine protease, helicase helicase), NS4A (cofactor serine protease cofactor), NS4B (protease cofactor, resistance related), NS5A, NS5B (RNA polymerase RdRp, RNA dependent It is divided into RNA polymerase and the like and each acts on virus growth. In the case of structural proteins, they are divided into parts of core, E1, and E2 by signal peptidase of the host cell, and in the case of non-structural proteins, the virus's own serine protease (NS3) and cofactor (cofactor) , NS2, NS4A, NS4B). The central antigenic protein of the structural protein, together with the surface antigenic protein, constitutes the capsid of the virus, and nonstructural proteins such as NS3 and NS5B play an important role in the replication of viral RNA (Bartenschager, R., 1997, Molecular targets in inhibition of hepatitis C virus replication, Antivir. Chem. Chemother. 8: 281-301).

At the 5 'and 3' end portions of the viral RNA, there is an untranslational region (UTR), which is conserved in the same way as other flaviviruses, and is known to play an important role in the growth of the virus. At the 5 'end, there is a 5'-UTR consisting of 341 nucleotides, which consists of four stem and loop (I, II, III, IV) structures, which are responsible for the translation process for protein expression. It acts as an internal ribosome entry site (IRES) necessary for. In particular, stem III, which has the largest and most stable structure and has a conserved sequence, has been reported as the most essential part for ribosomal attachment. In addition, the transcription process is initiated from the AUG present in the single RNA portion of stem IV and it is known that the protein of the virus is expressed (see Stanley, M. Lemon and Masao Honda, 1997, Internal ribosome entry sites within the RNA genomes of hepatitis C virus and other Flaviviruses, seminars in Virology 8: 274-288).

In addition, there is a 3'-UTR composed of 318 nucleotides at the 3 'end, which is known to play an important role in initiating the binding of NS5B, an enzyme essential for RNA replication. This part can be divided into three different parts depending on the nucleotide sequence and the three-dimensional structure. That is, it is divided into -X-tail-5 'portion consisting of 98 nucleotides (98nt.) From the 5' end, -poly (U)-portion continuously presenting UTP, and three portions of 3'-UTR-. . X-tail-5 'is composed of 98 nucleotides with very conserved nucleotide sequence, and consists of three stem and loop structure and has a stable three-dimensional structure. In addition, the -poly (U)-moiety has been reported to facilitate the action of RNA polymerase by inducing pyrimidine tracks. In addition, the 3'-UTR part has a three-dimensional structure of the loop plays an important role in the attachment of NS5B, but the structure is not stable. This 3 'end region is known to form an essential structure for the attachment of NS5B when RNA replication begins (see Yamada et al ., 1996, Genetic organization and diversity of the hepatitis C virus genome, Virology 223: 255-281). .

Among the various enzymes of HCV, NS5B is very important because it directly affects RNA replication. NS5B is an enzyme of 591 amino acids and has a molecular weight of 65 kDa. According to the amino acid sequence it can be divided into six genotypes, including 1a and 1b, has been reported to have the activity of RNA polymerization and terminal transferase (terminal transferase). In the NS5B enzyme, there are two regions of the RBD (RNA-binding domain) 1 and 2 which are essential for the attachment of the RNA required for the reaction. RBD1 exists between amino acids 83 and 194 and RBD2 exists between amino acids 196 and 298. Essential amino acids for RNA attachment and activity include asp 220, 282 gly, 317 gly, 318 asp, 319 asp and 346 lys. In addition, the enzyme can proceed polymerization without the presence of other primers in the presence of its own RNA template (Lohmann, V. et al ., 1997, Biochemical properties of hepatitis C virus NS5B RNA dependent). RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity, J. virol. 71: 8416-8428).

HCV was isolated from the RNA genome by molecular cloning in 1989 (see Choo, QL, et al. , 1989, Isolation of a cDNA clone derived from a blood-borne non-A, non-B). viral hepatitis genome.Science 244: 359-362). Since then, molecular and biological research on HCV has been conducted, but there are limitations due to the lack of an effective cell culture system and animal model. Recently, however, a hepatoma cell line capable of stably replicating HCV has been developed to solve some of these problems (see: Lohmann, V., F. Korner, JO Koch, U. Herian, L. Theilmann, R. Bartenschlager, 1999, Replication of subgenomic hepatitis c virus RNAs in a hepatoma cell line.Science 285: 110-113).

To date, no excellent vaccine or effective treatment for HCV exists. Therefore, many pharmaceutical companies and research institutes around the world are developing HCV therapy. HCV has a more even distribution worldwide than HBV, and the rate of liver cirrhosis and metastasis to liver cancer is much higher than that of HBV. And although the rate of progression to chronic hepatitis is high, studies on these infection mechanisms are still ongoing. In addition, hepatitis C can be infected not only by transfusion but also by intravenous drug use or tattooing, but mainly by direct blood contact. However, in 40% to 50% of cases, the path of infection is not related to the known factors, and when hepatitis is developed by HCV through these pathways, most patients progress to chronic and then develop cirrhosis, which has a great involvement in developing liver cancer. I think there will be. Therefore, the development of effective vaccines and treatments is urgently needed. HCV has various genotypes and mutations between strains.In case of chronic hepatitis caused by HCV, reinfection and coinfection are caused by genetic variants. You may have a back. As a result, the development of an effective vaccine for HCV was difficult. Although alpha-interferon is used as a treatment method, the effect varies depending on the genotype of HCV and in most cases recurs when discontinued. Developing one could be one of the treatments. The best targets in these studies are HCV NS3 protease / helicase and NS5B RNA polymerase. These enzymes are essential for their proliferation but are not necessary for host cells, and thus are useful for the development of anti-HCV agents. Therefore, since NS5B (RNA dependent RNA polymerase (replicase)) of HCV is an enzyme of HCV essential for proliferation, it may be a good target for treatment to inhibit the growth of HCV.

Until now, it is difficult to prevent vaccines, and treatment with α-interferon and ribavirin has been carried out, but the effect is low because the treatment rate is low and side effects appear. In the case of interferon therapy, there is about 25% of cases that do not respond at all, and about 25% of cases temporarily relapse. In about 50% of patients, ALT levels remain normal and HCV RNA becomes negative until the end of treatment. Of these, about 50% relapse within 3-6 months after the end of treatment. It shows a sustained response where the therapeutic effect is maintained. In addition, there are six genotypes (subtypes) in the hepatitis C virus, the most type 1b in Korea does not respond better to the treatment of interferon than type 2,3. For this reason, the combination therapy with ribavirin has been shown to be twice as effective, but the use of ribavirin alone alone has little effect, and red blood cells are destroyed, resulting in side effects such as anemia. It is known to prescribe it when there is no response or relapse in treatment. Therefore, until now, antiviral drugs for treating hepatitis C by specifically acting on HCV and inhibiting proliferation have not been developed.

Accordingly, the present invention searches for a substance that inhibits the activity of recombinant expressed HCV RNA polymerase (NS5B, RNA polymerase), thereby exhibiting a good antiviral activity against HCV, nontoxic compounds with low toxicity and side effects (nonnuleoside) The purpose was to develop.

Accordingly, the present inventors have tried to develop a compound exhibiting excellent antiviral activity against HCV for the purpose of developing a novel hepatitis C therapeutic agent with low side effects and toxicity, and low resistance virus appearance. The present invention was completed by synthesizing a-(4-substituted-anilino) pyridine derivative and showing that the substance was excellent in inhibiting the growth of HCV.

It is an object of the present invention to provide novel 2- (4-substituted-anilino) pyridine derivatives, pharmaceutically acceptable salts thereof and methods for their preparation. Another object of the present invention is to provide a pharmaceutical composition for the treatment and prevention of hepatitis C, the compound as an active ingredient and less side effects and economical.

In order to achieve the above object, the present invention provides a novel 2- (4-substituted-anilino) pyridine derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1, R represents a heterocyclic alkylamino group or a substituted phenylpiperazinyl group.

Preferably, in Chemical Formula 1, R is a (2-pyridyl) methylamino group, 2- (morpholin-4-yl) ethylamino group, 4- (4-hydroxyphenyl) piperazin-1-yl group or 4- It is a (2-hydroxyphenyl) piperazin-1-yl group.

The compound of formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Compounds of formula (1) may form pharmaceutically acceptable acid addition salts according to methods conventional in the art. Organic acids and inorganic acids may be used as the free acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, Fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid or aspartic acid can be used. .

In addition, the present invention provides a method for preparing a 2- (4-substituted-anilino) pyridine derivative represented by the following Scheme 1.

In Scheme 1, R represents a heterocyclic alkylamino group or a substituted phenylpiperazinyl group.

In the preparation method of the present invention, as shown in Scheme 1, (1) by reacting 2,6-dichloro-3-nitropyridine of the formula (2) with 4- (4-morpholino) aniline of the formula (3) Preparing 3-nitro-2- [4- (4-morpholino) anilino] -6-chloropyridine of 4 (step 1); And (2) reacting the synthetic intermediate of Formula 4 prepared in Step 1 with a heterocyclic alkylamine or substituted phenylpiperazine to prepare 2- (4-substituted-anilino) pyridine derivative of Formula 1 It consists of a step (step 2).

2,6-dichloro-3-nitropyridine, 4- (4-morpholino) aniline, heterocyclicalkylamine, substituted phenylpiperazine, etc. used as starting materials and reactants in Scheme 1 are commercially available. It can be easily purchased and used as a material to be used, the reagent such as heterocyclic alkylamine or substituted phenyl piperazine used in the step 2 is a material for introducing a substituent to the target compound, appropriate reagents depending on the type of substituent It can be selected, which can be easily selected and used by those of ordinary skill in the art.

In more detail the preparation method of step 1 and step 2, triethyl in an organic solvent such as methanol, ethanol, isopropanol, dichloromethane, chloroform, acetonitrile, N, N- dimethylformamide, acetone, etc. General organics such as amines, N, N -diisopropylethylamine, N -methylmorpholine, 1-methylpiperidine, pyridine, 2,6-lutidine, 4-dimethylaminopyridine, N, N -dimethylaniline and the like In the presence of a base, it progresses well within 2 to 20 hours in the temperature range of 20 ° C-65 ° C.

In another aspect, the present invention provides a pharmaceutical composition for treating or preventing hepatitis C, comprising a 2- (4-substituted-anilino) pyridine derivative of Formula 1 and / or a pharmaceutically acceptable salt thereof as an active ingredient. do.

The compound of formula 1 of the present invention can be administered orally or parenterally during clinical administration as a therapeutic agent for hepatitis C, and can be used in the form of general pharmaceutical preparations. That is, the compound of formula 1 of the present invention may be administered in various dosage forms, orally and parenterally (preferably as an injection) during actual clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents. And diluents or excipients such as disintegrants and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, or the like in one or more compounds of formula (I). Sucrose (Sucrose) or lactose (Lactose), gelatin and the like are mixed and prepared. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions and the like. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used.

The effective dose of the compound of formula 1 of the present invention may be appropriately selected in consideration of sex, age, patient's condition, etc., but is generally 10-1000 mg / day, preferably 20-500 mg / day in adults, It may be administered in divided doses 1-3 times a day.

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

Preparation Example 1 Preparation of 3-nitro-2- [4- (4-morpholino) anilino] -6-chloropyridine.

To 60 ml of methanol, 3.86 g of 2,6-dichloro-3-nitropyridine, 3.57 g of 4- (4-morpholino) aniline, and 3.1 ml of triethylamine were added sequentially, and reacted at 25 ° C-30 ° C for 15 hours. I was. After cooling to 20 ° C., stirring for 2 hours, filtration and washing with 20 ml of methanol, the obtained crystals were vacuum dried at 30 ° C.-40 ° C. to obtain 6.3 g (yield 94%) of the title compound.

  m.p. : 164-165 ℃

¹ H-NMR (CDCl ₃ ), ppm: 3.17 (t, 4H), 3.88 (t, 4H), 6.72 (d, 1H), 6.93 (d, 2H), 7.52 (d, 2H), 8.42 (d, 1H)

Example 1 Preparation of 3-nitro-2- [4- (4-morpholino) anilino] -6-[(2-pyridyl) methylamino] pyridine.

To 40 ml of methanol, 2 g of 3-nitro-2- [4- (4-morpholino) anilino] -6-chloropyridine and 1.95 g of 2- (aminomethyl) pyridine prepared in Preparation Example 1, and tri 0.9 ml of ethylamine was added sequentially, followed by heating for 12 hours at 55 ° C-60 ° C. After adding 5 ml of water, the mixture was slowly cooled, stirred at 20 ° C. for 2 hours, filtered, and washed with a mixed solvent of 10 ml of methanol and 3 ml of water. The solid was vacuum dried at 35 ° C.-45 ° C. to obtain 2.1 g (yield 86%) of the title compound.

  m.p. : 163-164 ℃

¹ H-NMR (CDCl ₃ ), ppm: 3.16 (t, 4H), 3.88 (t, 4H), 4.69 (d, 2H), 5.99 (d, 1H), 6.63 (s, 1H), 6.88 (m, 2H), 7.22 (m 2H), 7.51 (d, 2H), 7.62 (t, 1H), 8.20 (d, 1H), 8.55 (d, 1H), 10.78 (s, 1H)

Example 2 Preparation of 3-nitro-2- [4- (4-morpholino) anilino] -6- [2- (morpholin-4-yl) ethylamino] pyridine

To 40 ml of methanol, 1.8 g of 3-nitro-2- [4- (4-morpholino) anilino] -6-chloropyridine and 2.1 g of 4- (2-aminoethyl) morpholine prepared in Preparation Example 1 And 0.9 ml of triethylamine was added sequentially, and it heated and reacted at 50 degreeC-55 degreeC for 15 hours. It was cooled slowly and stirred at 20 ° C. for 3 hours, then filtered and washed with 8 ml of methanol. The solid was vacuum dried at 30 ° C.-40 ° C. to obtain 2.03 g (yield 88%) of the title compound.

  m.p. : 199-200 ℃

¹ H-NMR (CDCl ₃ ), ppm: 2.48 (brs, 4H), 2.58 (t, 2H), 3.15 (m, 4H), 3.51 (brs, 2H), 3.71 (t, 4H), 3.86 (t, 4H), 5.83 (s, 1H), 5.90 (m, 1H), 7.50 (m, 2H), 8.20 (d, 1H), 10.83 (s, 1H)

Example 3 Preparation of 3-nitro-2- [4- (4-morpholino) anilino] -6- [4- (4-hydroxyphenyl) piperazin-1-yl] pyridine

To 50 ml of methanol, 2 g of 3-nitro-2- [4- (4-morpholino) anilino] -6-chloropyridine and 1.13 g of 4- (1-piperazino) phenol prepared in Preparation Example 1 And 1 ml of triethylamine was added sequentially, and it heated and reacted at 55 degreeC-60 degreeC for 4 hours. It was cooled slowly and stirred at 25 ° C. for 2 hours, then filtered and washed with 10 ml of methanol. The solid was vacuum dried at 30 ° C.-40 ° C. to obtain 2.71 g (yield 95%) of the title compound.

  m.p. : 225-227 ℃

¹ H-NMR (CDCl ₃ + DMSO-d ₆ ), ppm: 3.09 (brs, 4H), 3.16 (t, 4H), 3.86 (t, 8H), 6.19 (d, 1H), 6.77 (m, 4H) , 6.92 (d, 2H), 7.48 (d, 2H), 8.25 (m, 1H), 8.44 (s, 1H), 10.62 (s, 1H)

Example 4 Preparation of 3-nitro-2- [4- (4-morpholino) anilino] -6- [4- (2-hydroxyphenyl) piperazin-1-yl] pyridine

The target compound was obtained in the same manner as in Example 3 using 2- (1-piperazino) phenol instead of 4- (1-piperazino) phenol.

  Yield: 96%

  m.p. : 186-189 ℃

¹ H-NMR (CDCl ₃ ), ppm: 2.93 (t, 4H), 3.14 (t, 4H), 3.87 (d, 8H), 6.16 (d, 1H), 6.85 (m, 4H), 6.97 (m, 2H), 7.46 (d, 2H), 8.30 (d, 1H), 10.59 (s, 1H)

Experimental Example 1 In Vitro for HCV RNA Polymerase (RNA-dependent RNA Polymerase, NS5B)

( in vitro ) Inhibitory activity investigation

In vitro experiments were performed to investigate the inhibitory effect of the compounds of the present invention on HCV RNA dependent RNA polymerase.

Preparation of Recombinant HCV RNA Polymerase

HCV RNA polymerase was prepared by the following method. First, cDNA was obtained from the blood of HCV-1b hepatitis C virus patient, and then a recombinant transfer vector inserted into the baculovirus transition vector pVLHIS was prepared by amplifying the NS5B (1773bps) region by PCR. The prepared transfection vector was cotransfected with an insect cell (Sf 9 cell line) together with a wild-type AcNPV vector to produce a recombinant baculovirus with histidine tagged recombinant vector pVLHIS-NS5B. The recombinant baculovirus thus prepared was infected with sufficiently cultured insect cells, incubated for 3-4 days in Grace 'medium to which 10% FBS was added, and only infected cells were centrifuged. The obtained cells were washed three times with PBS, sonicated, and subjected to an affinity column chromatography using NI-NTA His bind resin (Novagen) to prepare pure NS5B protein.

    RNA template preparation comprising HCV 3 'terminus (3'-UTR)

RNA template containing HCV 3'-UTR was prepared by the following method. First, cDNA was obtained from the HCV RNA obtained from the blood of hepatitis C patients by PCR, and then DNA fragments containing the 3'-UTR region were obtained using restriction enzyme Eco Ri. RNA from the 3'-UTR was prepared from the DNA through in vitro transcription.

    In Vitro Inhibition of Compounds of the Invention on Recombinant HCV RNA Polymerase

The method for measuring the in vitro inhibitory activity of the compounds of the present invention on the HCV RNA polymerase used in the present invention is as follows. Prepare wells coated with streptavidin for the sample to be measured, and firstly, in each well, 2X reaction buffer [50 mM Tris-Cl (pH 7.5), 100 mM NaCl, 10 mM MgCl ₂ , 20 mM KCl , 1mM EDTA, 1mM DTT] were added thereto, and 200ng of the purified HCV RNA polymerase was added 10µl each. Then, the final test substance was measured to have a final concentration of 10, 1, 0.1, 0.01µg / ml, respectively. 5 μl each was added, and finally a reaction mixture of DIG- (digoxigenin) -UTP, biotin-UTP, ATP, CTP, GTP, and UTP was mixed as a nucleotide for reaction with HCV 3'-UTR RNA, an RNA template. 10 μl each was added, followed by reaction at 22 ° C. for 60 minutes. The RNA is produced by the activity of HCV polymerase, which contains UTP with biotin attached, so the new RNA binds to the well-coated streptavidin. After the reaction, 200 μl of washing buffer (washing buffer, pH 7.0, Roche) was added to each well to remove impurities remaining unreacted, and then washed three times. Then, 100 μl of a secondary antibody, anti-DIG-POD (peroxidase, Roche) was added thereto, reacted at 37 ° C. for 1 hour, and each well was washed again with a washing buffer. Finally, 100 μl of ABTS ^R (Roche), which is a substrate of POD, was added and reacted for 15-30 minutes, and then the absorbance (OD value) was measured at 405 nm using an ELISA reader (Bio-Tek instrument). At this time, the inhibitory effect on the activity of HCV polymerase was calculated according to the difference in absorbance relative to the control without the sample (positive control). The results are shown in Table 1 below.

 Test compound       Substituent (R) HCV RNA Activity inhibition rate of polymerase (%)  10 μg / ml  1 μg / ml  0.1 μg / ml 0.01 µg / ml   Example 1 (2-pyridyl) methylamino    93    65    34    23   Example 2 2- (morpholin-4-yl) ethylamino    81    54    40    15   Example 3 4- (4-hydroxyphenyl) piperazin-1-yl    95    64    50    29   Example 4 4- (2-hydroxyphenyl) piperazin-1-yl    80    52    36    12

As described above, the compounds of the present invention have an excellent effect of inhibiting the activity of HCV RNA polymerase, which plays an important role in the replication of HCV. Therefore, the compounds of the present invention can inhibit the proliferation of HCV, and thus are useful as a prophylactic and therapeutic agent for hepatitis C. Can be used.

Experimental Example 2 Cytotoxicity Experiment

In order to determine whether the compounds of Formula 1 exhibit cytotoxicity (Cytotoxicity), in vitro experiments were carried out by MTT assay generally known using Hep G2 cells. As a result, the compounds used in the experiments all had a CC50 value of 100 µg / ml or more, and were found to be very low toxicity to cells.

As described above, the novel 2- (4-substituted-anilino) pyridine derivative represented by Chemical Formula 1 according to the present invention is excellent in effect of inhibiting the proliferation of hepatitis C virus (HCV) and toxicity. As a result, it can be usefully used as a prophylactic and therapeutic agent for hepatitis C.

Claims (4)

2- (4-substituted-anilino) pyridine derivatives represented by the following formula (1) and pharmaceutically acceptable salts thereof. [Formula 1] Wherein R is (2-pyridyl) methylamino, 2- (morphyrin-4-yl) ethylamino or hydroxyphenylpiperazinyl. The compound of claim 1, wherein R is (2-pyridyl) methylamino, 2- (morpholin-4-yl) ethylamino, 4- (4-hydroxyphenyl) piperazin-1-yl or 4- (2 2- (4-substituted-anilino) pyridine derivatives characterized by hydroxyphenyl) piperazin-1-yl and pharmaceutically acceptable salts thereof. In reaction scheme 1, 2,6-dichloro-3-nitropyridine of formula (2) is reacted with 4- (4-morpholino) aniline of formula (3) to yield 3-nitro-2- [4- (4-mo of formula (4). Preparing lipolino) anilino] -6-chloropyridine; And reacting the synthetic intermediate of formula (4) prepared in the step with 2- (aminomethyl) pyridine, 4- (2-aminoethyl) morphyrin or hydroxyphenylpiperazine to form 2- (4-substituted- A process for preparing a 2- (4-substituted-anilino) pyridine derivative, characterized by preparing an anilino) pyridine derivative. Scheme 1 Wherein R is (2-pyridyl) methylamino, 2- (morphyrin-4-yl) ethylamino or hydroxyphenylpiperazinyl. A pharmaceutical composition for preventing the treatment of hepatitis C, comprising the 2- (4-substituted-anilino) pyridine derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.