KR100566189B1 - Novel 5-pyrimidinecarboxamide derivatives and pharmaceutical compositions thereof - Google Patents

Abstract

The present invention relates to a novel 5-pyrimidinecarboxamide derivative, a preparation method thereof, and a pharmaceutical composition comprising the same. Specifically, the 5-pyrimidinecarboxamide derivative represented by the following Chemical Formula 1, which is a non-nucleic acid compound, is HBV. (Hepatitis B Virus) As well as the proliferation of the HIV (Human Immunodeficiency Virus) proliferation, it can be useful as a therapeutic and prophylactic agent for hepatitis B and AIDS.

Wherein R ₁ , R ₂ , R ₃ and n are as described in the specification.

Description

Novel 5-pyrimidine carboxamide derivatives and pharmaceutical compositions             

The present invention relates to a novel 5-pyrimidinecarboxamide derivative, and more particularly, to a novel 5-pyrimidinecarboxamide derivative represented by the following Chemical Formula 1, which is excellent in inhibiting the proliferation of HBV and HIV, and a pharmaceutical thereof The present invention relates to a salt, a method for preparing the same, and an antiviral composition comprising the compound as an active ingredient.

Formula 1

In Chemical Formula 1,

R ₁ is hydroxy; C ₁ -C ₅ linear or pulverized alkyl group; C ₁ -C ₅ linear or pulverized alkoxy group; C ₂ -C ₆ linear or pulverized hydroxyalkyl group; C ₂ -C ₆ dialkylamino group; A C _{2 to} C ₆ straight or pulverized alkyl group substituted with a C _{2 to} C ₅ alkoxycarbonyl group and / or a hydroxy group; Or is optionally substituted with C ₁ ~C ₃ alkyl group, and a N, O, S, a saturated or unsaturated 5-atom or 6-atom heterocyclic compound containing 1 to 3 heteroatoms selected from substituted, R ₁ is asymmetrical With or without carbon,

R ₂ is H; Or a C ₁ -C ₄ straight or crushed alkyl group,

Or R ₁ and R ₂ are bonded to each other to form a saturated 5- or 6-membered heterocyclic compound, wherein the heterocycle includes 1 to 3 heteroatoms selected from N, O, and S and the heterocycle is substituted. Unsubstituted or substituted with a C ₁ -C ₅ straight or pulverized alkyl group or a C ₂ -C ₅ straight or pulverized hydroxyalkyl group,

R ₃ is indazol-5-yl; Or indazol-6-day,

n is an integer of 0-4.

Hepatitis B virus (HBV; hereinafter referred to as "HBV") is a pathogen that causes chronic or acute hepatitis and worsens liver cirrhosis and liver cancer. It is estimated (Tiollais & Buendia, Sci. Am. , 264, 48, 1991). Therefore, for the purpose of treatment and prevention of hepatitis B, many studies have been conducted on the relationship between HBV and liver disease, including molecular biological characteristics of HBV, and various vaccines and diagnostic reagents for hepatitis B have been developed. In addition, efforts are underway to develop treatments for hepatitis B.

The genome of HBV is polymerase gene (P; polymerase), surface antigen gene (S; surface protein; pre-S1, pre-S2, S), central antigen gene (C; core protein; pre-C, C), X It consists of four genes, including the protein gene. Among them, polymerase, surface antigen, and central antigen genes express structural proteins, and X protein genes are known to express regulatory proteins.

The HBV polymerase gene accounts for 80% of the entire viral genome and produces a 94kD protein consisting of 845 amino acids, which contains a set of functions necessary for the replication of the viral genome. Ie) protein primer, ii) RNA dependent DNA polymerase (RT), iii) DNA dependent DNA polymerase (DDDP), iii) RNA degradation Enzyme (RNase H) function and the like are present in one polypeptide. Kaplan et al. Described the reverse transcription activity of polymerase protein for the first time, and many studies on the replication mechanism of HBV have been made.

In HBV, surface antigen proteins outside the virion are recognized by hepatocyte-specific receptors and enter into the hepatocytes. At this time, the rest of the incomplete double-stranded DNA is synthesized by HBV polymerase activity, and the HBV DNA genome is synthesized. Is completed. The completed HBV DNA genome produces mRNAs such as pre-genomic mRNAs, central antigens (C), surface antigens (S), and regulatory proteins (X) by intracellular RNA polymerase activity. Viral proteins are made from these mRNAs, and in particular, polymerase proteins play a role in synthesizing the viral genome and form structures called replicatics with central antigen proteins and genome mRNAs. This is called encapsidation, and the polymerase protein is easily encapsidated by nucleic acid affinity at a site where glutamic acid is repeated at the 3′-end. When a replicasome is formed, (-) DNA chain is synthesized by reverse transcription activity of HBV polymerase protein, and (+) DNA chain is synthesized by DNA dependent DNA polymerase activity, which in turn produces genome mRNAs. This repetition maintains a pool of 200-300 or more genomic DNA in the cell (Tiollais and Buendia, Scientific American , 264: 48-54, 1991).

HBV and HIV are different kinds of viruses, but they have a common replication process. That is, the reverse transcription process in which transcription from viral RNA to DNA is performed and the process of decomposing and erasing the RNA portion of the RNA-DNA hybrid generated by reverse transcription is common.

HBV inhibitors of nucleic acid-based compounds, such as lamivudine and famvir, which have recently been developed as treatments for Acquired Immune Deficiency Syndrome (AIDS) or shingles infection As usefulness (Gerin J. L, Hepatology , 14: 198-199, 1991; Lok A. .SP, J. Viral Hepatitis , 1: 105-124, 1994; Dienstag, JL et al ., New England Journal of Medicine , 333: 1657-1661, 1995). However, these nucleic acid-based compounds are very expensive and put a high economic burden on patients, and furthermore, since nucleic acid-based compounds inherently have serious problems in side effects such as toxicity, emergence of resistant virus and relapse after discontinuation of drug administration. It is known that it is unsuitable as a hepatitis agent. Therefore, efforts have been made to develop a hepatitis B therapeutic agent among non-nucleosides compounds, and quinolone compounds having antiviral activity against HBV (European Patent Nos. 563732 and 563734), iridoid compounds (Korean Patent Publication No. 94-1886) and terephthalic acid amide derivatives (Korean Patent Application Nos. 96-72384, 97-36589, 99-5100) and the like have been reported. However, despite many efforts, no clear treatment for hepatitis B has been developed so far, and the treatment of hepatitis B is mainly dependent on symptomatic therapy.

On the other hand, AIDS is a disease that spreads throughout the body due to the distinctly low cellular immune function of the body, which is not seen in ordinary people, and is the main attack of HIV (Human Immunodeficiency Virus, or "HIV"). The goal is known to be helper T cells, one of the T cells that regulate immune function. When secondary T cells become infected by HIV and cause necrosis, the body's immune function is not functioning properly, resulting in an immunodeficiency condition, which causes fatal infections and malignant tumors. Since it was first discovered in the United States in 1981, more than 850,000 people have been reported in 187 countries in 1993 (the World Health Organization's late 1993 report). Furthermore, the World Health Organization reports that by 2000, 30 million to 40 million more will be infected and 10 million to 20 million of them will develop.

Drugs that inhibit the proliferative process of HIV are currently the most widely used in the treatment of AIDS, including Zidovudine (ZDV), formerly named Azidothymidine (AZT), which was first developed in 1987, and have side effects or treatment effects Alternative medicines include Didanosine (ddI), developed in 1991, and Zalcitabine (ddC), licensed for use in combination with zidovudine in 1992. These drugs have the effect of alleviating the symptoms of AIDS patients, slowing the transition of HIV infected people to AIDS, or slightly extending their survival. However, there is no cure and resistance and side effects are a problem.

Accordingly, the present inventors have endeavored to develop a non-nucleic acid compound exhibiting excellent antiviral activity against HBV for the purpose of developing a new hepatitis B drug having low side effects and toxicity, and low resistance virus appearance. A novel 5-pyrimidinecarboxamide derivative was synthesized and the present invention was completed by revealing that the substance was excellent in inhibiting HBV growth as well as in inhibiting HIV growth.

An object of the present invention is to provide a novel 5-pyrimidinecarboxamide derivative, a pharmaceutically acceptable salt thereof, and a method for preparing the same, which are excellent in inhibiting the growth of HBV and HIV.

It is also an object of the present invention to provide a pharmaceutical composition for the treatment and prevention of hepatitis B or AIDS, comprising the compound as an active ingredient and having low side effects and economical.

In order to achieve the above object, the present invention provides a novel 5-pyrimidinecarboxamide derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof.

Formula 1

In Chemical Formula 1,

R ₁ is hydroxy; C ₁ -C ₅ linear or pulverized alkyl group; C ₁ -C ₅ linear or pulverized alkoxy group; C ₂ -C ₆ linear or pulverized hydroxyalkyl group; C ₂ -C ₆ dialkylamino group; A C _{2 to} C ₆ straight or pulverized alkyl group substituted with a C _{2 to} C ₅ alkoxycarbonyl group and / or a hydroxy group; Or is optionally substituted with C ₁ ~C ₃ alkyl group, and a N, O, S, a saturated or unsaturated 5-atom or 6-atom heterocyclic compound containing 1 to 3 heteroatoms selected from substituted, R ₁ is asymmetrical With or without carbon,

R ₂ is H; Or a C ₁ -C ₄ straight or crushed alkyl group,

Or R ₁ and R ₂ are bonded to each other to form a saturated 5- or 6-membered heterocyclic compound, wherein the heterocycle includes 1 to 3 heteroatoms selected from N, O, and S and the heterocycle is substituted. Unsubstituted or substituted with a C ₁ -C ₅ straight or pulverized alkyl group or a C ₂ -C ₅ straight or pulverized hydroxyalkyl group,

R ₃ is indazol-5-yl; Or indazol-6-day,

n is an integer of 0-4.

In Formula 1, when R ₁ and R ₂ are bonded to each other to form a 5 or 6 atom heterocyclic compound, wherein the heterocycle includes 1 to 3 heteroatoms selected from N, O, and S, n is 0. This heterocycle can also be unsubstituted or substituted with C ₁ -C ₅ straight or pulverized alkyl or C ₂ -C ₅ straight or pulverized hydroxyalkyl groups.

In addition, an optical isomer of a compound of formula (I) is R or S if R ₁ contains an asymmetric carbon in the formula 1, the invention includes all of these optical isomers and racemic mixtures.

Indazol-5-yl and indazol-6-yl in the present invention are represented by the following formula (2) and (3).

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, and fumaric acid may be used as the organic acid. (fumaric acid), formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galluxuronic acid, embonic acid, glutamic acid or aspartic acid Can be used.

In addition, the present invention provides a method for preparing a 5-pyrimidinecarboxamide derivative represented by the following Scheme 1.

In Scheme 1, R ₁ , R _2, R ₃ and n are as defined in formula (1).

The manufacturing method of the present invention

1) 4-chloro-2-methylthio-5-pyrimidinecarboxylic acid ethyl ester ( 2 ) and 5-aminoindazole or 6-aminoindazole ( 3 ) are reacted in a suitable temperature and a suitable solvent in the presence of a base Preparing 5-pyrimidinecarboxylic acid ethyl ester derivative ( 4 ) (step 1);

2) preparing a 5-pyrimidinecarboxylic acid derivative ( 5 ) by hydrolyzing the 5-pyrimidinecarboxylic acid ethyl ester derivative ( 4 ) prepared in step 1 under alkaline conditions (step 2); And

3) the 5-pyrimidinecarboxylic acid derivative ( 5 ) prepared in step 2 was activated with a Vilsmeier intermediate in the presence of N, N -dimethylformamide (DMF) and thionyl chloride (SOCl ₂ ). And a 5-pyrimidinecarboxamide derivative of Formula 1 (corresponding to Formula 7 in Scheme 1) by reacting with an amine compound ( 6 ) (Step 3).

4-chloro-2-methylthio-5-pyrimidinecarboxylic acid ethyl ester ( 2 ), 5-aminoindazole or 6-amino, used as starting materials and reactants in steps 1, 2 and 3 above The sol ( 3 ) and the amine compound ( 6 ) can be easily purchased and used as commercially available materials.

In addition, the amine compound ( 6 ) of step 3 is a substance for introducing a substituent (-NR _2- (CH ₂ ) _n -R ₁ ) to the compound of formula ( ₁ ), depending on the type of the substituent, a suitable amine compound ( 6 ) It can be selected and can be easily selected and used by those of ordinary skill in the art.

In more detail the step 1, an organic base may be used as a base, for example triethylamine, N, N -diisopropylethylamine, N -methylmorpholine, N -methylpiperidine Preference is given to using common tertiary organic bases such as, 4-dimethylaminopyridine, N, N -dimethylaniline, 2,6-lutidine, pyridine and the like.

In addition, when the step 2 is described in more detail, the alkaline substance used for hydrolysis is preferably sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate and the like. At this time, 5-pyrimidinecarboxylic acid derivative ( 5 ) can be produced almost quantitatively through hydrolysis.

It is preferable to use a mixed solvent of water and alcohols such as methanol and ethanol as the reaction solvent.

It is preferable that reaction temperature is 30-60 degreeC, and it is preferable to make reaction for 30 minutes-3 hours.

In addition, the step 3 will be described in more detail. The N, N -dimethylformamide and thionyl chloride are heated at a temperature in the range of 30 to 50 ° C. to generate a Vilsmeier Reagent, whereby 5-pyri After activating the midine carboxylic acid derivative ( 5 ), it is reacted with the amine compound ( 6 ) at a temperature of 0 to 20 ° C to prepare a 5-pyrimidinecarboxamide derivative of the formula (1).

At this time, as a reaction solvent, an aprotic solvent which cannot dissociate protons is preferable, and for example, chloroform, methylene chloride, acetonitrile, tetrahydrofuran, ether and the like are preferable.

The present invention also provides a pharmaceutical composition for treating or preventing hepatitis B, comprising 5-pyrimidinecarboxamide derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect, the present invention provides a pharmaceutical composition for the treatment or prevention of acquired immune deficiency syndrome comprising the 5-pyrimidinecarboxamide derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

5-pyrimidinecarboxamide derivatives represented by the general formula (1) of the present invention is a replication process which is common in the proliferation process of HBV and HIV, ie reverse transcription from viral RNA to DNA, and the RNA portion of the resulting RAN-DNA hybrid It has a mechanism of action that inhibits the process of degradation and has proliferation inhibitory activity against HIV as well as HBV.

The compound of formula 1 may be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically, during clinical administration and may be used in the form of general pharmaceutical formulations.

Clinical use of the pharmaceutical composition of the present invention may be combined with a conventional carrier in the pharmaceutical field for oral administration of a conventional agent in the pharmaceutical field, for example, tablets, capsules, troches, solutions, suspensions, and the like. Formulations; It may be formulated into a variety of preparations, such as injectable solutions or suspensions, or injectable preparations in the form of ready-to-use injectable dry powders that can be prepared and used as injectable distilled water at the time of injection.

The effective dose of the compound of formula 1 is generally 10-500 mg / kg in adults, preferably 50-300 mg / kg, several times a day at a predetermined time interval, preferably one day, at the discretion of the doctor or pharmacist. It may be administered in 1 to 6 divided doses.

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.

Production Example 1 4- (1 H Preparation of -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid ethyl ester

To 70 ml of methanol, 5 g of 4-chloro-2-methylthio-5-pyrimidinecarboxylic acid ethyl ester, 3.15 g of 5-aminoindazole and 3.5 ml of triethylamine were added sequentially, followed by reaction at 30 ° C. for 3 hours. I was. The reaction mixture was cooled, stirred at 20 ° C. for 1 hour, filtered and washed with 20 ml of methanol. The obtained crystals were vacuum dried at 40 to 50 ° C. to obtain 6.15 g (yield 87%) of the title compound.

m.p. 199 to 201 ° C

¹ H-NMR (DMSO-d ₆ ), ppm: δ1.34 (t, 3H), 2.42 (s, 3H), 4.34 (m, 2H), 7.48 (d, 1H), 7.53 (d, 1H), 8.06 (d, 2H), 8.70 (s, 1H), 10.18 (s, 1H), 13.10 (br s, 1H)

Production Example 2 4- (1 H Preparation of -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid ethyl ester

To 70 ml of methanol was added 5 g of 4-chloro-2-methylthio-5-pyrimidinecarboxylic acid ethyl ester, 3.15 g of 6-aminoindazole and 4.2 ml of N, N -diisopropylethylamine, and then 30 The reaction was carried out at ˜35 ° C. for 4 hours. The reaction mixture was cooled, stirred at 20 ° C. for 1 hour, filtered and washed with 20 ml of methanol. The obtained crystals were vacuum dried at 40 to 50 ° C. to obtain 5.8 g (yield 82%) of the title compound.

m.p. : 212 to 214 ° C

¹ H-NMR (DMSO-d ₆ ), ppm: δ1.33 (t, 3H), 2.53 (s, 3H), 4.33 (m, 2H), 7.10 (d, 1H), 7.70 (d, 1H), 8.00 (s, 1H), 8.22 (s, 1H), 8.72 (s, 1H), 10.40 (s, 1H), 13.09 (br s, 1H)

Production Example 3 4- (1 H Preparation of -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid

To 80 ml of methanol, 5 g of 4- ( 1H -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid ethyl ester prepared in Preparation Example 1 was added thereto, and water 30 at 20 to 25 ° C. Ml and 3n 15 ml of aqueous sodium hydroxide solution was slowly added and heated to hydrolyze at 40 to 50 ° C. for 1 hour. Cool the reaction, 3N at 20 ° C An aqueous hydrochloric acid solution was slowly added to adjust pH = 5 and 100 ml of water was added slowly. The reaction was stirred at 20 ° C. for 1 h, then filtered and washed with 30 mL of water. The obtained crystals were dried in vacuo at 50 ° C. to obtain 4.44 g (yield 97%) of the title compound.

m.p. :> 270 ℃

¹ H-NMR (DMSO-d ₆ ), ppm: δ 2.45 (s, 3H), 7.49 (d, 1H), 7.53 (d, 1H), 8.05 (s, 1H), 8.10 (s, 1H), 8.68 (s, 1H), 10.50 (s, 1H), 13.09 (br s, 1H)

Production Example 4 4- (1 H Preparation of -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid

Preparation same as Preparation Example 3, except that 4- ( 1H -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxylic acid ethyl ester prepared in Preparation Example 2 was used as a starting material. The target compound (yield 95%) was obtained by the method.

m.p. :> 270 ℃

¹ H-NMR (DMSO-d ₆ ), ppm: δ2.56 (s, 3H), 7.10 (d, 1H), 7.72 (d, 1H), 8.01 (s, 1H), 8.25 (s, 1H), 8.73 (s, 1 H), 10.81 (br s, 1 H), 13.06 (br s, 1 H)

<Example 1> N Hydroxyethyl N- Methyl-4- (1 H Preparation of -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide

1.1 ml of N, N -dimethylformamide and 1.2 ml of thionyl chloride were added to 120 ml of methylene chloride, and heated to reflux for 2 hours. Then, 4- ( 1H -5-indazolylamino)-prepared in Preparation Example 3 3 g of 2-methylthio-5-pyrimidinecarboxylic acid was added and refluxed for 10 hours. The reaction was cooled and 4 ml of 2- (methylamino) ethanol was slowly added at 0-5 ° C. and stirred for 1 hour. Methanol 80ml was added to the reaction, stirred for 5 minutes, and filtered to remove impurities. The filtrate was concentrated under reduced pressure, and the solid was added to a 1: 1 mixed solvent of methanol and water, and 3N. 3 ml of aqueous sodium hydroxide solution was added, stirred for 1 hour, filtered, and washed with water. The obtained solid was recrystallized from 1: 4 mixed solvent of methylene chloride and isopropyl ether to obtain 1.61 g (yield 45%) of the title compound.

m.p. : 106 ~ 114 ℃

¹ H-NMR (DMSO-d ₆ ), ppm: δ 2.40 (s, 3H), 2.99 (s, 3H), 3.43 (br s, 2H), 3.56 (br s, 2H), 7.44 (d, 1H ), 7.50 (d, 1H), 7.93 (br s, 1H), 8.03 (s, 1H), 8.14 (s, 1H), 8.93 (br s, 1H), 13.03 (br s 1H)

Through the same synthesis method as in Example 1, the compound of Examples 2 to 27 was prepared. Table 1 shows the names, yields, solvents used for recrystallization, melting points of crystals and starting materials used for the preparation of 5-pyrimidinecarboxylic acid derivatives ( 5 ) in Examples 2 to 27 , and An amine compound ( 6 ) is shown. In addition, Table ¹ shows the ¹ H-NMR results for the compounds prepared in Examples 2 to 27.

Example number Name of the compound 5-pyrimidinecarboxylic acid derivatives (2006.01) Amine Compounds (6) Recrystallized solvent Yield (%) Melting point (℃)  2 N - hydroxyethyl - N- methyl -4- (1 H -6- indazolyl) -2-methylthio-5-pyrimidine-carboxamide Preparation Example 4 2- (methylamino) ethanol Chloroform / isopropyl ether (1: 4) 52 93-97  3 N -[( 1R ) -1- (hydroxymethyl) propyl] -4- ( 1H -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide  Preparation Example 3 ( R ) -2-amino-1-butanol Methanol / Chloroform / Isopropyl Ether (1: 2: 10)  44  219-222  4 N -[( 1R ) -1- (hydroxymethyl) propyl] -4- ( 1H -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 4 ( R ) -2-amino-1-butanol Methanol / Isopropyl Ether (1: 5) 65 234-237  5 4- (1 H-5-indazolyl-amino) - N - (2- methoxyethyl) -2-methylthio-5-pyrimidine-carboxamide Preparation Example 3 2-methoxyethylamine Acetone / isopropyl ether (1: 4) 72 214-217  6 4- (1 H -6- indazolyl amino) - N - (2- methoxyethyl) -2-methylthio-5-pyrimidine-carboxamide Preparation Example 4 2-methoxyethylamine Acetone / isopropyl ether (1: 4) 75 225-228  7 N , N -dimethyl-4- ( 1H -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 3 Dimethylamine ethanol solution (33%) Methanol / ethyl acetate / hexanes (1: 5: 5) 41 101-105

Example number Name of the compound 5-pyrimidinecarboxylic acid derivatives (2006.01) Amine Compounds (6) Recrystallized solvent Yield (%) Melting point (℃)  8 N , N -dimethyl-4- ( 1H -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 4 Dimethylamine Ethanol Solution (33%) Methanol / ethyl acetate / hexanes (1: 5: 5) 38 164-166  9 N- [2- (dimethylamino) ethyl] -4- ( 1H -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 3 N, N -dimethylethylenediamine Chloroform / ether / methanol (3: 3: 1) 43 111-114  10 N- [2- (dimethylamino) ethyl] -4- ( 1H -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 4 N, N -dimethylethylenediamine Chloroform / ether / methanol (3: 3: 1) 35 162-167  11 4- ( 1H -5-indazolylamino) -2-methylthio- N- [2- (4-morpholinyl) ethyl] -5-pyrimidinecarboxamide Preparation Example 3 4- (2-aminoethyl) morpholine Chloroform / ether / methanol (3: 3: 1) 52 113-117  12 4- ( 1H -6-indazolylamino) -2-methylthio- N- [2- (4-morpholinyl) ethyl] -5-pyrimidinecarboxamide Preparation Example 4 4- (2-aminoethyl) morpholine Chloroform / ether / methanol (3: 3: 1) 56 226-228  13 4- (1 H-5-indazolyl-amino) - N - (4- methyl-1-piperazinyl) -2-methylthio-5-pyrimidine-carboxamide Preparation Example 3 1-amino-4-methylpiperazine Methylene chloride / hexane (1: 4) 64 251 to 253  14 4- (1 H -6- indazolyl amino) - N - (4- methyl-1-piperazinyl) -2-methylthio-5-pyrimidine-carboxamide Preparation Example 4 1-amino-4-methylpiperazine Methylene chloride / hexane (1: 4) 53 244-247  15 4- ( 1H -5-indazolylamino) -N -[( 1S ) -1- (methoxycarbonyl)-( 2R ) -2-hydroxypropyl] -2-methylthio-5-pyri Midinecarboxamide Preparation Example 3 L-Threonine Methyl Ester Methanol 76 227-232  16 4- ( 1H -6-indazolylamino) -N -[( 1S ) -1- (methoxycarbonyl)-( 2R ) -2-hydroxypropyl] -2-methylthio-5-pyri Midinecarboxamide Preparation Example 4 L-Threonine Methyl Ester Methanol 78 226-228  17 4- ( 1H -5-indazolylamino) -2-methylthio- N- (3-pyridyl) methyl-5-pyrimidinecarboxamide Preparation Example 3 3- (aminomethyl) pyridine Methanol / ether (1: 3) 62 268-270

Example number Name of the compound 5-pyrimidinecarboxylic acid derivatives (2006.01) Amine Compounds (6) Recrystallized solvent Yield (%) Melting point (℃)  18 4- ( 1H -6-indazolylamino) -2-methylthio- N- (3-pyridyl) methyl-5-pyrimidinecarboxamide Preparation Example 4 3- (aminomethyl) pyridine Methanol / ether (1: 3) 74 253-256  19 4- ( 1H -5-indazolylamino) -2-methylthio- N- [2- (2-pyridyl) ethyl] -5-pyrimidinecarboxamide Preparation Example 3 2- (2-aminoethyl) pyridine Methylene chloride / ether (1: 2) 49 220 to 223  20 4- ( 1H -6-indazolylamino) -2-methylthio- N- [2- (2-pyridyl) ethyl] -5-pyrimidinecarboxamide Preparation Example 4 2- (2-aminoethyl) pyridine Methylene chloride / ether (1: 2) 56 230-231  21 N- [3- ( 1H -1-imidazolyl) propyl] -4- ( 1H -5-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 3 1- (3-aminopropyl) imidazole Chloroform / isopropyl ether (1: 3) 53 121-125  22 N- [3- ( 1H -1-imidazolyl) propyl] -4- ( 1H -6-indazolylamino) -2-methylthio-5-pyrimidinecarboxamide Preparation Example 4 1- (3-aminopropyl) imidazole Methanol / ether (1: 3) 61 230-232  23 4- ( 1H -5-indazolylamino) -5-[(4-methyl-1-piperazinyl) carbonyl] -2-methylthiopyrimidine Preparation Example 3 1-methylpiperazine Methylene chloride / isopropyl ether (1: 2) 57 153-155  24 4- ( 1H -6-indazolylamino) -5-[(4-methyl-1-piperazinyl) carbonyl] -2-methylthiopyrimidine Preparation Example 4 1-methylpiperazine Methylene chloride / isopropyl ether (1: 2) 47 102-106  25 5-[[4- (2-hydroxyethyl) -1-piperazinyl] carbonyl] -4- (1 H -5-indazolylamino) -2-methylthiopyrimidine Preparation Example 3 1- (2-hydroxyethyl) piperazine Chloroform / isopropyl ether (1: 3) 58 166-170  26 5-[[4- (2-hydroxyethyl) -1-piperazinyl] carbonyl] -4- ( 1H -6-indazolylamino) -2-methylthiopyrimidine Preparation Example 4 1- (2-hydroxyethyl) piperazine Chloroform / isopropyl ether (1: 3) 52 114-117  27 4- ( 1H -5-indazolylamino) -2-methylthio- N- (4-morpholinyl) -5-pyrimidinecarboxamide Preparation Example 3 4-aminomorpholine Methanol / Water (1: 1) 60 239-242

Example NMR solvent ¹ H-NMR data (ppm) 2 DMSO-d ₆ δ 2.49 (s, 3H), 3.00 (s, 3H), 3.45 (br s, 2H), 3.56 (br s, 2H), 7.15 (d, 1H), 7.67 (d, 1H), 7.97 (s, 1H), 8.05 (br s, 1H), 8.22 (s, 1H), 9.04 (br s, 1H), 13.00 (br s, 1H) 3 DMSO-d ₆ δ0.88 (t, 3H), 1.45 (m, 1H), 1.63 (m, 1H), 2.47 (s, 3H), 3.45 (m, 2H), 3.89 (br s, 1H), 7.49 (m, 2H ), 8.03 (s, 1H), 8.12 (s, 1H), 8.32 (d, 1H), 8.71 (s, 1H), 11.22 (s, 1H), 13.03 (br s, 1H) 4 DMSO-d ₆ δ0.88 (t, 3H), 1.46 (m, 1H), 1.65 (m, 1H), 2.56 (s, 3H), 3.43 (m, 2H), 3.89 (br s, 1H), 7.05 (d, 1H ), 7.70 (d, 1H), 7.99 (s, 1H), 8.28 (s, 1H), 8.38 (d, 1H), 8.77 (s, 1H), 11.50 (s, 1H), 12.99 (br s, 1H ) 5 DMSO-d ₆ δ 2.46 (s, 3H), 3.28 (s, 3H), 3.46 (m, 4H), 7.45 (d, 1H), 7.52 (d, 1H), 8.03 (s, 1H), 8.12 (s, 1H) , 8.66 (s, 1H), 8.84 (br s, 1H), 11.22 (s, 1H), 13.05 (br s, 1H) 6 DMSO-d ₆ δ2.56 (s, 3H), 3.27 (s, 3H), 3.46 (m, 4H), 7.05 (d, 1H), 7.69 (d, 1H), 7.99 (s, 1H), 8.27 (s, 1H) , 8.71 (s, 1H), 8.89 (br s, 1H), 11.51 (br s, 1H), 13.05 (br s, 1H) 7 DMSO-d ₆ δ2.34 (s, 3H), 2.92 (s, 6H), 7.41 (m, 2H), 7.86 (d, 1H), 7.97 (s, 1H), 8.07 (d, 1H), 9.10 (s, 1H) , 12.96 (br s, 1 H), 8 DMSO-d ₆ δ 2.49 (s, 3H), 2.99 (s, 6H), 7.16 (d, 1H), 7.67 (d, 1H), 7.97 (s, 1H), 8.04 (d, 1H), 8.20 (s, 1H) , 9.31 (br s, 1 H), 13.00 (br s, 1 H) 9 DMSO-d ₆ δ 2.25 (s, 6H), 2.52 (m, 5H), 3.41 (m, 2H), 7.48 (d, 1H), 7.55 (d, 1H), 8.06 (s, 1H), 8.14 (s, 1H) , 8.65 (s, 1H), 8.76 (br s, 1H), 11.23 (br s, 1H), 13.10 (br s, 1H) 10 DMSO-d ₆ δ 2.18 (s, 6H), 2.42 (m, 2H), 2.49 (s, 3H), 3.36 (m, 2H), 7.45 (d, 1H), 7.52 (d, 1H), 8.03 (s, 1H) , 8.11 (s, 1H), 8.65 (s, 1H), 8.76 (br s, 1H), 11.22 (br s, 1H), 13.08 (br s, 1H) 11 DMSO-d ₆ + TFA-d ₁ δ2.50 (s, 3H), 3.14 (t, 2H), 3.34 (t, 2H), 3.54 (d, 2H), 3.68 (m, 4H), 3.96 (d, 2H), 7.48 (m, 1H) , 7.58 (d, 1 H), 8.06 (t, 2 H), 8.71 (s, 1 H) 12 DMSO-d ₆ δ 2.36 (s, 4H), 2.49 (s, 3H), 3.26 (s, 2H), 3.34 (d, 2H), 3.50 (s, 4H), 6.97 (d, 1H), 7.63 (d, 1H) , 7.91 (s, 1H), 8.20 (s, 1H), 8.61 (s, 1H), 8.71 (br s, 1H), 11.41 (s, 1H), 12.97 (br s, 1H) 13 DMSO-d ₆ + TFA-d ₁ δ 2.37 (s, 3H), 2.82 (s, 3H), 3.13-3.22 (m, 6H), 3.49 (d, 2H), 7.49 (d, 1H), 7.57 (d, 1H), 8.03 (s, 1H), 8.07 (s, 1H), 8.67 (s, 1H) 14 DMSO-d ₆ + TFA-d ₁ δ2.58 (s, 3H), 2.82 (s, 3H), 3.13-3.25 (m, 6H), 3.47 (d, 2H), 7.13 (d, 1H), 7.73 (d, 1H), 8.06 (s, 1H), 8.18 (s, 1H), 8.69 (s, 1H) 15 DMSO-d ₆ δ 1.16 (d, 3H), 2.47 (s, 3H), 3.68 (s, 3H), 4.19 (m, 1H), 4.50 (br s, 1H), 5.03 (d, 1H), 7.45 (d, 1H ), 7.52 (d, 1H), 8.04 (s, 1H), 8.10 (s, 1H), 8.71 (d, 1H), 8.80 (s, 1H), 10.86 (s, 1H), 13.06 (br s, 1H )  DMSO: dimethylsulfoxide, TFA: trifluoroacetic acid

Example NMR solvent ¹ H-NMR data (ppm) 16 DMSO-d ₆ δ 1.16 (d, 3H), 2.56 (s, 3H), 3.68 (s, 3H), 4.19 (m, 1H), 4.51 (br s, 1H), 5.04 (d, 1H), 7.05 (d, 1H ), 7.69 (d, 1H), 7.99 (s, 1H), 8.25 (s, 1H), 8.77 (d, 1H), 8.85 (s, 1H), 11.11 (br s, 1H), 13.05 (br s, 1H) 17 DMSO-d ₆ δ2.54 (s, 3H), 4.60 (d, 2H), 7.44 (m, 1H), 7.52 (d, 1H), 7.59 (d, 1H), 7.84 (d, 1H), 8.11 (d, 1H) , 8.19 (d, 1H), 8.54 (d, 1H), 8.66 (s, 1H), 8.78 (d, 1H), 9.43 (br s, 1H), 11.23 (s, 1H), 13.13 (br s, 1H ) 18 DMSO-d ₆ δ2.56 (s, 3H), 4.53 (d, 2H), 7.05 (m, 1H), 7.37 (m, 1H), 7.69 (d, 1H), 7.77 (m, 1H), 7.98 (s, 1H) , 8.27 (s, 1H), 8.47 (m, 1H), 8.59 (d, 1H), 8.77 (s, 1H), 9.41 (t, 1H), 11.43 (br s, 1H), 13.03 (br s, 1H ) 19 DMSO-d ₆ δ 2.47 (s, 3H), 3.01 (t, 2H), 3.63 (t, 2H), 7.22 (t, 1H), 7.29 (d, 1H), 7.45 (d, 1H), 7.52 (d, 1H) , 7.71 (t, 1H), 8.04 (s, 1H), 8.13 (s, 1H), 8.51 (s, 1H), 8.58 (s, 1H), 8.86 (br s, 1H), 11.17 (br s, 1H ), 13.06 (br s, 1H) 20 DMSO-d ₆ δ2.56 (s, 3H), 3.01 (t, 2H), 3.64 (t, 2H), 7.05 (d, 1H), 7.22 (t, 1H), 7.30 (d, 1H), 7.70 (d, 2H) , 7.99 (s, 1H), 8.27 (s, 1H), 8.50 (s, 1H), 8.63 (s, 1H), 8.92 (br s, 1H), 11.47 (br s, 1H), 13.03 (br s, 1H) 21 DMSO-d ₆ δ 1.98 (t, 2H), 2.47 (s, 3H), 3.24 (t, 2H), 4.04 (t, 2H), 6.88 (s, 1H), 7.21 (s, 1H), 7.45 (d, 1H) , 7.52 (d, 1H), 7.66 (s, 1H), 8.03 (s, 1H), 8.13 (s, 1H), 8.65 (s, 1H), 8.76 (br s, 1H), 11.12 (br s, 1H ), 13.05 (br s, 1H) 22 DMSO-d ₆ δ 1.98 (m, 2H), 2.57 (s, 3H), 3.26 (m, 2H), 4.04 (t, 2H), 6.89 (s, 1H), 7.05 (d, 1H), 7.21 (s, 1H) , 7.69 (t, 2H), 7.98 (s, 1H), 8.28 (s, 1H), 8.69 (s, 1H), 8.81 (t, 1H), 11.43 (br s, 1H), 13.06 (br s, 1H ) 23 DMSO-d ₆ + TFA-d ₁ δ 2.48 (s, 3H), 2.82 (s, 3H), 3.06 (t, 2H), 3.31 (br s, 2H), 3.47 (d, 2H), 4.26 (br s, 2H), 7.45 (d, 1H), 7.56 (d, 1H), 7.87 (s, 1H), 8.06 (s, 1H), 8.31 (s, 1H) 24 DMSO-d ₆ + TFA-d ₁ δ 2.44 (s, 3H), 2.77 (s, 3H), 3.00 (t, 2H), 3.25 (br s, 2H), 3.42 (d, 2H), 4.24 (br s, 2H), 7.18 (d, 1H), 7.69 (d, 1H), 7.86 (s, 1H), 8.00 (s, 1H), 8.30 (s, 1H) 25 DMSO-d ₆ δ 2.39 (t, 5H), 2.49 (d, 4H), 3.48 (m, 6H), 4.13 (t, 1H), 7.44 (d, 1H), 7.49 (d, 1H), 7.92 (d, 1H) , 8.02 (s, 1H), 8.08 (s, 1H), 9.11 (s, 1H), 13.02 (br s, 1H) 26 DMSO-d ₆ + TFA-d ₁ δ 2.45 (s, 3H), 3.14 (d, 4H), 3.49 (br s, 4H), 3.72 (s, 2H), 4.22 (br s, 2H), 7.22 (d, 1H), 7.71 (d, 1H), 7.89 (s, 1H), 8.05 (s, 1H), 8.34 (s, 1H) 27 DMSO-d ₆ δ2.53 (s, 3H), 2.96 (s, 4H), 3.74 (s, 4H), 7.52 (d, 1H), 7.59 (d, 1H), 8.06 (s, 1H), 8.19 (s, 1H) , 8.61 (s, 1H), 9.83 (s, 1H), 10.95 (s, 1H), 13.12 (br s, 1H)

Experimental Example 1 Inhibitory Effect of In Vitro Reverse Transcription Activity on HBV Polymerase

In order to determine the effect of the compounds of Formula 1 inhibit the reverse transcription activity of HBV polymerase, the following in vitro experiment was performed.

The present inventors have already filed a patent for a recombinant polymerase protein of HBV expressed in E. coli, a method for preparing the same, and a method for measuring the enzyme activity thereof (Korean Patent Application Nos. 94-3918 and 96-33998). In this experiment, HBV polymerase expressed in E. coli was used as described above.

The method for measuring reverse transcriptase activity of hepatitis B virus polymerase in vitro used in the present invention is as follows. The basic principle is the same as the enzyme-immunological method (ELISA), in which the nucleotides modified with biotin- and DIG- (digoxigenin-) are reacted with the substrate, and then the polymerized substrate with the anti-peroxidase The method of recognizing with -DIG antibody was used.

20 μl of HBV polymerase was added to a well coated with streptavidin and the reaction mixture [10 μM each of DIG-UTP, Biotin-UTP, 46 mM Tris-HCl, 266 mM KCl, 27.5 mM MgCl ₂ , 9.2 mM DTT substrate / primer hybrid] 20 µl and 20 µl of the test substance (addition of 1, 0.1 and 0.01 µg / ml, respectively) were mixed and reacted at 22 ° C for 15 hours. The DNA is produced by the action of HBV polymerase and contains digoxigenin and biotin-attached nucleotides, which binds to streptavidin coated on the bottom of the well. After the reaction was completed, washed 5 times 30 seconds with 250 μl of wash buffer (pH 7.0) per well to remove the remaining impurities. After 200 μl of anti-DIG-POD antibody was added to each well and reacted at 37 ° C. for 1 hour, each well was washed with a washing buffer to remove impurities. Thereafter, 200 μl of ABTS ™, a substrate of POD (peroxidase), was added thereto, and the mixture was reacted at room temperature for 30 minutes, and the absorbance at 405 nm was measured using an ELISA reader.

Inhibition rate of the reverse transcription activity of the HBV polymerase was calculated based on the control without the test compound, the results are shown in Table 3 below.

Inhibitory Effect on Reverse Transcriptase Activity of HBV Polymerase Test compound % Inhibition of activity of HBV-RT 1 μg / ml 0.1 μg / ml 0.01 μg / ml Example 1 75 54 30 Example 2 70 48 22 Example 3 82 59 44 Example 4 85 55 36 Example 5 58 40 13 Example 6 60 42 25 Example 7 77 61 42 Example 8 54 30 10 Example 9 80 51 15 Example 10 92 66 54 Example 11 64 39 10 Example 12 67 40 12 Example 13 78 54 40 Example 14 72 46 28 Example 15 51 40 18 Example 16 82 63 50 Example 17 97 76 53 Example 18 98 79 55 Example 19 88 60 48

Inhibitory Effect on Reverse Transcriptase Activity of HBV Polymerase Test compound % Inhibition of activity of HBV-RT 1 μg / ml 0.1 μg / ml 0.01 μg / ml Example 20 95 70 52 Example 21 80 56 41 Example 22 91 58 32 Example 23 82 58 43 Example 24 62 40 26 Example 25 53 32 11 Example 26 59 43 15 Example 27 90 61 46

As can be seen in Table 3 , the compounds of the present invention have a very good effect of inhibiting the activity of HBV polymerase, such as at least 70%, up to 98% of the activity inhibition rate for HBV polymerase at a concentration of 1 ㎍ / ㎖ Do. In addition, since the compounds of the present invention are non-nucleic acid-based materials, it is expected to solve problems such as the early appearance of toxic and resistant viruses possessed by nucleic acid-based materials, and they may be used in combination with nucleic acid-based materials because they have different mechanisms of action. It can also be used as a therapy.

As described above, the compounds of the present invention have an excellent effect of inhibiting the activity of HBV polymerase, which plays an important role in the replication of HBV. Therefore, the compounds of the present invention can inhibit the proliferation of HBV. Can be used.

Experimental Example 2 Effect of HBV Proliferation Inhibitory Activity Using HBV Producing Cell Line

In order to determine the effects of the compounds of Formula 1 inhibit the proliferation of HBV producing cell line, the following experiment was carried out.

To detect antiviral activity, HepG 2.2.15, a human liver cancer cell line, was used to measure the extent of HBV replication and proliferation.

The cell concentration was adjusted to 1 × 10 ⁵ cells / ml and then dispensed in 1 ml / well into 24-well cell culture plates. This was incubated in a 5% CO ₂ incubator at 37 ° C., the medium was changed daily and incubated for 3-4 days until the cells were sufficiently grown (confluent). After sufficient growth of the cells, test compounds were added so that the final concentrations were 0.01, 0.1, and 1 μg / ml, respectively. One week after the addition of the test compound, the culture medium was taken and centrifuged at 5,000 rpm for 10 minutes. 25 μl of supernatant was transferred to a new tube, and 5 μl of lysis solution [0.54N NaOH, 0.06% NP40] was added and incubated at 37 ° C. for 1 hour. After incubation, 30 μl of neutralization solution [0.09N HCl, 0.1M Tris-HCl, pH 7.4] was added and used as a reaction solution for competitive PCR (Competitive Polymerase Chain Reaction).

PCR was performed using the core gene sequence of HBV as a template. 1 unit of Taq polymerase was added to 25 pmol of each primer, 250 μM dNTP, 5 μl of the above PCR reaction solution [0.54N NaOH, 0.06% NP40, 0.09N HCl, 0.1M Tris-HCl, pH 7.4]. PCR reaction was performed.

DNA amplified by PCR was subjected to agarose gel electrophoresis, and then quantitative analysis of HBV DNA using an image analyzer (Gel Doc 1000, BIO-RAD) to evaluate the HBV growth inhibitory activity of the compounds of the present invention.

3TC (lamivudine) was used as a positive control and treated at the same concentration as the test compound. HBV proliferation inhibitory activity was calculated based on the control without the test compound, the results are shown in Table 4 below.

HBV proliferation inhibitory activity Test compound HBV proliferation inhibitory activity rate (%) 1 μg / ml 0.1 μg / ml 0.01 μg / ml Example 1 65 48 20 Example 7 70 52 25 Example 10 88 57 43 Example 17 92 65 40 Example 18 97 75 47 Example 19 84 55 38 Example 20 93 68 44 Example 27 86 50 29 3TC 99 80 48

As can be seen in Table 4 , the compounds of the present invention as a non-nucleic acid-based substance inhibits the reverse transcription activity of HBV polymerase, such as at least 80%, up to 97% proliferation inhibitory activity against HBV at a concentration of 1 ㎍ / ㎖ The effect is very good. In particular, since the compounds of the present invention are non-nucleic acid-based materials, it is expected to solve problems such as the early appearance of toxic and resistant viruses possessed by nucleic acid-based materials. In addition, since the nucleic acid-based substances act on the active domain of the polymerase, the compounds of the present invention are expected to act on the allosteric binding pocket, so the compounds of the present invention may be used as a combination therapy with nucleic acid-based substances. Can be used.

As such, the compounds of the present invention have an excellent effect of inhibiting the reverse transcriptase activity of HBV polymerase, which plays an important role in the reverse transcription phase of HBV replication. And as therapeutic agents.

Experimental Example 3 In Vitro Inhibitory Effect on HIV Reverse Transcriptase

In order to determine the effect of the compounds of Formula 1 inhibit the activity of HIV reverse transcriptase, the following in vitro experiments were carried out.

In vitro inhibitory activity was measured using a non-radioactive reverse transcriptase assay kit (Boehringer Mannheim).

20 μl (40 ng) of HIV reverse transcriptase was added to a well coated with streptavidin, and the template- primer hybrid poly (A) oligo (dT) ₁₅ and DIG- (digoxigenin) -dUTP, biotin- After 20 µl of a reaction mixture containing dUTP and TTP was added, the test compound was added thereto to a final concentration of 0.1 and 1 µg / ml, and reacted at 37 ° C for 1 hour. At this time, DNA is made from RNA by the action of HIV reverse transcriptase, and since digoxigenin and biotin are labeled together, streptavidin is coated on the bottom of the well. Will be combined.

After completion of the reaction, washing was performed 5 times for 30 seconds with 250 μl of washing buffer solution (pH 7.0) per well to remove remaining impurities. After washing, 200 μl of anti-DIG-POD antibody was added thereto and reacted at 37 ° C. for 1 hour, and the resultant was washed with the same buffer solution for washing to remove impurities. After washing, 200 μl of ABTS ™, a substrate of POD (peroxidase), was added to each well and allowed to react at room temperature for 30 minutes. After completion of the reaction, the degree of inhibition of reverse transcriptase activity of HIV was quantified by measuring the absorbance at 405 nm of each solution using an ELISA reader. Inhibition rate for HIV reverse transcriptase activity was calculated based on the control without the test compound, the results are shown in Table 5 below.

Inhibitory Effect on HIV Reverse Transcriptase Activity Test compound HIV-RT activity inhibition rate (%) 1 μg / ml 0.1 μg / ml Example 1 84 52 Example 2 80 47 Example 8 81 53 Example 10 77 51 Example 11 75 48 Example 12 89 50 Example 21 72 44

As can be seen in Table 5 , the compounds of the present invention inhibit the activity of HIV reverse transcriptase, such as at least 80%, up to 89% of the activity inhibition against HIV-RT (HIV reverse transcriptase) at a concentration of 1 ㎍ / ㎖ The effect is very good. In particular, since the compounds of the present invention are non-nucleic acid-based materials, it is expected to solve problems such as the early appearance of toxic and resistant viruses possessed by nucleic acid-based materials. In addition, since the nucleic acid-based substances act on the active domain of the polymerase, the compounds of the present invention are expected to act on the allosteric binding pocket, so the compounds of the present invention may be used as a combination therapy with nucleic acid-based substances. Can be used.

As described above, the compounds of the present invention have an excellent effect of inhibiting the activity of HIV reverse transcriptase, which plays an important role in the reverse transcription phase of HIV replication. Therefore, the compounds of the present invention can suppress the proliferation of HIV, thus preventing and preventing AIDS. It can be usefully used as a therapeutic agent.

Experimental Example 4 Cytotoxicity Test

In order to determine whether the compounds of Formula 1 exhibit cytotoxicity, in vitro experiments were carried out using HepG2 cells in a generally known MTT assay method.

As a result, all of the compounds used in the experiment were found to have a CC ₅₀ of 100 µg / ml or more, which is a very low toxicity to cells.

Experimental Example 5 Oral Acute Toxicity in Rats

In order to determine the acute toxicity of the compound of Formula 1 was performed the following experiment.

Acute toxicity experiments were performed using 6-week-old SPF SD rats. Six animals per group were suspended orally at a dose of 4 g / kg / 15 ml, each of the compounds of Examples 1-27 suspended in 0.5% methylcellulose solution. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and blood biochemical tests were performed. Necropsy was performed to visually check for abdominal and thoracic organ abnormalities. As a result, all animals treated with test substance showed no clinical symptoms and no dead animals, and no toxic changes were observed in weight change, blood test, blood biochemistry, autopsy findings. As a result, all of the tested compounds did not show toxic changes up to 4 g / kg in rats, and the minimum lethal dose (LD ₅₀ ) for oral administration was determined to be a safe substance of 4 g / kg or more.

As described above, the novel 5-pyrimidinecarboxamide derivative represented by Chemical Formula 1 according to the present invention is excellent in inhibiting the proliferation of HBV and HIV and has fewer side effects, so hepatitis B and acquired immunodeficiency syndrome. It can be usefully used as a prophylactic and therapeutic agent for. In particular, since the compounds of the present invention are non-nucleic acid-based materials, it is expected to solve problems such as early appearance of toxic and resistant viruses possessed by nucleic acid-based materials. In addition, since the nucleic acid compounds act on the active domain of the polymerase while the compounds of the present invention are expected to act on the allosteric binding pocket, the compounds of the present invention may be used as a combination therapy with nucleic acid compounds. have.

Claims (4)

5-pyrimidinecarboxamide derivatives represented by Formula 1 or a pharmaceutically acceptable salt thereof: Formula 1

(In Formula 1, R ₁ is hydroxy; C ₁ -C ₅ linear or pulverized alkyl group; C ₁ -C ₅ linear or pulverized alkoxy group; C ₂ -C ₆ linear or pulverized hydroxyalkyl group; C ₂ -C ₆ dialkylamino group; C ₂ ~C ₅ is an alkoxycarbonyl group and a hydroxyl substituted with either one or both of the time, C ₂ ~C ₆ is a straight or branched chain alkyl group; Or is optionally substituted with C ₁ ~C ₃ alkyl group, and a N, O, S, a saturated or unsaturated 5-atom or 6-atom heterocyclic compound containing 1 to 3 heteroatoms selected from substituted, R ₁ is asymmetrical With or without carbon, R ₂ is H; Or a C ₁ -C ₄ straight or crushed alkyl group, Or R ₁ and R ₂ combine with each other to form a saturated 5- or 6-membered heterocyclic compound, wherein the heterocycle contains 1 to 3 heteroatoms selected from N, O, and S and the heterocycle is unsubstituted. Or substituted with a C ₁ -C ₅ straight or crushed alkyl group or C ₂ -C ₅ straight or pulverized hydroxyalkyl group, R ₃ is indazol-5-yl; Or indazol-6-day, n is an integer of any one of 0 to 4, With the proviso that when n is 0 and R ₂ is H, R ₁ is an alkyl group of C ₁ to C ₄ . 1) 4-chloro-2-methylthio-5-pyrimidinecarboxylic acid ethyl ester ( 2 ) and 5-aminoindazole or 6-aminoindazole ( 3 ) with triethylamine, N, N -diisopropyl Reacted in the presence of one base selected from the group consisting of ethylamine, N -methylmorpholine, N -methylpiperidine, 4-dimethylaminopyridine, N, N -dimethylaniline, 2,6-lutidine and pyridine Preparing 5-pyrimidinecarboxylic acid ethyl ester derivative ( 4 ) (step 1); 2) preparing a 5-pyrimidinecarboxylic acid derivative ( 5 ) by hydrolyzing the 5-pyrimidinecarboxylic acid ethyl ester derivative ( 4 ) prepared in step 1 under alkaline conditions (step 2); And 3) the 5-pyrimidinecarboxylic acid derivative ( 5 ) prepared in step 2 was activated with a Vilsmeier intermediate in the presence of N, N -dimethylformamide (DMF) and thionyl chloride (SOCl ₂ ). To prepare a 5-pyrimidinecarboxamide derivative ( 7 ) by reacting with an amine compound ( 6 ) (step 3). Scheme 1

(In Scheme 1, R ₁ is hydroxy; C ₁ -C ₅ linear or pulverized alkyl group; C ₁ -C ₅ linear or pulverized alkoxy group; C ₂ -C ₆ linear or pulverized hydroxyalkyl group; C ₂ -C ₆ dialkylamino group; C ₂ ~C ₅ is an alkoxycarbonyl group and a hydroxyl substituted with either one or both of the time, C ₂ ~C ₆ is a straight or branched chain alkyl group; Or is optionally substituted with C ₁ ~C ₃ alkyl group, and a N, O, S, a saturated or unsaturated 5-atom or 6-atom heterocyclic compound containing 1 to 3 heteroatoms selected from substituted, R ₁ is asymmetrical With or without carbon, R ₂ is H; Or a C ₁ -C ₄ straight or crushed alkyl group, Or R ₁ and R ₂ combine with each other to form a saturated 5- or 6-membered heterocyclic compound, wherein the heterocycle contains 1 to 3 heteroatoms selected from N, O, and S and the heterocycle is unsubstituted. Or substituted with a C ₁ -C ₅ straight or crushed alkyl group or C ₂ -C ₅ straight or pulverized hydroxyalkyl group, R ₃ is indazol-5-yl; Or indazol-6-day, n is an integer of any one of 0 to 4). A pharmaceutical composition for treating or preventing hepatitis B, comprising the 5-pyrimidinecarboxamide derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for treating or preventing AIDS, comprising the 5-pyrimidinecarboxamide derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.