KR20000020792A - Novel 2,5-pyridine carboxylic derivatives - Google Patents

Abstract

PURPOSE: Title derivatives are provided which has an inhibition effect against HBV(Hepatitis B Virus) and HIV(Human Immunodeficiency Virus) growth. CONSTITUTION: Nicotinic derivatives(formula 2; R4 is H or C1-C4 straight chain or branched alkyl group) and amine compound(formula 3; R1 is C1-C4 straight chain or branched alkoxy or amino group; R2 is H, C1-C4 straight chain or branched alkyl, C1-C3 straight chain or branched hydroxy alkyl, C2-C4 alkyl thioalkyl, phenyl, benzyl or hydroxy benzyl group; R3 is H or methyl group; n is 0-1) are reacted to give the derivatives(formula 1). Thus, 2 g of 6-£1-£3-isopropyl amino)-2-pyridyl|piperazin-4-yl-carbonyl|nicotine acid and 0.8 ml of triethyl amine are dissolved in 30 ml of methylene chloride, and slowly added 0.7 ml of pivaloyl chloride at 0- 5°C. Reactant is reacted at 5°C for 1 hour, followed by addition of 2.5 ml of N,N-diisopropyl ethyl amine and 0.83 g of glycine ethyl ester at 10°C for 2 hour to give 1.89 g of N-(ethoxycarbonyl methyl)-6-£1-£3-isopropyl amino)-2-pyridyl|piperazin-4-yl-carbonyl|nicotine amide hydrochloride.

Description

New 2,5-pyridinedicarboxylic acid derivatives

The present invention relates to a novel 2,5-pyridinedicarboxylic acid derivative, and more particularly, to Formula 1, which is effective as an antiviral agent due to its excellent anti-proliferative effect against Hepatitis B Virus (HBV) and Human Immunodeficiency Virus (HIV). Novel 2,5-pyridinedicarboxylic acid derivatives, pharmaceutically acceptable salts thereof, and methods for preparing the same are disclosed.

Formula 1

In Formula 1, R ₁ represents a linear or pulverized alkoxy group, a hydroxy group or an amino group having 1 to 4 carbon atoms, and R ₂ represents a hydrogen atom, a linear or pulverized alkyl group having 1 to 4 carbon atoms, a carbon source When a linear or pulverized hydroxyalkyl group of embroidery 1 to 3, an alkylthioalkyl group having 2 to 4 carbon atoms, a phenyl group, a benzyl group or a hydroxybenzyl group is represented, and R ₂ represents a substituent other than a hydrogen atom, (R) -Or (S)-represents stereospecificity, R ₃ represents a hydrogen atom or a methyl group, R ₄ represents a hydrogen atom or a linear or pulverized alkyl group having 1 to 4 carbon atoms, n is 0 or 1 .

Hepatitis B virus (HBV) is a major pathogen of hepatitis with an infection of about 300 million people worldwide. It not only invades humans and causes acute or chronic hepatitis, but also cirrhosis or liver cancer. It is well known to be involved in the transition. Much research has been conducted on HBV due to its association with liver disease and molecular biological characteristics of the virus. However, only preventive vaccines for hepatitis B have been developed so far, and no obvious treatment has been developed. Some of the Nucleosides compounds, which have been developed primarily for the treatment of Acquired immune deficiency syndrome (AIDS), have been reported to be effective as HBV inhibitors for some time [J. Org. Chem., 1992, Vol. 57, 2217]. These nucleic acid-based compounds are known to be limited to the development of a desirable hepatitis B therapeutic agent due to the high cost of treatment and inherent side effects and toxicity of the nucleic acid-based compounds.

Therefore, there is an urgent need for development of a hepatitis B therapeutic agent having excellent antiviral action against HBV as a non-nucleic acid compound, and recently, a quinolone compound [European Patent Publication Nos. Korean Patent Publication No. 94-1886] and the like have been applied for a patent, but until now, no clear development result is known.

The present inventors have conducted continuous research to solve side effects and toxicity problems and to develop non-nucleic acid compounds having excellent antiviral activity against HBV. As a result, three inventions and new 2,5-pyridinedicarboxylic acid are described. We have already filed three inventions on derivatives, one invention on 4-substituted piperidinoterephthalic acid derivatives, and one invention on novel (2-hydroxyethyl) piperazine derivatives (Korean Patent Application No. 96- 72384, 97-36589, 97-61308, 97-77830, 98-12218, 98-17153, 98-19555 and 98-28258. In addition, the novel 2,5-pyridinedicarboxylic acid derivative represented by Formula 1 of the present invention is a non-nucleosides having a unique structure as well as HBV proliferation inhibitory effect as well as human immunodeficiency virus (Human Immunodeficiency virus, hereinafter) It was confirmed that the proliferation inhibitory effect on the "HIV") was completed the present invention. For example, HBV and HIV are different viruses, but they have a common replication process, namely reverse transcription from viral RNA to DNA, and the cleavage of the RNA portion of the resulting RNA-DNA hybrid. , 2,5-pyridinedicarboxylic acid derivative represented by the formula (1) of the present invention has a mechanism of action that inhibits this process can be developed as a growth inhibitor for HBV and HIV.

Accordingly, an object of the present invention is to provide a novel 2,5-pyridinedicarboxylic acid derivative having a strong anti-proliferative effect on HBV and HIV, a pharmaceutically acceptable salt thereof, and a preparation method thereof.

The present invention is characterized by a novel 2,5-pyridinedicarboxylic acid derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof.

Formula 1

In Formula 1, R ₁ represents a linear or pulverized alkoxy group, a hydroxy group or an amino group having 1 to 4 carbon atoms, and R ₂ represents a hydrogen atom, a linear or pulverized alkyl group having 1 to 4 carbon atoms, a carbon source When a linear or pulverized hydroxyalkyl group of embroidery 1 to 3, an alkylthioalkyl group having 2 to 4 carbon atoms, a phenyl group, a benzyl group or a hydroxybenzyl group is represented, and R ₂ represents a substituent other than a hydrogen atom, (R) -Or (S)-represents stereospecificity, R ₃ represents a hydrogen atom or a methyl group, R ₄ represents a hydrogen atom or a linear or pulverized alkyl group having 1 to 4 carbon atoms, n is 0 or 1 .

Referring to the present invention in more detail as follows.

In the 2,5-pyridine dicarboxylic acid derivative represented by Chemical Formula 1 according to the present invention, preferably, R ₁ represents a methoxy group, an ethoxy group, a hydroxy group or an amino group, and R ₂ represents a hydrogen atom, a methyl group or an iso Propyl group, isobutyl group, hydroxymethyl group, 1-hydroxyethyl group, (2-methylthio) ethyl group, (2-ethylthio) ethyl group, phenyl group, benzyl group, 4-hydroxybenzyl group, wherein R ₃ is Represents a hydrogen atom or a methyl group, and R ₄ represents a hydrogen atom, an ethyl group, an isopropyl group or an isobutyl group. When R ₂ represents a substituent other than a hydrogen atom, stereospecificity in the form of (R)-or (S)- Indicates.

Representative examples of the 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 according to the present invention are as follows.

N- (ethoxycarbonylmethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (carboxymethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1- (ethoxycarbonyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1-carboxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1- (ethoxycarbonyl) -2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbo Nil] nicotinamide

N-[(1S) -1-carboxy-2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1R) -1- (ethoxycarbonyl) -2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbo Nil] nicotinamide

N-[(1S, 2R) -2-hydroxy-1- (methoxycarbonyl) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl -Carbonyl] nicotinamide

N-[(1S, 2R) -1-carboxy-2-hydroxypropyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotin amides

N-[(1S) -1- (ethoxycarbonyl) -3- (methylthio) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl -Carbonyl] nicotinamide

N-[(1S) -1-carboxy-3- (methylthio) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotin amides

N-[(1S) -1- (ethoxycarbonyl) -2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine- 4-yl-carbonyl] nicotinamide

N-[(1S) -1-carboxy-2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbo Nil] nicotinamide

N- (carbamoylmethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1-carbamoyl-2-methylpropyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1-carbamoyl-2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl- Carbonyl] nicotinamide

N- (1-carboxy-3-hydroxypropyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (ethoxycarbonylmethyl) -N-methyl-6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (carboxymethyl) -N-methyl-6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (ethoxycarbonylethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (carboxyethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

In addition, the 2,5-pyridinedicarboxylic acid derivative represented by Formula 1 according to the present invention may form a pharmaceutically acceptable acid addition salt according to a conventional method in the art. Examples include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or formic acid, acetic acid, propionic acid, oxalic acid, citric acid, maleic acid, succinic acid, fumaric acid, tartaric acid, trifluoroacetic acid, benzoic acid, methanesulfonic acid, and the like. Salts with organic acids.

The 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 according to the present invention has a strong antiproliferative effect on HBV and HIV and thus may be used as a clinically useful hepatitis B therapeutic agent and AIDS therapeutic agent. Accordingly, the present invention includes a pharmaceutical composition containing the 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 and a pharmaceutically acceptable salt thereof as an active ingredient.

In clinical use of the pharmaceutical composition of the present invention, it is combined with a conventional carrier in the pharmaceutical field, and is a conventional agent in the pharmaceutical field, for example, an oral preparation such as tablets, capsules, troches, solutions, suspensions, and the like. ; 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.

Pharmaceutical formulations prepared using conventional carriers can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically.

In addition, the dosage of the 2,5-pyridinedicarboxylic acid derivative represented by Formula 1 according to the present invention to the human body is generally administered to an adult in an amount of 10 to 500 mg, preferably 50 to 300 mg per day. In accordance with the judgment of the doctor or pharmacist, it may be divided several times a day, preferably once or six times, at regular time intervals.

The present invention includes a method for preparing a 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1, wherein the 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 according to the present invention is prepared by the following Scheme 1. can do.

The following Reaction Scheme 1 briefly illustrates a method of preparing a 2,5-pyridinedicarboxylic acid derivative represented by Formula 1 as a target compound by reacting a nicotinic acid derivative represented by Formula 2 with an amine compound represented by Formula 3 below:

In Reaction Scheme 1, R ₁ represents a linear or pulverized alkoxy group having 1 to 4 carbon atoms or an amino group, and R ₂ , R ₃ , R ₄ and n are as defined above.

Hereinafter, the preparation process of the 2,5-pyridinedicarboxylic acid derivative according to the present invention as shown in Scheme 1 will be described in more detail.

The nicotinic acid derivative represented by Chemical Formula 2 is reacted with an acid chloride such as pivaloyl chloride to generate a highly reactive acid anhydride or by reacting with thionyl chloride to generate a highly reactive acid chloride. The 2,5-pyridine dicarboxylic acid derivative represented by the formula (1) as a target compound is obtained by reacting with an amine compound represented by the formula (1). These reactions are triethylamine, N, N-diisopropylethylamine, N-methylmorpholine, N-methylpiperidine, 4-dimethylaminopyridine, N, N-dimethylaniline, 2,6-lutidine, In the presence of a general tertiary organic base such as pyridine or the like, the amine compound represented by Chemical Formula 3 may be used in an amount of 1 to 1.5 equivalents or the amine compound represented by Chemical Formula 3 may be used in an excessive amount without using a tertiary organic base. It may be. Alternatively, the nicotinic acid derivative represented by Chemical Formula 2 is reacted with 1-hydroxybenzotriazole (HOBT) in the presence of 1,3-dicyclohexylcarbodiimide (DCC) to generate an active ester having high reactivity. It is reacted with the amine compound represented by 3, and the 2, 5- pyridine dicarboxylic acid derivative represented by the said General formula (1) which is a target compound is obtained. This reaction is completed within 8 hours at 5 ° C to 40 ° C. Solvents used in the reactions include chloroform, methylene chloride, acetonitrile, tetrahydrofuran, dioxane, dioxolane, N, N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, and the like. It is preferable to use a single solvent or a mixed solvent selected.

In addition, the preparation method of the nicotinic acid derivative represented by Formula 2 used as a starting material in the preparation method of the compound represented by Formula 1 according to Scheme 1 is described in Korean Patent Application Nos. 97-61308 and 98-17153 It can be manufactured and used according to the method.

The amine compound represented by Chemical Formula 3 used in Scheme 1 is a substituent of the 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 As a material for introducing a, it is selected from a suitable amine compound commercially available according to the required substituent, which can be easily selected and used by those of ordinary skill in the art.

When R ₁ is an organic acid derivative representing a hydroxy group in the target compound represented by Formula 1, an ester derivative in which R ₁ represents a methoxy or ethoxy group in the compound represented by Formula 1 prepared in Scheme 1 It can be easily prepared by hydrolysis by the method.

Such a present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1 N- (ethoxycarbonylmethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide hydrochloride

To methylene chloride (30 mL) add 6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinic acid (2 g) and triethylamine (0.8 mL) It was dissolved by addition. After cooling, pivaloyl chloride (0.7 mL) was slowly added at 0 ° C to 5 ° C, and reacted at 5 ° C for 1 hour. N, N-diisopropylethylamine (2.5 ml) and glycine ethyl ester hydrochloride (0.83 g) were added and reacted at 10 ° C. for 2 hours.

The reaction solution was washed with aqueous sodium bicarbonate solution and water, and the separated organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The concentrated residue was purified by column chromatography (chloroform: isopropanol: hexane = 5: 1: 1). The purified solvent layer was recovered, concentrated under reduced pressure, and the residue was dissolved in methylene chloride (20 ml), 2 ml of anhydrous hydrochloric acid (2N-ethanol solution) was added thereto, concentrated under reduced pressure, crystallized by treating with ether and filtered. Yield 71%) of the title compound was prepared as crystals of hydrochloride.

¹ H-NMR (CDCl ₃ ), ppm: δ 1.29 (m, 9H), 3.56 (m, 5H), 3.86 (m, 2H), 4.03 (m, 2H), 4.27 (m, 4H), 7.23 ( m, 2H), 7.75 (m, 1H), 7.81 (m, 1H), 8.31 (m, 1H), 9.03 (s, 1H)

Example 2 N- (carboxymethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (ethoxycarbonylmethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl obtained in Example 1 above in methanol (20 mL) ] Ninicotinamide hydrochloride (1.5 g) was dissolved, 1N aqueous sodium hydroxide solution (10 ml) was added, and the mixture was reacted at 30 占 폚 to 40 占 폚 for 2 hours. At 20 ° C., 1N hydrochloric acid was gradually added to neutralize (pH-5), and excess water was gradually added to precipitate. After stirring for 2 hours, the mixture was filtered, washed with water and dried to obtain 1.17 g (yield 90%) of the title compound.

m.p. : 161-164 ℃

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.45 (d, 6H), 3.70 (m, 4H), 3.88 (m, 1H), 4.10 (m, 2H), 4.34 (m, 2H), 4.62 (m, 2H), 7.58 (m, 1H), 7.74 (m, 2H), 8.58 (d, 1H), 9.38 (d, 1H), 9.74 (s, 1H)

Example 3 N-[(1S) -1- (ethoxycarbonyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl ] Nicotinamide

To methylene chloride (30 mL) add 6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinic acid (2 g) and triethylamine (0.8 mL) It was dissolved by addition. After cooling, pivaloyl chloride (0.7 mL) was slowly added at 0 ° C to 5 ° C, and reacted at 5 ° C for 1 hour. Triethylamine (2.3 mL) and L-alanine ethyl ester hydrochloride (0.92 g) were added thereto, followed by reaction at 5 ° C to 10 ° C for 2 hours, and then gradually heated to further react at 15 ° C to 20 ° C for 30 minutes.

The reaction solution was washed with aqueous sodium bicarbonate solution and water, and the separated organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The concentrated residue was purified by column chromatography (ethyl acetate: isopropanol: hexane = 5: 2: 1). The solvent layer was recovered, concentrated under reduced pressure, and the residue was treated with ether to crystallize, stirred for 1 hour, and filtered to obtain 1.93 g (yield 76%) of the title compound.

¹ H-NMR (CDCl ₃ ), ppm: δ 1.24 (d, 6H), 1.32 (t, 3H), 1.55 (d, 3H), 3.10 (m, 2H), 3.19 (m, 2H), 3.53 ( m, 1H), 3.71 (m, 2H), 3.97 (m, 2H), 4.17 (m, 1H), 4.27 (m, 2H), 4.79 (m, 1H), 6.86 (m, 1H), 6.94 (m , 1H), 7.04 (m, 1H), 7.68 (m, 2H), 8.21 (m, 1H), 9.00 (s, 1H)

Example 4 N-[(1S) -1-carboxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N-[(1S) -1- (ethoxycarbonyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl- obtained in Example 3 above Carbonyl] nicotinamide was hydrolyzed by the method of Example 2 to prepare a target compound.

Yield: 93%

m.p. : 240 ~ 245 ℃

¹ H-NMR (DMSO-d ₆ + TFA-d ₁ ), ppm: δ 0.96 (d, 6H), 1.29 (d, 3H), 3.21 (m, 4H), 3.41 (m, 1H), 3.51 ( m, 2H), 3.80 (m, 2H), 4.50 (m, 1H), 7.07 (m, 1H), 7.16 (m, 1H), 7.29 (m, 1H), 7.98 (m, 1H), 8.81 (m , 1H), 9.19 (s, 1H)

Example 5 N-[(1S) -1- (ethoxycarbonyl) -2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4 -Yl-carbonyl] nicotinamide

L-serine ethyl ester hydrochloride was carried out in the same manner as in Example 3 to prepare a target compound.

Yield: 67%

¹ H-NMR (CDCl ₃ ), ppm: δ 1.24 (d, 6H), 1.33 (t, 3H), 3.32 (m, 4H), 3.56 (m, 3H), 3.95 (m, 2H), 4.12 ( m, 3H), 4.27 (m, 2H), 4.84 (m, 1H), 6.86 (m, 1H), 6.95 (m, 1H), 7.57 (m, 1H), 7.67 (m, 1H) 7.81 (m, 1H), 8.16 (m, 1H), 8.96 (s, 1H)

Example 6 N-[(1S) -1-carboxy-2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl ] Nicotinamide

N-[(1S) -1- (ethoxycarbonyl) -2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine obtained in Example 5 above The desired compound was prepared by hydrolysis from the 4-yl-carbonyl] nicotinamide by the method of Example 2.

Yield: 88%

m.p. : 210 to 212 ° C

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.51 (d, 6H), 3.79 (m, 4H), 3.93 (m, 1H), 4.14 (m, 2H), 4.38 (m, 2H), 4.52 (m, 1H), 4.64 (m, 1H), 5.35 (m, 1H), 7.65 (m, 1H), 7.84 (m, 2H), 8.65 (m, 1H), 9.48 (m, 1H), 9.82 (s, 1H)

Example 7 N-[(1R) -1- (ethoxycarbonyl) -2-hydroxyethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4 -Yl-carbonyl] nicotinamide hydrochloride

Using the D-serine ethyl ester hydrochloride in the same manner as in Example 1 to prepare the target compound as a crystal of the hydrochloride.

Yield: 65%

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.52 (m, 9H), 3.90 (m, 4H), 4.01 (m, 2H), 4.25 (m, 1H), 4.47 (m, 2H), 4.66 (m, 4H), 5.39 (m, 1H), 7.27 (m, 1H), 7.88 (m, 2H), 8.72 (m, 1H), 9.57 (m, 1H), 9.90 (m, 1H)

Example 8 N-[(1S, 2R) -2-hydroxy-1- (methoxycarbonyl) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine -4-yl-carbonyl] nicotinamide

L-threonine methyl ester hydrochloride was carried out in the same manner as in Example 3 always to prepare a target compound.

Yield: 73%

m.p. : 83 to 85 ° C

¹ H-NMR (CDCl ₃ ), ppm: δ1.22 (d, 6H), 1.31 (d, 3H), 3.05 (m, 2H), 3.14 (m, 2H), 3.51 (m, 3H), 3.77 ( s, 3H), 3.91 (m, 2H), 4.10 (m, 1H), 4.43 (m, 1H), 4.76 (m, 1H), 6.80 (m, 1H), 6.91 (m, 1H), 7.59 (m) , 2H), 7.73 (m, 1H), 8.12 (m, 1H), 8.93 (s, 1H)

Example 9 N-[(1S, 2R) -1-carboxy-2-hydroxypropyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl- Carbonyl] nicotinamide

N-[(1S, 2R) -2-hydroxy-1- (methoxycarbonyl) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] obtained in Example 8 above The target compound was prepared by hydrolysis from piperazin-4-yl-carbonyl] nicotinamide by the method of Example 2.

Yield: 93%

m.p. : 222-226 ℃

¹ H-NMR (DMSO-d ₆ ), ppm: δ1.07 (m, 9H), 3.31 (m, 4H), 3.54 (m, 3H), 3.84 (m, 2H), 4.25 (m, 1H), 4.49 (m, 1H), 7.19 (m, 2H), 7.37 (m, 1H), 7.74 (m, 1H), 8.53 (m, 1H), 9.11 (m, 1H)

Example 10 N-[(1S) -1- (ethoxycarbonyl) -3- (methylthio) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine -4-yl-carbonyl] nicotinamide hydrochloride

L-methionine ethyl ester hydrochloride was carried out in the same manner as in Example 1 to prepare the target compound as crystals of hydrochloride.

Yield: 62%

¹ H-NMR (CDCl ₃ ), ppm: δ 1.25 (m, 9H), 2.07 (s, 3H), 2.20 (m, 2H), 2.58 (m, 2H), 3.51 (m, 5H), 3.64 ( m, 2H), 3.96 (m, 2H), 4.20 (m, 2H), 4.83 (m, 1H), 7.19 (m, 2H), 7.59 (m, 1H), 7.71 (m, 1H), 7.80 (m , 1H), 8.21 (m, 1H), 8.96 (s, 1H)

Example 11 N-[(1S) -1-carboxy-3- (methylthio) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl- Carbonyl] nicotinamide

N-[(1S) -1- (ethoxycarbonyl) -3- (methylthio) propyl] -6- [1- [3- (isopropylamino) -2-pyridyl] obtained in Example 10 above The target compound was prepared from the piperazin-4-yl-carbonyl] nicotinamide hydrochloride by the method of Example 2 above.

Yield: 85%

m.p. : 172-175 ℃

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.47 (d, 6H), 2.33 (s, 3H), 2.49 (m, 1H), 2.64 (m, 1H), 2.94 (m, 2H), 3.70 (m, 4H), 3.88 (m, 1H), 4.11 (m, 2H), 4.34 (m, 2H), 5.30 (m, 1H), 7.59 (m, 1H), 7.77 (m, 2H), 8.60 (m, 1H), 9.41 (m, 1H), 9.77 (s, 1H)

Example 12 N-[(1S) -1- (ethoxycarbonyl) -2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl ] Piperazin-4-yl-carbonyl] nicotinamide

L-tyrosine ethyl ester hydrochloride was carried out in the same manner as in Example 3 to prepare a target compound.

Yield: 74%

m.p. : 93-95 ℃

¹ H-NMR (CDCl ₃ ), ppm: δ1.23 (m, 9H), 3.16 (m, 6H), 3.53 (m, 1H), 3.68 (m, 2H), 3.91 (m, 2H), 4.24 ( m, 3H), 4.98 (m, 1H), 6.78 (m, 3H), 6.84 (m, 4H), 7.63 (m, 1H), 7.71 (m, 1H), 8.08 (m, 1H), 8.81 (s , 1H)

Example 13: N-[(1S) -1-carboxy-2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine-4 -Yl-carbonyl] nicotinamide

N-[(1S) -1- (ethoxycarbonyl) -2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2- obtained in Example 12 above The desired compound was prepared by hydrolysis from pyridyl] piperazin-4-yl-carbonyl] nicotinamide by the method of Example 2.

Yield: 91%

m.p. : 126-130 degreeC

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.45 (d, 6H), 3.36 (m, 2H), 3.61 (m, 4H), 3.86 (m, 1H), 4.09 (m, 2H), 4.33 (m, 2H), 5.39 (m, 1H), 7.02 (m, 2H), 7.32 (m, 2H), 7.60 (m, 1H), 7.76 (m, 2H), 8.53 (m, 1H), 9.20 (m, 1 H), 9.58 (s, 1 H)

Example 14 N- (carbamoylmethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

Using the glycineamide hydrochloride was carried out in the same manner as in Example 3 to prepare a target compound.

Yield: 82%

m.p. : 169 ~ 170 ℃

¹ H-NMR (CDCl ₃ ), ppm: δ 1.24 (d, 6H), 3.17 (m, 4H), 3.56 (m, 1H), 3.66 (m, 2H), 3.94 (m, 2H), 4.11 ( m, 2H), 6.10 (m, 1H), 6.71 (m, 1H), 6.88 (m, 1H), 6.99 (m, 1H), 7.65 (m, 2H), 8.05 (m, 1H), 8.24 (m , 1H), 9.02 (s, 1H)

Example 15 N-[(1S) -1-carbamoyl-2-methylpropyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl ] Nicotinamide

L-valinamide hydrochloride was carried out in the same manner as in Example 3 to prepare a target compound.

Yield: 74%

m.p. : 115 ~ 120 ℃

¹ H-NMR (CDCl ₃ ), ppm: δ 1.05 (d, 6H), 1.24 (d, 6H), 2.24 (m, 1H), 3.18 (m, 4H), 3.55 (m, 1H), 3.66 ( m, 2H), 3.96 (m, 2H), 4.15 (m, 1H), 4.55 (m, 1H), 5.79 (m, 1H), 6.38 (m, 1H), 6.87 (m, 1H), 6.95 (m) , 1H), 7.39 (m, 1H), 7.68 (m, 2H), 8.20 (m, 1H), 8.96 (s, 1H)

Example 16: N-[(1S) -1-carbamoyl-2- (4-hydroxyphenyl) ethyl] -6- [1- [3- (isopropylamino) -2-pyridyl] piperazine- 4-yl-carbonyl] nicotinamide

L-tyrosineamide hydrochloride was carried out in the same manner as in Example 3 to prepare a target compound.

Yield: 67%

m.p. 135 to 139 ° C

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.45 (d, 6H), 3.35 (m, 1H), 3.46 (m, 1H), 3.70 (m, 4H), 3.88 (m, 1H), 4.07 (m, 2H), 4.41 (m, 2H), 5.35 (m, 1H), 7.04 (m, 2H), 7.32 (m, 2H), 7.59 (m, 1H), 7.75 (m, 2H), 8.53 (m, 1H), 9.26 (m, 1H), 9.62 (s, 1H)

Example 17 N- (ethoxycarbonylmethyl) -N-methyl-6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide Hydrochloride

N-methylglycine ethyl ester hydrochloride was carried out in the same manner as in Example 1 to prepare the target compound as crystals of hydrochloride.

Yield: 64%

¹ H-NMR (CDCl ₃ ), ppm: δ1.32 (m, 9H), 3.12 (m, 3H), 3.48 (m, 4H), 3.54 (m, 1H), 4.04 (m, 4H), 4.27 ( m, 4H), 7.31 (m, 2H), 7.83 (m, 2H), 7.96 (m, 1H), 8.65 (m, 1H)

Example 18 N- (carboxymethyl) -N-methyl-6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

N- (ethoxycarbonylmethyl) -N-methyl-6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotine obtained in Example 17 above From the amide hydrochloride, it was hydrolyzed by the method of Example 2 to prepare a target compound.

Yield: 92%

m.p. : 143 ~ 145 ℃

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.47 (d, 6H), 3.43 (s, 3H), 3.75 (m, 4H), 3.89 (m, 1H), 4.10 (m, 2H), 4.36 (m, 2H), 4.71 (s, 2H), 7.60 (m, 1H), 7.76 (m, 2H), 8.55 (m, 1H), 9.02 (m, 1H), 9.40 (m, 1H)

Example 19 N- (ethoxycarbonylethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide hydrochloride

Using the beta-alanine ethyl ester hydrochloride in the same manner as in Example 1 to prepare the target compound as a crystal of the hydrochloride.

Yield: 73%

¹ H-NMR (CDCl ₃ ), ppm: δ 1.29 (m, 9H), 2.71 (m, 2H), 3.53 (m, 4H), 3.67 (m, 1H), 3.74 (m, 2H), 3.89 ( m, 2H), 4.03 (m, 2H), 4.35 (m, 2H), 7.29 (m, 2H), 7.60 (m, 1H), 7.79 (m, 1H), 7.85 (m, 1H), 8.27 (m , 1H), 9.00 (s, 1H)

Example 20 N- (carboxyethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide

From N- (ethoxycarbonylethyl) -6- [1- [3- (isopropylamino) -2-pyridyl] piperazin-4-yl-carbonyl] nicotinamide hydrochloride obtained in Example 19 above Hydrolysis was carried out by the method of Example 2 to prepare a target compound.

Yield: 87%

m.p. : 200 to 203 ° C

¹ H-NMR (TFA-d ₁ ), ppm: δ 1.44 (d, 6H), 3.07 (m, 2H), 3.72 (m, 4H), 3.90 (m, 1H), 4.09 (m, 4H), 4.36 (m, 2H), 7.60 (m, 1H), 7.78 (m, 2H), 8.58 (m, 1H), 9.36 (m, 1H), 9.72 (s, 1H)

Experimental Example 1 In vitro Inhibitory Activity Test of Hepatitis B Virus Polymerase

Recently, the present inventors have filed a patent application by expressing and separating the recombinant polymerase protein of hepatitis B virus in E. coli and measuring the enzyme activity thereof (Korean Patent Application Nos. 94-3918 and 96-33998). The inventors have established a method for measuring reverse transcriptase activity of hepatitis B virus polymerase in vivo using this enzyme. The basic principle is the same as the enzyme-immunological method (ELISA), and the reaction is performed by incorporating biotin- and DIG- modified nucleotides into the substrate and then reacting the polymerized substrate with an anti-DIG antibody attached to peroxidase. Recognition method was used. 20 μl of hepatitis B virus polymerase, 20 μl of reaction mixture and 20 μl of sample were mixed and reacted at 14 ° C. to 30 ° C. for 18 to 24 hours, and compared with the result of control experiment without sample. Reverse transcriptase inhibitory activity was confirmed.

The inhibitory activity results of each sample are shown in Table 1 below.

Inhibitory Effect of Hepatitis B Virus (HBV) Polymerase on Reverse Transcriptase (HBV-RT) Activity compound R ₁ R ₂ Granularity R ₃ R ₄ n HBV-RT activity inhibitory effect (%) 1 µg / ml 0.1 µg / ml 0.01 µg / ml Example 1 CH ₃ CH ₂ O- H - H (CH ₃ ) ₂ CH- 0 77 64 30 Example 2 HO- H - H " 0 68 54 28 Example 3 CH ₃ CH ₂ O- CH _3- 1S H " 0 80 65 53 Example 4 HO- CH _3- 1S H " 0 79 57 22 Example 5 CH ₃ CH ₂ O- HOCH _2- 1S H " 0 83 68 54 Example 6 HO- HOCH _2- 1S H " 0 81 60 55 Example 7 CH ₃ CH ₂ O- HOCH _2- 1R H " 0 84 71 63 Example 8 CH ₃ O- 1S, 2R H " 0 85 70 60 Example 9 HO- 1S, 2R H " 0 83 61 45 Example 10 CH ₃ CH ₂ O- CH ₃ SCH ₂ CH _2- 1S H " 0 75 60 35

Table 1

(continue)

compound R ₁ R ₂ Granularity R ₃ R ₄ n HBV-RT activity inhibitory effect (%) 1 µg / ml 0.1 µg / ml 0.01 µg / ml Example 11 HO- CH ₃ SCH ₂ CH _2- 1S H (CH ₃ ) ₂ CH- 0 70 52 28 Example 12 CH ₃ CH ₂ O- (4-OH) C ₆ H ₄ CH _2- 1S H " 0 80 67 40 Example 13 HO- (4-OH) C ₆ H ₄ CH _2- 1S H " 0 79 65 54 Example 14 H ₂ N- H - H " 0 80 49 40 Example 15 H ₂ N- (CH ₃ ) ₂ CH- 1S H " 0 74 63 48 Example 16 H ₂ N- (4-OH) C ₆ H ₄ CH _2- 1S H " 0 55 47 20 Example 17 CH ₃ CH ₂ O- H - CH _3- " 0 64 51 27 Example 18 HO- H - CH _3- " 0 78 60 50 Example 19 CH ₃ CH ₂ O- H - H " One 80 54 43 Example 20 HO- H - H " One 83 65 31

Experimental Example 2 In vitro Inhibitory Activity Test of Human Immunodeficiency Virus (HIV) Reverse Transcriptase (HIV-RT)

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

First, 20 μl (40 ng) of HIV-RT were placed in a well coated with streptavidin, template / primer hybrid poly (A) oligo (dT) ₁₅ and DIG- (digoxygenin). 20 μl of a reaction mixture containing -dUTP, biotin-dUTP, and TTP was added, and then 20 μl of the sample to measure activity was added and reacted at 37 ° C. for 1 hour. The activity was compared as a control without the sample. At this time, the DNA is produced by the action of HIV-RT and contains digoxigenin and biotin-labeled nucleotides, thus binding to streptavidin coated on the bottom of the well. . After the reaction was completed, 250 μl of washing buffer solution (pH 7.0) was added to each well to remove remaining impurities, and washed five times for 30 seconds. 200 μl of anti-DIG-POD antibody was added thereto for 1 hour at 37 ° C. After reacting for a while, washed again with a washing buffer to remove impurities, and then reacted at room temperature for 30 minutes by adding 200 μl of ABTS ™, a substrate of POD (peroxidase), and then using an ELISA reader. Absorbance was measured at 405 nm to quantify reverse transcriptase activity and inhibitory activity.

Inhibitory activity of each sample is shown in Table 2 below.

Inhibitory Effect of Human Immunodeficiency Virus (HIV) on Reverse Transcriptase (HIV-RT) Activity compound R ₁ R ₂ Granularity R ₃ R ₄ n HIV-RT inhibitory effect (%) 1 µg / ml 0.1 µg / ml Example 1 CH ₃ CH ₂ O- H - H (CH ₃ ) ₂ CH- 0 72 49 Example 2 HO- H - H " 0 61 40 Example 4 HO- CH _3- 1S H " 0 65 47 Example 5 CH ₃ CH ₂ O- HOCH _2- 1S H " 0 73 38 Example 6 HO- HOCH _2- 1S H " 0 80 56 Example 7 CH ₃ CH ₂ O- HOCH _2- 1R H " 0 72 29 Example 9 HO- 1S, 2R H " 0 63 45 Example 11 HO- CH ₃ SCH ₂ CH _2- 1S H " 0 76 48 Example 15 H ₂ N- (CH ₃ ) ₂ CH- 1S H " 0 80 59 Example 18 HO- H - CH _3- " 0 59 30 Example 20 HO- H - H " One 70 41

Experimental Example 3 Cytotoxicity Test

Cytotoxicity test of the 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 according to the present invention and a pharmaceutically acceptable salt thereof was performed by an in vitro method using HepG2 cells. As a result, the 2,5-pyridinedicarboxylic acid derivative represented by the formula (1) according to the present invention and its pharmaceutically acceptable salts were all found to have excellent safety as CC ₅₀ ≥ 200 µM.

The 2,5-pyridinedicarboxylic acid derivative represented by Chemical Formula 1 and a pharmaceutically acceptable salt thereof according to the present invention have excellent anti-proliferative effects on HBV and HIV as well as no cytotoxicity, and thus hepatitis B and AIDS (acquired) Useful for the treatment and prevention of immunodeficiency syndrome).

Claims (5)

2,5-pyridinedicarboxylic acid derivative represented by the following formula (1) and pharmaceutically acceptable salts thereof. Formula 1 In Chemical Formula 1; R ₁ represents a linear or pulverized alkoxy group, a hydroxy group or an amino group having 1 to 4 carbon atoms, and R ₂ represents a hydrogen atom, a linear or pulverized alkyl group having 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Linear or pulverized hydroxyalkyl group, alkylthioalkyl group having 2 to 4 carbon atoms, phenyl group, benzyl group or hydroxybenzyl group, and when R ₂ represents a substituent other than a hydrogen atom, (R)-or (S) The stereospecificity of the form, R ₃ represents a hydrogen atom or a methyl group, R ₄ represents a hydrogen atom or a linear or pulverized alkyl group having 1 to 4 carbon atoms, n is 0 or 1. The method of claim 1, wherein in Chemical Formula 1, R ₁ is a methoxy group, an ethoxy group, a hydroxy group or an amino group; R ₂ is a hydrogen atom, a methyl group, isopropyl group, isobutyl group, hydroxymethyl group, 1-hydroxyethyl group, (2-methylthio) ethyl group, phenyl group, benzyl group, 4-hydroxybenzyl group, and R ₂ is Substituents other than hydrogen atoms have stereospecificity in the form of (R)-or (S)-; R ₃ is a hydrogen atom or a methyl group, R ₄ is a hydrogen atom, an ethyl group, isopropyl group or isobutyl group, n is 0 or 1, 2,5-pyridinedicarboxylic acid derivative. The pharmaceutical composition for the treatment and prevention of hepatitis B, characterized in that it contains a 2,5-pyridinedicarboxylic acid derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient. Formula 1 In Formula 1, R ₁ , R ₂ , R ₃ , R ₄ and n are as defined in claim 1, respectively. A pharmaceutical composition for the treatment and prevention of acquired immunodeficiency syndrome (AIDS), comprising 2,5-pyridinedicarboxylic acid derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. Formula 1 In Formula 1, R ₁ , R ₂ , R ₃ , R ₄ and n are as defined in claim 1, respectively. A method for producing a 2,5-pyridinedicarboxylic acid derivative represented by the following formula (1), which is prepared by reacting a nicotinic acid derivative represented by the following formula (2) with an amine compound represented by the following formula (3): Formula 1 In the above formulas, R ₁ represents a linear or pulverized alkoxy group or amino group having 1 to 4 carbon atoms, and R ₂ , R ₃ , R ₄ and n are as defined in claim 1, respectively.