KR19990085450A - New MEK Protein Inhibitors with Pyrazole Structure - Google Patents

Abstract

The present invention relates to a novel pyrazole derivative represented by the following formula (1), a preparation method thereof, and a MEK inhibitor composition containing the compound of formula (1) as an active ingredient:

Wherein R ₁ to R ₄ are as defined in the specification.

Description

New MEV Protein Inhibitors with Pyrazole Structure

The present invention relates to novel pyrazole derivatives represented by the following general formula (1), pharmaceutically acceptable salts or isomers thereof:

Formula 1

In the above formula

R ₁ represents a hydrogen atom, lower alkyl, benzyl, aryl, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom,

R ₂ is aryl monosubstituted or polysubstituted or unsubstituted by one or more substituents selected from halogen, lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing one or two nitrogen or sulfur atoms as hetero atoms. Indicates,

R ₃ represents hydrogen or aryl mono- or polysubstituted or unsubstituted by one or more substituents selected from lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom,

R ₄ represents lower alkoxycarbonyl, benzyl carbamoyl or amino monosubstituted or polysubstituted or unsubstituted by hydroxy, or monosubstituted or polysubstituted by one or more substituents selected from halogen, lower alkoxy and hydroxy Or aryl which is unsubstituted.

The present invention also relates to a method for preparing the compound of Formula 1 and a MEK (Mitogen Activated Protein Kinase Kinase) inhibitor composition comprising the compound as an active ingredient.

Research on cell signaling systems that control the growth and differentiation of cells, which are not only related to various diseases of the human body but also essential for the maintenance of life phenomena, is one of the most recent research fields. Dual MAPK (Mitogen-Activated Protein Kinase) signaling pathway is a signaling system that works when cells are stimulated by various mitogens such as growth factors, cytokines, or hormones. Is a signal transduction system that is well conserved in various organisms, from yeast to mammals (see Davis, RJ et. Al., TIBS, 19, 470, 1994). ).

Mammalian cells have various forms of MAPK pathways (Neiman, A. TIGS, 9, 390, 1993): Marshall, CJ Cell, 80, 179, 1995), in particular Ras, an important signaling agent The "Ras-MAPK" signaling system, which acts as one of the MAPK signaling systems, is currently well known. Many of the protein components involved in signal transduction are products of the protooncogene that we are well aware of, and are caused by mutations in these genes, namely, abnormal binding between proteins, overproduction of proteins by gene amplification, and changes in the activity of component proteins. All of the backs are known to cause cancer.

MEK protein is a protein that phosphorylates ERK-1 and ERK-2 proteins at the ends of the MAPK signaling system in the cytoplasm. It acts as an important mediator to transfer extracellular signals into the nucleus and differentiates the growth of cells. It modulates MAPK activity changes that induce a variety of physiological changes. The MEK protein activates the protein by phosphorylating threonine, 183th amino acid, and tyrosine, 185th amino acid, in the signal transduction process, and the activated ERK protein is threonine of myelin basic protein. Phosphorylation of residues (Marshall, CJ et. Al., Cell, 80, 179, 1995). Phosphorylation of threonine and tyrosine residues in ERK by the MEK protein plays a critical role in ERK activation and has been observed to be inactivated when these amino acid residues are substituted with other amino acid residues (Robins, DJ et. Al., J. Biol. Chem., 268, 5097, 1993).

Another type of MAPK, JNK (Jun kinase), was first discovered as a protein that phosphorylates the transcription factor c-Jun when cells are exposed to UV light (see Hibi, M. et. Al., Genes Dev., 7). , 2135, 1993). The amino acid sequencing of JNK revealed that JNK was not only very similar to ERKs, but also activated by phosphorylation of threonine and tyrosine residues. However, phosphorylation sites in JNK are known to be different from ERK as threonine-proline-tyrosine sites. JNK is also activated by tumor necrosis factor (TNF), interleukin-1 (IL-1), etc. in addition to UV (see Hibi, M. et. Al., Genes Dev., 7, 2135, 1993).

On the other hand, p38 is the most recently known phosphorylated protein of the MAPK family, and it was first discovered that tyrosine residues are phosphorylated when exposed to Gram-negative endotoxin (Han, J. et. Al., Science, 265, 808, 1994). Like JNK, p38 is also activated by TNF, IL-1, lipopolysaccharide (LPS), or osmotic stress. Although the substrate specificity for the p38 protein and the relationship between the ERK and JNK signaling pathways are not well known, p38 is most similar to the high-osmolarity glycerol response 1 (HOG1) in yeast, and the Thr-Gly-Tyr double It has a dual phosphorylation motif and is known to replace HOG1 function when expressed in yeast (Han, J. et. Al., Science, 265, 808, 1994). MEK activating ERK activates MAPKs such as JNK or p38 (Derijard, B. et. Al., Science, 267, 682, 1995), and 12-o-tetradecanoylpobol-13-acetate (12- o-tetradecanoylphorbol-13-acetate (TPA) strongly activates ERKs but does not activate JNK and p38, suggesting high specificity between them. On the other hand, actinomycin, okadaic acid and UV irradiation strongly activated JNK but did not significantly affect ERK, and the results showed that MEK protein was very specific for MAPK. Is showing. The relationship between JNK and p38 is not well known, but is known to use the same mitogens.

Abnormalities of proto-oncogenes such as ras, src, raf and mos are known to cause cancer by sustaining the active state of signaling agents in the MAPK signaling system (Gu, Z. et. Al., J. Virol., 69, 8051, 1995; Pelech, SL et. Al., TIBS, 17, 233, 1992). Therefore, knowing the correlation between cancer caused by mutations in the primary cancer genes such as ras and MAPK activity, it can be used to develop a treatment method for cancer by regulating MAPK activity, and develop a therapeutic agent that modulates the activity of MAPK. Therefore, it can be used as an effective anticancer agent for various types of cancers caused by defects in the uptream components of various signaling systems such as receptors, ras, and raf. However, despite this great importance, the development of a substance that can regulate the activity of MAPK is currently much less researched than the development of new drugs targeting other stages, and it is rarely in the practical use stage.

In order to develop a therapeutic agent for controlling the activity of MEK-1, the present inventors first succeeded in mass-purifying the high-purity expression of MEK-1 gene as an active form in insect cells and has already completed the patent application ( Reference: Korean Patent Application No. 97-10499). These results enabled the third-order structural analysis of MEK-1 protein by NMR and X-ray crystallography, and the development of new compounds with anticancer activity as MEK protein inhibitors. As a result, the present inventors have succeeded in developing a pyrazole derivative of Formula 1 having inhibitory activity against MEK-1 protein as a novel compound, and completed the present invention by confirming their anticancer activity.

Accordingly, an object of the present invention is to provide a novel pyrazole derivative of Formula 1, a pharmaceutically acceptable salt or isomer thereof having anticancer activity due to its inhibitory activity against MEK.

It is another object of the present invention to provide a method for preparing a compound of formula (1) or a salt thereof.

It is another object of the present invention to provide a MEK inhibitor composition containing a compound of Formula 1, a pharmaceutically acceptable salt or isomer thereof as an active ingredient together with a pharmaceutically acceptable carrier.

The present invention relates to novel pyrazole derivatives of formula (1), pharmaceutically acceptable salts or isomers thereof:

Formula 1

In the above formula

R ₁ represents a hydrogen atom, lower alkyl, benzyl, aryl, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom,

R ₂ is aryl monosubstituted or polysubstituted or unsubstituted by one or more substituents selected from halogen, lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing one or two nitrogen or sulfur atoms as hetero atoms. Indicates,

R ₃ represents hydrogen or aryl mono- or polysubstituted or unsubstituted by one or more substituents selected from lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom,

R ₄ represents lower alkoxycarbonyl, benzyl carbamoyl or amino monosubstituted or polysubstituted or unsubstituted by hydroxy, or monosubstituted or polysubstituted by one or more substituents selected from halogen, lower alkoxy and hydroxy Or aryl which is unsubstituted.

The term "lower alkyl" in the above definition for the compound of formula 1 refers to a straight or branched chain saturated hydrocarbon radical having 1 to 4 carbon atoms, such as methyl, ethyl, isopropyl, t-butyl, and the term "aryl" or "heteroaryl" Is a mono- or dicyclic aromatic group, of which "heteroaryl" refers to containing one or two heteroatoms as reduction.

Pharmaceutically acceptable salts of compounds of formula 1 according to the invention include pharmaceutically acceptable acid addition salts, such as aspartate, gluconate, glutamate, hydrochloride, para-toluenesulfonate or citrate, sodium, potassium or Salts with alkali metals such as lithium and the like, and salts with other acids or bases that are known and used for pyrazole compounds. These can be prepared by conventional conversion processes.

The compounds of formula (1) may also exist as enantiomers, racemates, diastereomers or mixtures thereof, so all these isomers and mixtures thereof are also included within the scope of the invention.

Among the compounds of Formula 1, which exhibit anticancer effects, more preferable compounds include R ₁ representing hydrogen atom, lower alkyl, benzyl, phenyl, or pyridyl, and R ₂ is phenyl mono- or di-substituted or unsubstituted by hydroxy. Represents; R ₃ represents hydrogen or phenyl; R ₄ is lower alkoxycarbonyl, a compound which represents monosubstituted, polysubstituted or unsubstituted benzylcarbamoyl or amino by hydroxy, or monophenyl, polysubstituted or unsubstituted phenyl by halogen.

Representative compounds in the compounds of Formula 1 according to the present invention may be mentioned those listed in Tables 1a to 1e below.

Compound number rescue designation 1-1 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole 1-2 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2H-pyrazole 1-3 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole 1-4 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-methyl-2H-pyrazole

1-5 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole 1-6 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2- (pyridin-4-yl) -2H-pyrazole 1-7 5-amino-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole 1-8 5-amino-3- (3,4-dihydroxyphenyl) -2H-pyrazole 1-9 3- (3,4-dihydroxyphenyl) -2,4-diphenyl-5-methoxycarbonyl-2H-pyrazole

1-10 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole 1-11 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole 1-12 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole 1-13 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole

1-14 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole 1-15 5- (3,4-dihydroxybenzylcarbamoyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole 1-16 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole 1-17 5- (2,4-difluorophenyl) -3- (4-hydroxyphenyl) -2H-pyrazole

1-18 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole 1-19 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole 1-20 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2- (pyridin-2-yl) -2H-pyrazole

The compound of formula 1 according to the present invention is (a) ring-opening reaction of a compound of formula (2a) in the presence of an acid catalyst in a solvent to obtain a compound of formula (1a), or (b) reducing the compound of formula (1b) in a solvent A compound of formula 1c may be obtained, or (c) a compound of formula 2b is hydrolyzed in the presence of an acid catalyst in a solvent to obtain a compound of formula 1d. The production method is another object of the present invention:

In the above formula

R ₁ to R ₄ are as defined above,

R ₁ ′ is the same as R ₁ , except for hydrogen

R ₅ represents a hydroxy protecting group.

As a solvent usable in the method (a) according to the present invention, one or more selected from methylene chloride, water and chlorobenzene may be mentioned, and the acid catalyst may be selected from BBr ₃ , PCl ₅ , AlCl ₃ , and BCl ₃ . More than one species can be mentioned. In particular, when PCl ₅ or AlCl ₃ is used as the acid catalyst, it is preferable to use water or chlorobenzene as a solvent, respectively. The reaction is preferably carried out for 1 to 2 hours in the temperature range of 25 to 50 ℃.

In the method (b), one or more selected from methanol, ethanol, tetrahydrofuran and ethyl acetate may be used as the solvent, and the reducing agent may be Pd (OH) ₂ / C and / or Pd / C at a pressure of 40 to 50 psi. Hydrogen gas may be used and reacted under these conditions for 6 to 12 hours.

In the method (c), as the solvent, one or more selected from methylene chloride, carbon tetrachloride and chloroform may be used, and as the acid catalyst, the same ones mentioned for the method (a) may be used. The reaction is preferably carried out for 1 to 2 hours in the temperature range of 25 to 30 ℃.

On the other hand, the compounds of Formulas 2a and 2b used as starting materials in the above methods can be easily prepared by applying the methods shown in Schemes 1 and 2, in addition to the desired substituents by combining the methods described in Starting compounds with can be prepared easily.

In Schemes 1 and 2

R ₁ , R ₃ and R ₄ are as defined above,

T ₁ represents hydrogen,

T ₂ represents oxo, or

T ₁ and T ₂ together with the carbon atoms to which they are attached may form an oxirane,

R ₂ ′ is benzo [1,3] dioxol-5-yl or Where R ₅ is as defined above,

X represents a leaving group, preferably halogen.

Hereinafter, the present invention will be described in more detail based on the following Preparation Examples and Examples. However, these preparation examples and examples are only to aid the understanding of the present invention, and the scope of the present invention is not limited by them in any sense.

Preparation Example 1 Preparation of (2,4-Difluorophenyl)-[3- (benzo [1,3] dioxol-5-yl) oxiran-2-yl] -methanone

To 0.54 g of 3- (benzo [1,3] dioxol-5-yl) -1- (2,4-difluoro-phenyl) -prop-2-en-1-one 1 ml of acetone and ethanol 3 After adding ml, 0.4 ml of 30% by weight hydrogen peroxide solution was added and 0.3 ml of 4N NaOH aqueous solution was added. The reaction solution was heated to 50 ° C. and stirred for 3 hours. After the reaction was terminated the white solid obtained was filtered and dried to give the title compound in 85% yield.

¹ H NMR (CDCl ₃ ): δ 7.98 (m, 1H), 7.03 (m, 1H), 6.90 (m, 2H), 6.82 (m, 2H), 5.99 (s, 2H), 4.22 (d, 1H) , 3.98 (s, 1H)

MS (FAB): 290 (M ⁺ + H ⁺ )

Preparation Example 2 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2-phenyl-2H-pyrazole

4 ml of ethanol (0.23 g of (2,4-difluorophenyl)-[3- (benzo [1,3] dioxol-5-yl) oxirane-2-yl] -methanone obtained in Preparation Example 1 It dissolved in and 0.086 ml of phenylhydrazine was added. Three drops of acetic acid were added to the mixture, and the mixture was heated to reflux for 1 hour. After completion of the reaction, distillation under reduced pressure, ethyl acetate and water were added, the organic layer was separated, dried over anhydrous magnesium sulfate, and then silica gel column chromatography (eluent: n-hexane / ethyl acetate = 9/1, v / v) yielded the title compound in 79% yield.

¹ H NMR (CDCl ₃ ): δ 8.10 (m, 1H), 7.23-7.10 (m, 4H), 6.95-6.64 (m, 6H), 5.90 (d, 2H), 5.15 (d, 2H)

MS (FAB): 377 (M ⁺ + H ⁺ )

Preparation Example 3 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2H-pyrazole

0.072 g of (2,4-difluorophenyl)-(3-benzo [1,3] dioxol-5-yl) oxirane-methanone obtained in Preparation Example 1 was dissolved in 1 ml of ethanol, and 0.013 ml of hydrazine. And 1 drop of acetic acid were added, followed by stirring for 1 hour. After the reaction was completed, the mixture was distilled under reduced pressure to remove ethanol, and the residue was separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3, v / v) to obtain the title compound in 20% yield.

¹ H NMR (CDCl ₃ ): δ 7.78 (m, 1H), 7.27-7.22 (m, 3H), 6.99-6.82 (m, 4H), 6.01 (s, 2H)

MS (FAB): 301 (M ⁺ + H ⁺ )

Preparation Example 4 Preparation of 4- (benzo [1,3] dioxol-5-yl) -2,4-dioxo-butyric acid methyl ester

0.195 g of dimethyl oxalate was dissolved in 0.75 mL of diethyl ether, 0.36 mL of 28% NaOMe solution was added, followed by stirring for 2 minutes. 0.25 g of 3 ', 4'-(methylenedioxy) acetophenone was added to the solution, stirred for 1 hour, and the resulting yellow solid was filtered to obtain the title compound in 99% yield.

¹ H NMR (d ₆ -DMSO): δ 7.35 (m, 1 H), 7.34 (s, 1 H), 6.89 (m, 1 H), 6.21 (s, 1 H), 6.03 (s, 2H), 3.64 (s, 3H), 3.55 (s, 1 H), 3.49 (s, 1 H).

MS (FAB): 251 (M ⁺ + H ⁺ )

Preparation Example 5 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole

0.0925 g of 4- (benzo [1,3] dioxol-5-yl) -2,4-dioxo-butyric acid methyl ester obtained in Preparation Example 4 was dissolved in 4 ml of ethanol, and 0.094 ml of hydrazine was added thereto, followed by heating to reflux. Stir overnight while stirring. After the reaction was completed, the reaction solution was extracted with ethyl acetate and water, followed by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3, v / v) to give the title compound in 73% yield. Obtained.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 7.51 (s, 1H), 7.21 (d, 2H), 6.98 (s, 1H), 6.88 (d, 1H), 6.01 (s, 2H), 4.49 ( s, 3 H)

MS (FAB): 247 (M ⁺ + H ⁺ )

Preparation Example 6 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole

0.0925 g of 4- (benzo [1,3] dioxol-5-yl) -2,4-dioxo-butyric acid methyl ester obtained in Preparation Example 4 was dissolved in 5 ml of ethanol, and then 0.063 ml of acetyl chloride was added thereto for 10 minutes. Was stirred. 0.040 ml of phenylhydrazine was added to the solution, and the mixture was heated to reflux for 3 hours, and then the solvent was removed by distillation under reduced pressure. Water and ethyl acetate were added to the residue, followed by extraction. The residue was separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 9/1, v / v) to obtain the title compound in 31% yield.

¹ H NMR (CDCl ₃ ): δ7.93 (d, 1H), 7.58 (s, 1H), 7.32 (m, 4H), 7.12 (m, 2H), 6.74 (m, 1H), 5.95 (s, 2H ), 3.89 (s, 3H)

MS (FAB): 323 (M ⁺ + H ⁺ )

Preparation Example 7 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-methyl-2H-pyrazole

0.064 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 5 was dissolved in 3 ml of DMF, and 0.012 g of NaH was added, followed by 10 minutes. Stirred. 0.019 ml of methane iodide was added to the reaction solution, and the mixture was stirred for 30 minutes. After completion of the reaction, the mixture was distilled under reduced pressure to remove DMF. The residue was separated by adding water and ethyl acetate to the residue, followed by drying over anhydrous magnesium sulfate, and silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3). , v / v) to give the title compound in 65% yield.

¹ H NMR (CDCl ₃ ): δ 7.30 (m, 2H), 7.00 (s, 1H), 6.84 (d, 1H), 6.00 (s, 2H), 4.19 (s, 3H), 3.90 (s, 3H )

MS (FAB): 249 (M ⁺ + H ⁺ )

Preparation Example 8 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2- (pyridin-4-yl) -2H-pyrazole

0.105 g of 4- (benzo [1,3] dioxol-5-yl) -2,4-dioxo-butyric acid methyl ester obtained in Preparation Example 4 and 0.050 g of 4-hydrazinopyridine were the same as in Preparation Example 5. To give the title compound in 7% yield.

¹ H NMR (CDCl ₃ ): δ 7.29 (s, 5H), 7.01 (d, 2H), 6.87 (d, 1H), 7.75 (m, 1H), 6.01 (s, 2H), 4.03 (s, 3H)

MS (FAB): 324 (M ⁺ + H ⁺ )

Preparation Example 9 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5- (t-butoxycarbonylamino) -2-phenyl-2H-pyrazole

To 0.20 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 6, 2 ml of tetrahydrofuran and 0.5 ml of water To this was added 0.10 g of NaOH and stirred overnight. After the reaction, the mixture was adjusted to pH 1 using 6N HCl, extracted with ethyl acetate, dried, and distilled under reduced pressure. 1 ml of t-butanol was added to the residue, 0.082 ml of dimethylphosphonyl azide and 0.053 ml of triethylamine were added, followed by stirring overnight to obtain the title compound in 21% yield.

¹ H NMR (CDCl ₃ ): δ 7.36-7.21 (m, 6H), 6.77-6.67 (m, 3H), 5.97 (s, 2H), 1.51 (s, 9H)

MS (FAB): 392 (M ⁺ + H ⁺ )

Preparation Example 10 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5- (t-butoxycarbonylamino) -2H-pyrazole

From 0.2098 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 5, the procedure was carried out in the same manner as in Preparation Example 9, and the title compound was 12. Obtained in% yield.

¹ H NMR (CDCl ₃ ): δ 7.27 (s, 1H), 7.12 (d, 2H), 6.85 (m, 1H), 6.55 (s, 1H), 6.01 (s, 2H)

MS (FAB): 316 (M ⁺ + H ⁺ )

Preparation Example 11 Preparation of 3- (benzo [1,3] dioxol-5-yl) -4-bromo-5-methoxycarbonyl-2-phenyl-2H-pyrazole

To 0.1877 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 6, 0.7 ml of acetic acid was added and 99% bromine water. Add 0.03 mL and stir overnight. After completion of the reaction, the mixture was basified to pH 12 using saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, distilled under reduced pressure and recrystallized from methanol to obtain the title compound in 55% yield.

¹ H NMR (CDCl ₃ ): δ 7.34-7.26 (m, 5H), 6.79 (d, 1H), 6.73 (m, 2H), 5.98 (s, 2H), 3.98 (s, 3H)

MS (FAB): 401 (M ⁺ + H ⁺ )

Preparation Example 12 Preparation of 3- (benzo [1,3] dioxol-5-yl) -2,4-diphenyl-5-methoxycarbonyl-2H-pyrazole

0.0774 g of 3- (benzo [1,3] dioxol-5-yl) -4-bromo-5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 11 was added to 2 ml of toluene. After dissolving, 0.0067 g of tetrakistriphenylphosphinepalladium was added thereto, followed by stirring for 10 minutes. 0.035 g of phenylboronic acid was added to the reaction solution, and 0.19 ml of an aqueous 2M sodium carbonate solution was added thereto, followed by heating to reflux overnight. After completion of the reaction, the solvent was removed by distillation under reduced pressure and the residue was separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3, v / v) to obtain the title compound in 32% yield.

¹ H NMR (CDCl ₃ ): δ 7.51-7.22 (m, 10H), 6.65 (d, 1H), 6.49 (m, 2H), 5.91 (s, 2H), 3.85 (s, 3H)

MS (FAB): 399 (M ⁺ + H ⁺ )

Preparation Example 13 Preparation of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-2H-pyrazole

0.21 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 5 was dissolved in 30 ml of DMF, and 0.40 g of NaH was added thereto. Stir for the next 10 minutes. 1.18 ml of benzyl bromide was added to the reaction mixture, followed by stirring at room temperature for 1 hour. After completion of the reaction, DMF was removed by distillation under reduced pressure, and ethyl acetate and water were added to the residue, followed by extraction. The solvent was removed by distillation under reduced pressure and recrystallized from diethyl ether to obtain the title compound quantitatively.

¹ H NMR (CDCl ₃ ): δ 7.40-7.21 (m, 7H), 7.07 (s, 1H), 6.85 (d, 1H), 5.95 (s, 2H), 5.75 (s, 2H), 3.85 (s, 3H)

MS (FAB): 335 (M ⁺ + H ⁺ )

Preparation 14 Preparation of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-4-bromo-5-methoxycarbonyl-2H-pyrazole

From 1.22 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 13, the same procedure as in Preparation Example 11 was carried out. The title compound was obtained in 81% yield.

¹ H NMR (CDCl ₃ ): δ 7.31-7.20 (m, 7H), 7.11 (s, 1H), 5.98 (s, 2H), 5.78 (s, 2H), 3.82 (s, 3H)

MS (FAB): 415 (M ⁺ + H ⁺ )

Preparation 15: Preparation of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-4-phenyl-2H-pyrazole

From 0.2636 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-4-bromo-5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 14 in Preparation Example 12 In the same manner as in the title compound was obtained in 31% yield.

¹ H NMR (CDCl ₃ ): δ 7.40-7.14 (m, 12H), 6.82 (s, 1H), 5.98 (s, 2H), 5.68 (s, 2H), 3.70 (s, 3H)

MS (FAB): 410 (M ⁺ + H ⁺ )

Preparation Example 16 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole

0.1841 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-4-phenyl-2H-pyrazole obtained in Preparation Example 15 was dissolved in 3 ml of methanol. Then 0.02 g of pd (OH) ₂ / C was added and reacted under a pressure of 50 psi of hydrogen for 3 hours. After the reaction was completed, the mixture was filtered and distilled under reduced pressure to obtain the title compound in 56% yield.

¹ H NMR (CDCl ₃ ): δ 7.40-7.32 (m, 5H), 7.15 (d, 1H), 7.07 (s, 1H), 6.85 (s, 1H), 6.08 (s, 2H), 3.85 (s, 3H)

MS (FAB): 321 (M ⁺ + H ⁺ )

Preparation Example 17 Preparation of 5- (2,4-difluorophenyl) -3- (4-methoxyphenyl) -2H-pyrazole

The title compound was obtained in 85% yield from 0.075 g of (2,4-difluorophenyl)-[3- (4-methoxyphenyl) oxiran-2-yl] -methanone in the same manner as in Preparation Example 3. Obtained.

¹ H NMR (CDCl ₃ ): δ 7.83 (m, 2H), 7.68 (m, 1H), 6.98 (m, 2H), 6.80 (s, 1H), 6.63 (m, 2H), 3.98 (s, 3H)

MS (FAB): 287 (M ⁺ + H ⁺ )

Preparation Example 18 Preparation of 3- (benzo [1,3] dioxol-5-yl) -5-benzylcarbamoyl-2-phenyl-2H-pyrazole

0.1 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 6 was diluted with THF / MeOH / H ₂ O (3 / 2/1, v / v / v) dissolved in 6 ml of a mixed solvent, 0.026 g of LiOH was added and stirred for 1 hour. After distillation under reduced pressure, the resulting acid compound was extracted with 1N HCl aqueous solution and ethyl acetate and the solvent was removed. The residue was dissolved in 5 ml of DMF, and 0.126 ml of benzylamine, 0.084 g of HOBt, 0.065 g of EDC and 0.043 ml of triethylamine were added and stirred for 1 hour. The DMF was removed by distillation under reduced pressure, extracted with 1N HCl aqueous solution and ethyl acetate, the solvent was removed by distillation under reduced pressure, and separated by silica gel column chromatography (eluent: methylene chloride / methanol = 99/1, v / v). Was obtained in 95% yield.

¹ H NMR (CDCl ₃ ): δ 7.36-7.28 (m, 10H), 7.00 (s, 1H), 6.73 (m, 2H), 6.63 (s, 1H), 5.96 (s, 2H), 4.65 (m, 2H)

MS (FAB): 398 (M ⁺ + H ⁺ )

Preparation Example 19 Preparation of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-benzylcarbamoyl-2H-pyrazole

0.1 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 13 was diluted with THF / MeOH / H ₂ O (3 / 2/1, v / v / v) dissolved in 6 ml of a mixed solvent, 0.025 g of LiOH was added and stirred for 1 hour. After distillation under reduced pressure, the resulting acid compound was extracted with 1N HCl aqueous solution and ethyl acetate and the solvent was removed. The residue was dissolved in 5 ml of DMF, and then 0.12 ml of benzylamine, 0.082 g of HOBt, 0.063 g of EDC and 0.041 ml of triethylamine were added and stirred for 1 hour. The DMF was removed by distillation under reduced pressure, extracted with 1N HCl aqueous solution and ethyl acetate, the solvent was removed by distillation under reduced pressure, and separated by silica gel column chromatography (eluent: methylene chloride / methanol = 99/1, v / v). Was obtained in 92% yield.

¹ H NMR (CDCl ₃ ): δ 7.38-7.20 (m, 11 H), 6.80 (m, 1 H), 6.65 (s, 1 H), 6.25 (m, 1 H), 5.95 (s, 2H), 5.78 (s, 2H), 4.57 (m, 2H)

MS (FAB): 412 (M ⁺ + H ⁺ )

Preparation Example 20 Preparation of 2-acetyl-3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole

0.124 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 5 was dissolved in 5 ml of DMF, and then 0.1 ml of triethylamine and 0.042 mL of acetyl chloride was added and stirred for 1 hour. The DMF was removed by distillation under reduced pressure and the residue was extracted with ethyl acetate, and then separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3, v / v) to obtain the title compound in 76% yield. It was.

¹ H NMR (CDCl ₃ ): δ 7.36-7.30 (m, 2H), 6.90 (s, 2H), 6.02 (s, 2H), 3.94 (s, 3H), 2.73 (s, 3H)

MS (FAB): 277 (M ⁺ + H ⁺ )

Preparation Example 21 Preparation of 1- (2,4-difluorophenyl) -3- (4-methoxyphenyl) -prop-2-en-1-one

The title compound was obtained in 85% yield from 0.5 g of 2,4-difluoroacetophenone and 0.346 ml of 4-methoxybenzaldehyde in the same manner as in Preparation Example 4.

¹ H NMR (CDCl ₃ ): δ 7.79 (m, 1H), 7.64 (m, 1H), 7.57 (m, 2H), 7.35 (m, 1H), 6.90 (m, 2H), 6.55 (m, 1H) , 6.48 (m, 1 H), 3.87 (s, 3 H)

MS (FAB): 275 (M ⁺ + H ⁺ )

Preparation Example 22 Preparation of (2,4-difluorophenyl)-[3- (4-methoxyphenyl) -oxirane-2-yl] -methanone

From 0.1 g of 1- (2,4-difluorophenyl) -3- (4-methoxyphenyl) -prop-2-en-1-one as in Preparation Example 1, the title compound was 89%. Obtained in yield.

¹ H NMR (CDCl ₃ ): δ 7.91 (m, 1H), 7.30 (m, 2H), 6.92 (m, 2H), 6.60 (m, 1H), 6.39 (m, 1H), 3.86 (s, 3H)

MS (FAB): 291 (M ⁺ + H ⁺ )

Preparation Example 23 of 3- (benzo [1,3] dioxol-5-yl) -5- (benzo [1,3] dioxol-5-ylmethyl) carbamoyl-2-phenyl-2H-pyrazole Produce

From 0.09 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 6, the same procedure as in Preparation Example 18 was carried out. The title compound was obtained in 79% yield.

¹ H NMR (CDCl ₃ ): δ 7.36-7.28 (m, 7H), 7.00 (s, 1H), 6.73 (m, 2H), 6.63 (s, 1H), 6.01 (s, 2H), 5.98 (s, 2H), 5.96 (s, 2H), 4.65 (m, 2H)

MS (FAB): 442 (M ⁺ + H ⁺ )

Preparation Example 24 Preparation of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-benzylcarbamoyl-4-phenyl-2H-pyrazole

From Preparation Example 18 from 0.08 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-4-phenyl-2H-pyrazole obtained in Preparation Example 15. In the same manner, the title compound was obtained in 89% yield.

¹ H NMR (CDCl ₃ ): δ 7.47-6.80 (m, 18H), 6.00 (s, 2H), 5.70 (m, 2H), 4.47 (m, 2H)

MS (FAB): 487 (M ⁺ + H ⁺ )

Preparation 25: Preparation of 3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2- (pyridin-2-yl) -2H-pyrazole

1 ml of ethanol (0.083 g of (2,4-difluorophenyl)-[3- (benzo [1,3] dioxol-5-yl) oxirane-2-yl] -methanone obtained in Preparation Example 1 It was dissolved in and 0.034 g of 2-hydrazinopyridine and 1 drop of acetic acid were added and then refluxed overnight. After completion of the reaction, the solvent was removed by distillation under reduced pressure and the residue was separated by silica gel column chromatography (eluent: methylene chloride / methanol = 99/1, v / v) to obtain the title compound in 50% yield.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 8.10 (m, 1H), 7.62 (m, 2H), 7.31 (m, 2H), 7.03 (m, 1H), 6.90 (m, 2H), 6.82 ( m, 2H)

MS (FAB): 378 (M ⁺ + H ⁺ )

Example 1: Synthesis of 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole (1-1)

0.093 g of 3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2-phenyl-2H-pyrazole obtained in Preparation Example 2 was diluted with 2.5 methylene chloride. To mL, 0.37 mL of 1.0M BBr ₃ solution was added thereto and stirred for 1 hour. 1 ml of methanol was added to the reaction solution to terminate the reaction, and the residue was separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 7/3, v / v) to obtain the title compound in 51% yield.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 8.21 (m, 1H), 7.31-7.11 (m, 4H), 6.99-6.70 (m, 6H), 5.20 (d, 2H)

MS (FAB): 365 (M ⁺ + H ⁺ )

Example 2: Synthesis of 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2H-pyrazole (1-2)

0.5 ml of methylene chloride was added to 0.0135 g of 3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2H-pyrazole obtained in Preparation Example 3, followed by 1M. 0.0675 mL BBr ₃ solution was added and stirred for 1 h. After the reaction was completed, 1 ml of methanol was added, distilled under reduced pressure, and then separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1/1, v / v) to obtain the title compound in 25% yield.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 7.81 (m, 1H), 7.20 (s, 1H), 7.12 (d, 1H), 6.97-6.90 (m, 3H), 6.80 (s, 1H)

MS (FAB): 289 (M ⁺ + H ⁺ )

Example 3: Synthesis of 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole (1-3)

0.0045 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 5 was dissolved in 0.2 ml of methylene chloride, followed by 1.0 M BBr. 0.029 ml of ₃ solution was added and stirred for 1 hour. 1 ml of methanol was added to the reaction mixture to complete the reaction, and the solvent was removed. The residue was separated by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1/1, v / v) to obtain the title compound. Obtained in 35% yield.

¹ H NMR (CD ₃ OD): δ 7.14 (s, 1 H), 7.06 (d, 1 H), 6.93 (s, 1 H), 6.83 (d, 1 H), 4.84 (s, 1 H)

MS (FAB): 235 (M ⁺ + H ⁺ )

Example 4: Synthesis of 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-methyl-2H-pyrazole (1-4)

0.0069 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-methyl-2H-pyrazole obtained in Preparation Example 7 was prepared as in Example 1. The title compound was obtained in 66% yield.

¹ H NMR (CDCl ₃ ): δ 4.49 (s, 1H), 7.23 (s, 1H), 7.12 (d, 1H), 7.02 (s, 1H), 6.86 (d, 1H), 4.18 (s, 3H ), 3.92 (s, 3 H)

MS (FAB): 261 (M ⁺ + H ⁺ )

Example 5: Synthesis of 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole (1-5)

0.0402 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole obtained in Preparation Example 6 was prepared in the same manner as in Example 1 The title compound was obtained in 20% yield.

¹ H NMR (CDCl ₃ ): δ 7.31-7.26 (m, 7H), 6.94 (s, 1H), 6.77 (d, 1H), 6.70 (s, 1H), 6.61 (d, 1H), 3.95 (s, 3H)

MS (FAB): 311 (M ⁺ + H ⁺ )

Example 6: Synthesis of 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2- (pyridin-4-yl) -2H-pyrazole (1-6)

Example 1 from 0.0363 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2- (pyridin-4-yl) -2H-pyrazole obtained in Preparation Example 8 In the same manner as in the title compound was obtained in 13% yield.

¹ H NMR (CD ₃ OD): δ 8.67 (d, 2H), 7.52 (d, 2H), 7.02 (s, 1H), 6.87 (d, 1H), 6.76-6.70 (m, 2H), 4.01 (s , 3H)

MS (FAB): 312 (M ⁺ + H ⁺ )

Example 7: Synthesis of 5-amino-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole (1-7)

In Example 1 from 0.0594 g of 3- (benzo [1,3] dioxol-5-yl) -5- (t-butoxycarbonylamino) -phenyl-2H-pyrazole obtained in Preparation Example 9 The title compound was obtained quantitatively in the same manner as in the following.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 7.41 (s, 1 H), 7.37-7.22 (m, 5 H), 6.75-6.71 (m, 2 H), 6.55 (d, 1 H)

MS (FAB): 268 (M ⁺ + H ⁺ )

Example 8: Synthesis of 5-amino-3- (3,4-dihydroxyphenyl) -2H-pyrazole (1-8)

The same procedure as in Example 1 was carried out from 0.0296 g of 3- (benzo [1,3] dioxol-5-yl) -5- (t-butoxycarbonylamino) -2H-pyrazole obtained in Preparation Example 10. To yield the title compound quantitatively.

¹ H NMR (CD ₃ OD): δ 7.17 (d, 1H), 7.12 (d, 1H), 6.89 (d, 1H)

MS (FAB): 192 (M ⁺ + H ⁺ )

Example 9: Synthesis of 3- (3,4-dihydroxyphenyl) -2,4-diphenyl-5-methoxycarbonyl-2H-pyrazole (1-9)

The same as in Example 1 from 0.0087 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-2,4-diphenyl-2H-pyrazole obtained in Preparation Example 12. To give the title compound in 94% yield.

¹ H NMR (CDCl ₃ ): δ 7.32-7.21 (m, 10H), 6.65 (d, 1H), 6.45 (d, 2H), 3.82 (s, 3H)

MS (FAB): 389 (M ⁺ + H ⁺ )

Example 10 Synthesis of 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole (1-10)

0.836 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-2H-pyrazole obtained in Preparation Example 13 was prepared as in Example 1. The title compound was obtained quantitatively.

¹ H NMR (CDCl ₃ ): δ 7.41-7.20 (m, 7H), 7.08 (s, 1H), 6.89 (d, 1H), 5.78 (s, 2H), 3.85 (s, 3H)

MS (FAB): 325 (M ⁺ + H ⁺ )

Example 11: Synthesis of 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole (1-11)

From 0.080 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-methoxycarbonyl-4-phenyl-2H-pyrazole obtained in Preparation Example 15 and In the same manner, the title compound was obtained in 38% yield.

¹ H NMR (CDCl ₃ ): δ 7.33-7.07 (m, 12H), 6.81 (s, 1H), 5.72 (s, 2H), 3.72 (s, 3H)

MS (FAB): 402 (M ⁺ + H ⁺ )

Example 12 Synthesis of 3- (3,4-Dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole (1-12)

From 0.08 g of 3- (benzo [1,3] dioxol-5-yl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole obtained in Preparation Example 16, the same procedure as in Example 1 was carried out. The title compound was obtained quantitatively.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 7.38-7.23 (m, 5H), 7.16 (d, 1H), 7.07 (s, 1H), 6.91 (s, 1H), 3.86 (s, 3H)

MS (FAB): 311 (M ⁺ + H ⁺ )

Example 13: Synthesis of 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole (1-13)

From 0.05 g of 3- (benzo [1,3] dioxol-5-yl) -5-benzylcarbamoyl-2-phenyl-2H-pyrazole obtained in Preparation Example 18, the procedure of Example 1 was repeated. The compound was obtained in 75% yield.

¹ H NMR (CDCl ₃ ): δ 7.37 (m, 5H), 7.21 (m, 1H), 7.10 (m, 1H), 6.92 (m, 2H), 4.64 (s, 2H)

MS (FAB): 385 (M ⁺ + H ⁺ )

Example 14 Synthesis of 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole (1-14)

From 0.025 g of 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole obtained in Example 16, the same procedure as in Preparation Example 16 was carried out to give 60% of the title compound. Obtained in yield.

¹ H NMR (CDCl ₃ ): δ 7.37-7.22 (m, 10H), 6.97 (m, 2H), 6.71 (m, 1H), 6.41 (m, 1H), 4.64 (m, 2H)

MS (FAB): 310 (M ⁺ + H ⁺ )

Example 15 Synthesis of 5- (3,4-Dihydroxybenzylcarbamoyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole (1-15)

3- (benzo [1,3] dioxol-5-yl) -5- (benzo [1,3] dioxol-5-ylmethyl) carbamoyl-2-phenyl-2H-pyra obtained in Preparation Example 23 The title compound was obtained in 45% yield from 0.05 g of sol in the same manner as in Example 1.

¹ H NMR (CDCl ₃ + CD ₃ OD): δ 7.38-7.30 (m, 5H), 6.90-6.62 (m, 6H), 6.51 (m, 1H), 4.46 (s, 2H)

MS (FAB): 418 (M ⁺ + H ⁺ )

Example 16: Synthesis of 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole (1-16)

The same procedure as in Example 1 was carried out from 0.07 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-benzylcarbamoyl-2H-pyrazole obtained in Preparation Example 19. The compound was obtained in 75% yield.

¹ H NMR (CDCl ₃ ): δ 7.24-7.11 (m, 10H), 6.97 (m, 1H), 6.70 (m, 2H), 6.46 (s, 1H), 5.61 (s, 2H), 4.42 (m, 2H)

MS (FAB): 400 (M ⁺ + H ⁺ )

Example 17 Synthesis of 5- (2,4-difluorophenyl) -3- (4-hydroxyphenyl) -2H-pyrazole (1-17)

0.05% of the title compound by the same procedure as in Example 1 from 0.05 g of 5- (2,4-difluorophenyl) -3- (4-methoxyphenyl) -2H-pyrazole obtained in Preparation Example 17. Obtained in yield.

¹ H NMR (CDCl ₃ ): δ 7.83 (m, 2H), 7.68 (m, 1H), 6.98 (m, 2H), 6.80 (s, 1H), 6.63 (m, 2H)

MS (FAB): 273 (M ⁺ + H ⁺ )

Example 18 Synthesis of 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole (1-18)

The same as in Example 1 from 0.78 g of 3- (benzo [1,3] dioxol-5-yl) -2-benzyl-5-benzylcarbamoyl-4-phenyl-2H-pyrazole obtained in Preparation Example 24. To give the title compound in 81% yield.

¹ H NMR (CDCl ₃ ): δ 7.7.40-6.75 (m, 18H), 5.75 (m, 2H), 4.50 (m, 2H)

MS (FAB): 476 (M ⁺ + H ⁺ )

Example 19 Synthesis of 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole (1-19)

The same procedure as in Preparation Example 16 was carried out from 0.056 g of 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole obtained in Example 18. The compound was obtained in 50% yield.

¹ H NMR (CDCl ₃ ): δ 7.40-6.82 (m, 13 H), 4.55 (m, 2 H)

MS (FAB): 386 (M ⁺ + H ⁺ )

Example 20: 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2- (pyridin-2-yl) -2H-pyrazole (1-20) synthesis

3- (benzo [1,3] dioxol-5-yl) -5- (2,4-difluorophenyl) -2- (pyridin-2-yl) -2H-pyrazole obtained in Preparation Example 25 The title compound was obtained in 70% yield from 0.05 g as in Example 1.

¹ H NMR (CDCl ₃ ): δ 8.10 (m, 1H), 7.60 (m, 2H), 7.26 (m, 2H), 6.98 (m, 1H), 6.87 (m, 2H), 6.76 (m, 2H)

MS (FAB): 366 (M ⁺ + H ⁺ )

Experimental Example: MEK Protein Inhibitory Activity Assay

MEK protein inhibition activity assay was carried out according to the method described in Korean Patent Application No. 97-32264, the results are shown in Table 2 together with the molecular weight of the compound according to the invention.

Compound number Molecular Weight IC ₅₀ (μM) One 364 0.9 2 288 11.8 3 234 4.8 4 260 2.0 5 310 1.4 20 365 10.2

From the results of Table 2, it can be seen that the novel pyrazole derivatives according to the present invention possess excellent inhibitory effect on MEK protein activity, and thus can be usefully used as an anticancer agent.

Claims (5)

The present invention relates to novel pyrazole derivatives represented by the following general formula (1), pharmaceutically acceptable salts or isomers thereof: Formula 1 In the above formula R ₁ represents a hydrogen atom, lower alkyl, benzyl, aryl, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom, R ₂ is aryl monosubstituted or polysubstituted or unsubstituted by one or more substituents selected from halogen, lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing one or two nitrogen or sulfur atoms as hetero atoms. Indicates, R ₃ represents hydrogen or aryl mono- or polysubstituted or unsubstituted by one or more substituents selected from lower alkyl, lower alkoxy and hydroxy, or heteroaryl containing a nitrogen or sulfur atom as a hetero atom, R ₄ represents lower alkoxycarbonyl, benzyl carbamoyl or amino monosubstituted or polysubstituted or unsubstituted by hydroxy, or monosubstituted or polysubstituted by one or more substituents selected from halogen, lower alkoxy and hydroxy Or aryl which is unsubstituted. The compound of claim 1, wherein R ₁ represents a hydrogen atom, lower alkyl, benzyl, phenyl, or pyridyl, and R ₂ represents phenyl mono- or di-substituted or unsubstituted by hydroxy; R ₃ represents hydrogen or phenyl; R ₄ represents lower alkoxycarbonyl, benzylcarbamoyl or amino mono- or polysubstituted or unsubstituted by hydroxy, or mono- or polysubstituted or unsubstituted phenyl by halogen. The method of claim 1, 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole; 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2H-pyrazole; 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole; 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-methyl-2H-pyrazole; 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2-phenyl-2H-pyrazole; 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2- (pyridin-4-yl) -2H-pyrazole; 5-amino-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole; 5-amino-3- (3,4-dihydroxyphenyl) -2H-pyrazole; 3- (3,4-dihydroxyphenyl) -2,4-diphenyl-5-methoxycarbonyl-2H-pyrazole; 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-2H-pyrazole; 2-benzyl-3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole; 3- (3,4-dihydroxyphenyl) -5-methoxycarbonyl-4-phenyl-2H-pyrazole; 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole; 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole; 5- (3,4-dihydroxybenzylcarbamoyl) -3- (3,4-dihydroxyphenyl) -2-phenyl-2H-pyrazole; 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -2H-pyrazole; 5- (2,4-difluorophenyl) -3- (4-hydroxyphenyl) -2H-pyrazole; 2-benzyl-5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole; 5-benzylcarbamoyl-3- (3,4-dihydroxyphenyl) -4-phenyl-2H-pyrazole; or 5- (2,4-difluorophenyl) -3- (3,4-dihydroxyphenyl) -2- (pyridin-2-yl) -2H-pyrazole. (a) ring-opening the compound of formula 2a in the presence of an acid catalyst in a solvent to obtain a compound of formula 1a, or (b) reducing the compound of formula 1b in a solvent to obtain a compound of formula 1c; c) A process for preparing the compound of formula 1 as defined in claim 1, characterized in that the compound of formula 2b is hydrolyzed in the presence of an acid catalyst in a solvent to obtain a compound of formula 1d: Formula 2a Formula 1a Formula 1b Formula 1c Formula 2b Formula 1d In the above formula R ₁ to R ₄ are as defined in claim 1, R ₁ ′ is the same as R ₁ , except for hydrogen R ₅ represents a hydroxy protecting group. MEK inhibitor composition comprising a compound as defined in claim 1, a pharmaceutically acceptable salt or isomer thereof as an active ingredient together with a pharmaceutically acceptable carrier.