KR100850969B1 - 1-[1-3,4-dialkoxyphenyl-alkyl]-1H-pyrazole compounds, process for preparing and pharmaceutical composition comprising the same - Google Patents

Abstract

The present invention provides a novel 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof, a method for preparing the same, and For the treatment of inflammation-related diseases, including asthma and chronic obstructive pulmonary disease, characterized in that it contains as an active ingredient and for the prevention and treatment of degenerative brain diseases such as arthritis, atopic dermatitis, leukemia and various cancers and Alzheimer's disease Or it relates to a pharmaceutical composition for memory enhancement.

[Formula 1]

1- [1- (3,4-Dialkoxyphenyl) -alkyl] -1H-pyrazole compound, PDE-4, asthma, chronic obstructive pulmonary disease, inflammatory diseases, arthritis, atopic dermatitis, leukemia, cancer, central nervous system Disease, Alzheimer's.

Description

Novel 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1H-pyrazole compound, preparation method thereof and pharmaceutical composition comprising the same {1- [1- (3,4-dialkoxyphenyl)- alkyl] -1H-pyrazole compounds, process for preparing and pharmaceutical composition comprising the same}

The present invention relates to a 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound, a pharmaceutically acceptable salt thereof, a method for preparing the same, and an asthma and chronic obstructive pulmonary disease comprising the same. (Chronic Obstructive Pulmonary Disease) and the composition for treating inflammation-related diseases, including arthritis, atopic dermatitis, leukemia and various degenerative brain diseases such as cancer and Alzheimer's disease, or a pharmaceutical composition for improving memory.

The present invention relates to a 1- [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole compound, a pharmaceutically acceptable salt thereof, a method for preparing the same, and an asthma and chronic obstructive pulmonary disease comprising the same. (Chronic Obstructive Pulmonary Disease), arthritis, inflammation-related diseases including atopic dermatitis and brain diseases, such as Alzheimer's, depression, memory loss and pharmaceutical compositions of various cancer treatments, including leukemia.

PDE-4 inhibitors on which the present invention is based, various compounds have been studied for the development of treatment for asthma and chronic obstructive pulmonary disease [Peter Norman,Expert Opin . Ther . Patents . 2002,12 (1), pp 93-111], Rolipram [EP 0660711, 5 July 1995], Cilomilast [USP 6,013,827, 11 January 2000] and Roflumilast [USP 5,712,298 Jan. 27, 1998] And the like are known. Rolipram, the first of these compounds to enter the clinic, was stopped due to side effects such as weak clinical effects and vomiting. In addition, Cilomilast is a compound that has ceased development as an asthma treatment due to the low therapeutic effect on asthma and clinical trials for Chronic Obstructive Pulmonary Disease (COPD) [Peter Norman,Expert Opin . Ther . Patents 2002,12 (1), 93-111; Compton C., Edelson JD., Cedar E ,.Am . J. Respir . Crit . Care Med . 2001,163, A909]. Roflumilast [Zheng Huang, Yves Ducharme, Dwight Macdonald and Annette Robichaud, developed by Altana, Germany]Current Opinion in Chemical Biology 2001,5,432-438) has a high PDE-4 inhibitory effect (IC₅₀ = 0.8 nM) Although it is known that the effects on asthma and COPD are excellent in animal studies, [Armin Hatzelman and Christian Schudt,The Journal of Pharmacology and Experimentalal Therapeutics 2001,297 (1), 267-279; Daniela S. Bundschuh, Manfrid Eltze, Johannes Barsig, Lutz Wollin, Armin Hatzelmann, and Rolf Beume,The Journal of Pharmacology and Experimentalal Therapeutics 2001,297 (1, 280-290] Due to side effects such as composition and headache, asthma treatment was canceled in 2005 in EMEA.

PDE-4 inhibitors treat arthritis in addition to the inflammatory-related lung diseases such as asthma and chronic obstructive pulmonary disease (USP 2003/0092706 A1), atopic dermatitis, leukemia, various cancers (Miles D. Houslay, Peter Schafer and Kam) YJ Zhang, Drug Discovery Today , 2005 , 10 (22), 1503-1519) and depression treatment effects (USP 4,178,449, 11 December 1979), Alzheimer's and other brain diseases treatment effects (Sophie L. Rovner, C & EN , 38, 2005 2 May 21) is also known. Therefore, as illustrated above, the development of a new drug based on PDE-4 inhibition is a situation that requires the development of treatments for various applications and improvement of side effects such as vomiting and headache.

Therefore, the inventors of the present invention for the development of a new structure with excellent in vitro and in vivo effects for the development of inflammatory diseases, brain diseases and anti-cancer agents, including asthma and rheumatoid arthritis, atopic dermatitis and the like based on PDE-4 inhibition 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole derivatives were prepared, and the compounds according to the present invention showed excellent inhibitory activity against PDE-4 enzyme as a result of biochemical and pharmacological tests. And high selectivity, disease model animal experiments have been shown to have excellent asthma and rheumatoid arthritis treatment effect.

Thus, it is to provide a 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound and a pharmaceutically acceptable salt thereof, and as another object a novel 1- according to the present invention. Inflammation-related diseases and arthritis, including methods for preparing [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole compounds, and asthmatic and chronic obstructive pulmonary diseases, including the same It is to provide a pharmaceutical composition for the prevention and treatment of atopic dermatitis, leukemia and various degenerative brain diseases such as cancer and Alzheimer's disease or memory enhancement.

The present invention is a novel 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof, a method for preparing the same And pharmaceutical compositions for preventing and treating degenerative brain diseases such as asthma and chronic obstructive pulmonary disease, including arthritis, atopic dermatitis, leukemia and various cancers and Alzheimer's, or memory enhancement, including asthma and chronic obstructive pulmonary disease. It is about.

[Formula 1]

[In Formula 1 and R ₁ And R ₂ independently of one another is a hydrogen atom, straight or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl, halogen substituted linear or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group, aryl or heteroaryl, ar (C ₁ -C ₅ ) alkyl, ar (C ₁ -C ₅ ) alkoxy, or with R ₁ R ₂ may be connected to each other (C ₁ -C ₃ ) alkylene, and R ₁ and (C ₃ -C ₇ ) cycloalkyl of R ₂ , The cycloalkyl of the (C ₄ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group may be a heterocyclic ring substituted with a heteroatom selected from oxygen, nitrogen or sulfur, and R ₁ and The substituent of R ₂ may be further substituted with halogen, hydroxy; R _3, R ₄ and R ₅ independently of one another are hydrogen, phenyl, pyridinyl, N-oxypyridinyl, straight or branched saturated and unsaturated (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cyclo Alkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, halogen atom, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino, mono or di (C ₁ -C ₇ ) Alkylaminocarbonyl, (C ₃ -C ₇ ) cycloalkylamino, 3- to 7-membered saturated or unsaturated heterocycles containing in the heterocycle, guanidinyl, ureido, benzyl , Benzyloxy, formyl, (C ₁ -C ₇ ) alkanoyl, (C ₁ -C ₇ ) alkoxy, (C ₁ -C ₇ ) alkoxy (C ₁ -C ₇ ) alkyl, hydroxy (C ₁ -C ₇ ) Alkyl, carboxyl, (C ₁ -C ₇ ) alkoxycarbonyl, wherein (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, phenyl, pyridinyl, N-oxypyridinyl, ar (C ₁ -C ₅ ) alkyl and ar (C ₁ -C ₅ ) alkoxy (C ₁ -C ₇ ) alkyl group, (C ₃ -C ₇ ) cycloalkyl group, (C ₄ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group, (C ₁ -C ₇ ) alkoxy, 5-membered or 6 At least one member selected from a heterocyclic ring, halogen, nitro, cyano, hydroxy, carboxyl or amino group, mono or di (C ₁ -C ₇ ) alkylamino may be further substituted; R ₆ is straight or branched saturated and unsaturated (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, phenyl, phenoxy, benzyl, benzyloxy, (C ₁ -C ₇ ) alkoxy, (C ₁ -C ₇ ) alkoxy (C ₁ -C ₇ ) alkyl, alkyl, cycloalkyl of R ₆ , The phenyl group is a (C ₁ -C ₇ ) alkyl group, (C ₃ -C ₇ ) cycloalkyl group, (C ₄ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group, (C ₁ -C ₇ ) alkoxy, halogen, One or more selected from nitro, cyano, hydroxy, carboxyl or amino groups, mono or di (C ₁ -C ₇ ) alkylamino may be further substituted, provided that R _{3 ,} R ₄ And R ₅ are both hydrogen.

Among the novel 1- [1- (3,4-alkoxyphenyl) -alkyl] -1 H -pyrazole compounds represented by Formula 1 according to the present invention, R ₁ and R ₂ are independently hydrogen atom, a saturated or unsaturated straight or branched chain (C ₁ - C ₇₎ alkyl, substituted by halogen or straight-chain saturated or unsaturated grinding (C ₁ - C ₇₎ alkyl, (C ₃ - C ₇ ) cycloalkyl group, (C ₃ -C ₈ ) cycloalkylmethyl or allyl group, particularly preferably R ₁ and R ₂ are each independently methyl, difluoromethyl, cyclopentyl or cyclopropylmethyl , R ₃ is cyclopentyl, cyclohexyl, phenyl, pyridinyl, N-oxypyridinyl, morpholino, piperazinyl, furanyl, or thiophenyl and the cyclopentyl, cyclohexyl, phenyl and pyridinyl or N-oxypyridinyl is hydroxy, Halogen, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino, (C ₁ -C ₇ ) alkoxy (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkylamino , Morpholino, N-oxymorpholino, N-oxypiperazinyl, piperazinyl, furanyl, thiophenyl, guanidinyl, ureido, carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl , Can be substituted with carbamoyl; R ₄ And R ₅ is independently of each other a hydrogen atom, (C ₁ -C ₇ ) alkyl or phenyl, which may be further substituted with carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl, carbamoyl.

More specifically, R _{3 ,} R ₄ of the 1- [1- (3,4-alkoxyphenyl) -alkyl] -1 H -pyrazole compound according to the present invention And R ₅ can be exemplified by the following substituents.

3-pyridinyl,

N-oxypyridin-3-yl

3- (ethoxycarbonyl) phenyl

3-carboxyphenyl

4-pyridinyl

4- (acetamido) phenyl

N-oxypyridin-4-yl

3-aminophenyl

3- (benzyloxy) phenyl

3-hydroxyphenyl

3-((ethoxycarbonyl) methoxy) phenyl

3- (benzamido) phenyl

3- (methoxy) phenyl

3- (dibenzylamino) phenyl

3- (benzylamino) phenyl

3- (methylamino) phenyl

3- (2- (N, N-dimethylaminoethoxy) phenyl

3- (dimethylamino) phenyl

3-chlorophenyl

3-bromophenyl

3- (pyrrolidin-1-yl) phenyl

3- (piperidin-1-yl) phenyl

Phenyl

3- (methanesulfonylamino) phenyl

3-((N, N-di- (methanesulfonyl)) amino) phenyl

4- (benzyloxy) phenyl

3-cyanophenyl

3- (2H-tetrazol-5-yl) phenyl

4-hydroxyphenyl

4-((ethoxycarbonyl) methoxy) phenyl

4-methoxyphenyl

3- (i-propoxycarbonyl) phenyl

4- (carboxymethoxy) phenyl

3- (trifluoromethanesulfonylamino) methyl

3-((N, N-di- (trifluoromethanesulfonyl) amino) methyl

4- (ethoxycarbonyl) phenyl

4-carboxyphenyl

4- (2-chloropyridinyl) phenyl

4-cyanophenyl

4- ( 1H -tetrazol-5-yl) phenyl

3,5-dichloropyridin-4-yl

4- (N-oxypyridinyl)

4-nitrophenyl

3-amino-4-chlorophenyl

4-aminophenyl

3-cyano-4-fluorophenyl

3-bromo-4-methoxyphenyl

3,5-dibromophenyl

(3-carboxy-4-methoxy) phenyl

3-ethylaminophenyl

(3-amino-4-methyl) phenyl

(3-nitro-4-methyl) phenyl

(3-ethoxycarbonyl-4-methyl) phenyl

(3-carboxy-4-methyl) phenyl

3-ureidophenyl

3-((ethoxycarbonyl) methylamino) phenyl

3-((carboxy) methylamino) phenyl

3-morpholinophenyl

3- (tetrahydro-2,4,6-trioxopyrimidin-1 (2H) -yl) phenyl

3-((ethoxycarbonylmethyl) carbamoyl) phenyl

3- (1- (ethoxycarboxy) -2-phenylethylcarbamoyl) phenyl

3-carbamoylphenyl

(3-carboxy-4-fluoro) phenyl

Cyclohexyl

3-cyanocyclohexyl

3-ethoxycarbonylphenyl

3-carboxycyclohexyl

4-ethoxycarbonylphenyl

Morpholino

4- (4-toluenesulfonyl) piperazin-1-yl

3-((1-methoxycarbonyl-2-methyl) butylcarbamoyl) phenyl

3-((1-methoxycarbonyl) ethylcarbamoyl) phenyl

3-((1-methoxycarbonyl-2-methyl) propylcarbamoyl) phenyl

3-((1-carboxy-2-methyl) propylcarbamoyl) phenyl

3-((1-carboxy-3-methyl) butylcarbamoyl) phenyl

3-((1-carboxy) ethylcarbamoyl) phenyl

3-((2,2,2-trifluoroethoxy) carbonyl) phenyl

3-((1- (propyl-2-oxycarbonyl) -2-methyl) propylcarbamoyl) phenyl

3-((1-ethoxycarbonyl-3-methyl) butylcarbamoyl) phenyl

3-((1- (propyl-2-oxycarbonyl) -3-methyl) butylcarbamoyl) phenyl

3-((pivaloyloxy) methoxycarbonyl) phenyl

3-chloro- (N-oxypyridin-4-yl)

Furan-2-yl

Furan-3-yl

5-methylfuran-2-yl

3-chlorofuran-2-yl

5-chlorofuran-2-yl

Thiophen-2-yl

Thiophen-3-yl

3-methylthiophen-2-yl

5-methylthiophen-2-yl

3-chlorothiophen-2-yl

5-chlorothiophen-2-yl

5-bromothiophen-2-yl

2,5-dichlorothiophen-3-yl

R ₆ in the substituent of the compound of formula 1 according to the present invention is specifically (C ₁ -C ₇ ) alkyl group or phenyl, and the alkyl group is carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl, carbamoyl, Hydroxy, At least one selected from halogen, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino or (C ₁ -C ₇ ) alkoxy can be substituted, in particular R ₆ is methyl, Ethyl, n-propyl, i-propyl, n-butyl, ethoxycarbonylmethyl, carboxymethyl, hydroxymethyl, 2-hydroxyethyl, cyanomethyl or carbamoylmethyl.

The compound of formula 1 according to the present invention typically includes the following compounds.

[In the compound, R ₆ is selected from methyl, ethyl, ethoxycarbonylmethyl, carboxymethyl, cyanomethyl.]

The method for preparing a 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound according to the present invention includes a halobenzyl compound represented by Chemical Formula 2 as shown in Scheme 1 below. It can be exemplified by the reaction route of the pyrazole compound represented by the formula (3), and the following preparation method does not limit the method for preparing the compound of formula (1) according to the present invention, modifications of the following preparation method will be apparent to those skilled in the art. Unless otherwise stated, the definitions of substituents in the following schemes are the same as in Formula 1.

Scheme 1

As shown in Scheme 1, the habenzyl compound of Formula 2 may be prepared by the same method as in Scheme 2 below.

Scheme 2

The 3,4-dialkoxybenzyl halogen compound represented by the formula (2) is alkylated with the corresponding aldehyde to form benzyl alcohol compound 5, as shown in Scheme 2, and then halogenated compound 5 to obtain the benzylhalogen compound of starting material 2. I can make it.

In addition to the ketone compound 6 as a pyrazole compound represented by the formula (3) it is shown in Scheme 3 N, N - dimethylformamide dimethyl acetal (N, N -dimethylformamidedimethylacetal) by reacting the compound 7 enamine type (enamine -type), and pyrazole compounds such as 8 may be prepared by reacting hydrazine with compound 7, but the pyrazole compound may be prepared by various other methods.

Scheme 3

Meanwhile, the 1- [1- (3,4-alkoxyphenyl) -alkyl] -1 H -pyrazole compound represented by Chemical Formula 1 prepared according to the present invention was biochemically and pharmacologically tested to PDE-4 enzyme. In addition to the excellent inhibitory activity and high selectivity, the disease model showed excellent asthma and rheumatoid arthritis treatment effects in animal experiments, but the side effect test using ferret model animals showed no related side effects such as vomiting or diarrhea.

Compound represented by the formula (1) according to the present invention as a pharmaceutical composition for the treatment of inflammation-related diseases including bronchial asthma and chronic obstructive pulmonary disease and the treatment, prevention of diseases of the central nervous system including Alzheimer's Suitable salts of the compound of formula (I) which are suitable for use and suitable for use in the medicament may include those formed with both organic and inorganic acids or bases. Pharmaceutically acceptable acid addition salts include hydrochloric acid, bromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, trifluoroacetic acid, triphenylacetic acid, phenylacetic acid, substituted phenylacetic acid, for example methoxy Phenylacetic acid, sulfamic acid, sulfanilic acid, succinic acid, oxalic acid, fumaric acid, maleic acid, malic acid, glutamic acid, aspartic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, arylsulfonic acid (e.g. p-toluene Sulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid or naphthalenedisulfonic acid), salicylic acid, glutaric acid, gluconic acid, tricavallylic acid, mandelic acid, cinnamic acid, substituted cinnamic acid (e.g. 4-methyl and 4- Methyl, methoxy, halo or phenyl substituted cinnamic acid, including methoxycinnamic acid and α-phenyl cinnamic acid), ascorbic acid, oleic acid, naphthoic acid, hydroxynaphthoic acid (eg, 1- or 3-hydroxy Roxy-2-naphthoic acid), naphthal Acrylic acid (eg, naphthalene-2-acrylic acid), benzoic acid, 4-methoxybenzoic acid, 2 or 4-hydroxybenzoic acid, 4-chlorobenzoic acid, 4-phenylbenzoic acid, benzeneacrylic acid (eg, 1,4- Benzenediacrylic acid) and isethionic acid. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts and organic bases such as dicyclohexylamine and N -methyl- D -Salts with glucamine.

The compounds according to the invention also have the potential of combining long lasting effects and rapid onset of action. Certain compounds also exhibit improved therapeutic indicators in animal models compared to existing long-lasting PDE-4 inhibitors. In addition, the compounds of the present invention may be suitable for once to three times daily administration.

The amount of the compound of formula (1) or a pharmaceutically acceptable salt thereof used to achieve a therapeutic effect, as well as the particular compound, the method of administration, the subject to be treated, and the tablet disorder or disease to be treated, may range from 0.0005 mg to 10 mg. mg, preferably orally, or inhaled at a dose of 0.005 mg to 5 mg. Dosage ranges for adults are generally from 0.0005 mg to 100 mg per day, preferably from 0.01 mg to 5 mg per day.

It is possible to administer the compound of formula (1) or a pharmaceutically acceptable salt thereof according to the invention alone, but preferably in a pharmaceutical formulation. Accordingly, the present invention is also preferably provided as a pharmaceutical formulation comprising Formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, and any one or more therapeutic ingredients.

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

Example 1 Preparation of 1- (1- (3- ( cyclopropylmethoxy ) -4- ( difluoromethoxy ) phenyl) ethyl) -3-phenyl-1 H- pyrazole (Compound 10)

1- (3- ( cyclopropylmethoxy )-4-( difluoromethoxy ) phenyl ) ethanol Manufacture

3.0 g (12.4 mmol) of 3- (cyclopropylmethoxy) -4- (difluoromethoxy) benzaldehyde was dissolved in 30 mL of dried ethyl ether, cooled to 0 ° C. under a nitrogen stream, and then 6.2 mL (18.6) of MeMgBr (3.0 M / Et ₂ O). mmol) was added slowly and stirred at 0 ° C. for 1 hour. 30 mL of water was added to the reaction solution to terminate the reaction. The mixture was neutralized with 1N-HCl solution to pH 4 and extracted with ethyl acetate (100 mLx2), followed by washing with water and brine. The organic layer was dried over anhydrous sodium sulfate, filtered and the residue obtained by distillation of the solvent under reduced pressure was purified by silica gel column chromatography (n-hexane / ethyl acetate = 3/1) to give 2.61 g (yield 81%) of the title compound as a colorless oil. Got.

¹ H NMR (300 MHz, CDCl ₃ ) δ 0.32 (m, 2H), 0.62 (m, 2H), 1.27 (m, 1H), 2.28 (brs, 1H), 3.85 (d, 2H), 4.80 (q, 1H), 6.59 (t, 1H, J = 75.7 Hz, CHF ₂ ), 6.86 (m, 1H), 6.97 (1H, d, J = 1.8 Hz), 7.07 (d, 1H, J = 8.1 Hz)

Preparation of 4- (chloromethyl) -2- (cyclopropylmethoxy) -1- (difluoromethoxy) benzene

Dissolve 2.5 g (9.7 mmol) of prepared 1- (3- (cyclopropylmethoxy) -4- (difluoromethoxy) phenyl) ethanol in 10 mL of dry benzene, add 1.4 mL (19.4 mmol) of SOCl ₂ , and heat to 70 ° C for 1 hour. Reacted. The reaction solution was concentrated under reduced pressure, and then purified by silica gel column chromatography (n-hexane / ethyl acetate = 15/1) to obtain 2.2 g (yield 82%) of the title compound as a colorless oil.

¹ H NMR (300 MHz, CDCl ₃ ) δ 0.34 (m, 2H), 0.64 (m, 2H), 1.27 (m, 1H), 1.81 (d, 3H) 3.88 (d, 2H), 5.02 (q, 1H ), 6.61 (1H, t, J = 75.5 Hz, -CHF ₂ ), 6.93 (1H, dd, J = 2.0, 8.2 Hz), 7.01 (1H, d, J = 2.0 Hz), 7.11 (1H, d, J = 8.2 Hz)

Preparation of 1- (1- (3- (cyclopropylmethoxy) -4- (difluoromethoxy) phenyl) ethyl) -3-phenyl-1H-pyrazole (Compound 10)

300 mg (2.08 mmol) of 4- (chloromethyl) -2- (cyclopropylmethoxy) -1- (difluoro methoxy) benzene ( 2a ) was dissolved in 5 mL of DMF, followed by 365 mg (1.39 mmol) of cesium and 3-phenylpyrazole. 1.36 g (4.16 mmol) of carbonate were added and stirred at room temperature for 15 hours. 50 mL of water was added to the reaction mixture, the mixture was diluted with ethyl acetate (60 ml x 2), and washed with water and brine. The organic layer was dried over anhydrous sodium sulfate, filtered and the solvent was distilled under reduced pressure, and the residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 3/1) to give 600 mg (yield 75%) of a colorless oil compound 10. Got it.

¹ H NMR (300 MHz, CDCl ₃ ) δ 0.31 (m, 2H), 0.59 (m, 2H), 1.22 (m, 1H), 1.91 (d, 3H) 3.80 (d, 2H), 5.53 (q, 1H ), 6.33 (t, 1H, CHF ₂ , J = 75.5 Hz), 6.29 (d, 1H, J = 2.3 Hz), 6.80-6.85 (m, 2H), 7.10 (d, 1H, J = 8.1 Hz), 7.29 (d, 1H, J = 7.4 Hz), 7.36-7.41 (m, 3H), 7.82 (d, 2H, J = 8.6 Hz)

The structure and ¹ H NMR of the compounds prepared in the same manner as shown in Table 1 below.

TABLE 1

Example 2

PDE -4 active Inhibitory ability  analysis

Inhibitory activity of PDE-4d was performed by modifying the method of Kate et al. (Kate, et al., JBC , 271: 796 (1996)). PDE-4d used human PDE-4d prepared by genetic recombination technology.

The PDE4d gene was cloned into the GST fusion expression vector pGEX4T3 (APBiotech), and then transformed into E. coli BL21, DE3. After incubating the strain at 37 ° C. in LB medium, the temperature was lowered to 18 ° C. when 0.3-0.5 OD was induced to induce PDE4d expression with 0.5 mM IPTG. After 15 hours of expression induction, PDE4d-expressing cells were obtained, and the cells were suspended in a buffer solution (50 mM Tris, pH 8.0, 0.4 M NaCl, 5 mM DTT), disrupted, and purified by centrifugation. The columns used for purification are Q-Sepharose (APBiotech), Glutanion-Sepharose (APBiotech), and Superdex 200 gel filtration chromatography (APBiotech) in this order.

In order to measure the ability to inhibit PDE4d activity of each compound, 1 nM of PDE4d and 250 nM of cAMP (Sigma) as a substrate were added to the reaction buffer (50 mM Tris-HCl, pH 7.5) and 4 mM MgCl ₂ , and obtained in Example 1 above. Each compound was added at a concentration of 1 to 50 uM and then reacted at 34 ° C. for 45 minutes. The reaction total was 60 µl. At this time, cAMP was used by mixing a ratio of [ ³ H] cAMP (Amersham, 1 μCi / μl) and cAMP (hot: cold, that is, isotope: non-isotope) at 1: 200. The reaction was then stopped by heating at 95 ° C. for 2 minutes, then cooled for 3 minutes and 30 μl of 1 mg / ml Snake Venome (sigma V0376) was added. Subsequently, after 30 minutes of reaction at 34 ° C, 30 μl of the reaction solution was poured into 250 μl of DOWEX 1 × 2-100 ion exchange resin (Aldrich, pre-mixed at a ratio of 1 to 1.6 of distilled water) and stirred vigorously for 2 minutes. Then, the resin was allowed to settle, and 130 μl of the supernatant was added to 2 ml of scintillation cocktail (Packard), mixed well, and measured by beta-counter. The concentration of each test compound that inhibited 50% of the enzyme activity compared to the enzyme activity of the control group without the addition of the compound of Formula 1 was determined as IC ₅₀ .

Example 3

PDE -4 Enzyme Selectivity Assay

In order to measure the enzyme selectivity against PDE4d, the enzyme activity inhibitory effect of each test compound against PDE-3a, PDE-5a, and PDE-7a was measured. PDE3a and PDE7a, which used cAMP as a substrate, measured the degree of activity inhibition using the Amplified Luminescent Proximity Homogeneous Assay (Alpha) Screen method, and PDE5a, which used cGMP as a substrate, used the enzyme of each test compound by FP (fluorescence polarization) method. Activity inhibition efficacy was measured.

PDE3a, PDE5a and PDE7a all used human enzymes prepared by genetic recombination techniques.

In order to measure the inhibitory activity of PDE3a and PDE7a of each test compound, first put each compound (final DMSO 1%) at the concentration to be tested in a 96 well half area white plate and follow the same conditions as the conditions for measuring PDE4d enzyme activity described above. Enzyme reaction at When measuring enzyme activity using Alpha Screen, the concentrations of PDE3a and PDE7a should be 10nM and 80nM, respectively, and were carried out according to the method proposed in the Alphascreen cAMP assay kit (Perkin Elmer life science). Briefly, after the completion of the reaction of PDE3a and PDE7a, a final 10 nM biotinylated-cAMP and 20ul / mL donor beads are added to the reaction mixture. The donor bead is sensitive to light, so be careful not to expose it to light and let it react for 30 minutes at room temperature. Then add acceptor beads to a final 20ul / ml. After reacting for 45 minutes at room temperature, the reaction was further reacted for 20 minutes in the Fusion Alpha-FP device, and the emission at 520-620 nm was measured.

In the enzymatic reaction of PDE3a and PDE7a, the signal increase and decrease is caused by the mutual competition between the remaining cAMP and the newly added biotinylated-cAMP, which is not used as a substrate. When the enzyme activity of PDE3a and PDE7a is inhibited by each test compound, signal is reduced.

PDE5a, which uses cGMP as an enzyme substrate, measured the enzyme inhibitory ability of each test compound according to the experimental method presented by IMAP ^TM Phosphodiesterase Assay Kit (Molecular Device), which measures enzyme activity by fluorescence polarization (FP) method. First, add each test compound (final DMSO 5%) at the concentration to be tested in 96 well black plate, and add FL-cGMP (Fluorescence-labeled derivatives of cGMP) and final 200 nM PDE5a. After reacting for 45 minutes at 30 degrees, add the IMAP binding reagent provided by the kit. After reacting at room temperature for 60 minutes, FP obtained by 485 nm exitation and 535 nm emission is measured. The more enzyme activity of PDE5a, the higher the FP signal. The inhibition of enzyme activity of PDE5a by each test compound can be determined by measuring the decreased FP signal.

Example 4

In vivo Asthma Treatment Test

For in vivo animal experiments, 8 to 9 week old male C57BL / 6J mice weighing 20-25 g were used as experimental animals. For sensitization and challenge of airway and lung by ovalbumin (OVA), OVA mixed with Al (OH) ₃ was firstly sensitized by intraperitoneal administration to mice. 2nd sensitization. Six days after the second sensitization, 5% OVA was inhaled through the nebulizer for three days. Drug administration to sensitized mice was administered orally (po) three times each 1 hour prior to inhalation of the OVA. 0.5% CMC was administered as a control. To observe the activity and cell composition of the mouse tracheal alveolar lavage fluid, the mice were anesthetized after 62 hours, inoculated with OVA, the cervical thorax was opened, the tube was inserted into the trachea using a tube containing 0.8 ml of PBS, and then PBS was injected. After massaging the chest for about 30 seconds, a cell suspension was obtained from the lungs. The obtained cells were centrifuged at 400xg, and the supernatant was used for measuring the eosinophil peroxidase activity (EPO activity), and pellets were used for cell composition investigation. To measure the activity of basophils, 100 ul of cell suspension was reacted with 0.1 mM OPD, 0.05 M Tris-Hcl (pH 8.0), Triton X-100 and 1 mM H ₂ O _2, and the absorbance was measured at 492 nm. . For staining of basophils, the cell pellet was resuspended in PBS and centrifuged at 150xg for 5 minutes to ensure that the cells adhered to the slides. The attached cells were subjected to Diff-Quick staining to observe the number of cells under a microscope. Airway hyper-responsiveness (AHR) analysis was performed using a whole body plethysmographer in living animals. 48 hours after OVA inhalation, Methacholine (Mch) was inhaled with nebulizer, and airway reactivity by Mch was measured using whole body plethysmography to compare enhanced pause (Penh) values for respiratory ability of airways.

Example 5

Emesis  Related Side Effect Test Methods

Ferret (Male) purchased from Marshal (1.5-2kg) was divided into Control, Yohimbine, # 423-500mg / kg, and # 423-1000mg / kg treatment groups. The drug dissolved in% CMC was orally administered. Thereafter, visual observation was performed for 3 hours to determine the number of vomitting, salivation and diarrhea in each animal.

IC ₅₀ values for the PDE-4 enzymes of the compounds are shown in Table 2 below.

TABLE 2

As can be seen in Table 2, R ₁ of the compound of Formula 1 showing PDE-4 inhibitory activity is fixed with a compound substituted with a difluoromethoxy group, and the activity changes depending on R ₂ , but the difluoromethoxy group The compound substituted with a cyclopropylmethyl group showed a tendency of better PDE-4 inhibitory activity than the compound substituted with (19, 20, 21, 22) or the compound substituted with a cyclopropyl group (31, 32). R ₃ is a compound substituted with a phenyl group is good activity, a compound substituted with a meta-substituted phenyl group shows a better activity than a phenyl group substituted in the para position (2 and 12, 3 and 102, 6 and 24), pyridine or Compounds substituted with furan, thiophene and the like maintained good activity (1, 8, 103-111). Compounds with substituents larger than hydrogen in R ₅ have reduced activity (9, 11, 13, 14, 25, 36, 37), and compounds with substituents larger than methyl in R ₅ have slightly decreased activity (14, 16, 59, 60, 61, 64, 67-71, 76-78, 89, 90, 94, 96, etc.) Compounds substituted with carboxylic acid or amide groups tended to be less active (62, 63, 65). 73, 86-88, 91-93, 95, etc.). In total, R ₁ is difluoromethyl, R ₂ is cyclopropylmethyl, R ₄ and R ₅ are hydrogen and R ₆ is a methyl group, R ₃ is phenyl as tetrazole, nitro, derivative of carboxylic acid and meta _{In the case of trivalent} pyridine (Compounds 1, 3, 6, 34, 40, 45), PDE-4 inhibitory activity was the best.

In vivo asthma treatment efficacy test results (AHR and EPO) using asthma model animals by selecting compound 7, compound 8 and compound 115 in consideration of water solubility, logP and pharmacokinetics among the active compounds (AHR and EPO) 100 mg / kg oral At doses, asthma was similar to Roflumilast (30 mg / kg). In addition, Compound 115 showed a significant therapeutic effect on arthritis in the animal model of CIA mice at the oral dose of 100 mg / kg.

As a result of vomiting-related side effects of representative compound 115, the average amount of vomiting was observed in all the ferrets in the yohimbine-treated group (7 mg / kg). The animals with saliva and diarrhea were 66 Was%. On the contrary, no abnormal symptoms were observed at the 500 mg / kg and 1,000 mg / kg treatment concentrations.

Formula 1 compounds according to the present invention as a novel compound has excellent activity for PDE-4 enzymes and high selectivity for PDE isozyme and shows that the asthma treatment effect in animal experiments is very good. In addition, the compound series has improved the vomiting-related side effects, which is the most serious problem of PDE-4 inhibitors, as well as asthma and chronic obstructive pulmonary diseases, as well as inflammatory diseases including arthritis and atopic dermatitis, and brain diseases such as Alzheimer's, depression and memory loss. It is expected to have an excellent effect on prophylactic and therapeutic agents and various cancer treatments including leukemia, and at the same time, there are no side effects.

Formula 1 compounds according to the present invention are novel compounds with excellent activity against PDE-4 enzymes and high selectivity for other PDE enzymes, which can be used in animal studies to treat bronchial asthma, COPD treatment, asthma and chronic obstructive pulmonary disease. It is expected to have an excellent effect in the treatment of diseases related to inflammatory diseases including Chronic Obstructive Pulmonary Disease, and diseases of the central nervous system such as arthritis, atopic dermatitis, leukemia and degenerative brain diseases such as various cancers and Alzheimer's disease. There is little advantage.

Claims (14)

The novel 1- [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof. [Formula 1]

[In Formula 1, R ₁ is straight chain or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl or halogen substituted straight chain or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl; R ₂ is straight or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl, substituted by halogen straight or branched saturated or unsaturated (C ₁ -C ₇ ) alkyl, (C _3- C ₇ ) cycloalkyl group or (C ₃ -C ₈ ) cycloalkylmethyl; R _3, R ₄ and R ₅ independently of one another are hydrogen, phenyl, pyridinyl, N-oxypyridinyl, straight or branched saturated and unsaturated (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cyclo Alkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, halogen atom, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino, mono or di (C ₁ -C ₇ ) alkylaminocarbonyl, (C ₃ -C ₇ ) cycloalkylamino, morpholino, piperazinyl, furanyl, thiophenyl, guanidinyl, ureido, benzyl, benzyloxy, formyl, (C ₁ - C ₇₎ alkanoyl, (C ₁ - C ₇₎ alkoxy, (C ₁ - C ₇₎ alkoxy (C ₁ - C ₇₎ alkyl, hydroxy (C ₁ - C ₇₎ alkyl, carboxyl, (C ₁ C ₇ ) alkoxycarbonyl, wherein said (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, phenyl, pyridinyl and N-oxypyridinyl are (C ₁ -C ₇ ) alkyl groups, (C ₃ -C ₇ ) cycloalkyl groups, (C ₄ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group, (C ₁ -C ₇ ) alkoxy, tetrazolyl, morpholino, N-oxymorpholino, piperazinyl, N-oxypiperazinyl, Selected from furanyl, thiophenyl, pyrrolidinyl, piperidinyl, pyridinyl, pyrimidinyl, halogen, nitro, cyano, hydroxy, carboxyl or amino groups, mono or di (C ₁ -C ₇ ) alkylamino One or more may be further substituted; R ₆ is straight or branched saturated and unsaturated (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₃ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl, phenyl, phenoxy, benzyl, benzyloxy, (C ₁ -C ₇ ) alkoxy, (C ₁ -C ₇ ) alkoxy (C ₁ -C ₇ ) alkyl, alkyl, cycloalkyl of R ₆ , The phenyl group is a (C ₁ -C ₇ ) alkyl group, (C ₃ -C ₇ ) cycloalkyl group, (C ₄ -C ₁₀ ) cycloalkyl (C ₁ -C ₇ ) alkyl group, (C ₁ -C ₇ ) alkoxy, halogen, One or more selected from nitro, cyano, hydroxy, carboxyl or amino groups, mono or di (C ₁ -C ₇ ) alkylamino may be further substituted, provided that R _3, R ₄ and R ₅ are all hydrogen Is excluded.] delete The method of claim 1, R ₁ and R ₂ are each independently of each other methyl, difluoromethyl, cyclopentyl or cyclopropylmethyl 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole Compounds, or pharmaceutically acceptable salts thereof. The method of claim 3, wherein R ₃ is cyclopentyl, cyclohexyl, phenyl, pyridinyl, N-oxypyridinyl, morpholino, piperazinyl, furanyl, or thiophenyl, wherein said cyclopentyl, cyclohexyl, phenyl and pyridinyl or N Oxypyridinyl is hydroxy, Halogen, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino, (C ₁ -C ₇ ) alkoxy (C ₁ -C ₇ ) alkyl, (C ₃ -C ₇ ) cycloalkylamino , Morpholino, N-oxymorpholino, N-oxypiperazinyl, piperazinyl, furanyl, thiophenyl, guanidinyl, ureido, carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl , Can be substituted with carbamoyl; R ₄ and R ₅ are independently of each other a hydrogen atom, (C ₁ -C ₇ ) alkyl or phenyl, said phenyl may be further substituted with carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl, carbamoyl Characterized 1- [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole compound, or a pharmaceutically acceptable salt thereof. The method of claim 1, R ₆ is a (C ₁ -C ₇ ) alkyl group or phenyl, the alkyl group being carboxyl, (C ₁ -C ₅ ) alkoxycarbonyl, carbamoyl, hydroxy, 1- [1- (), characterized in that one or more selected from halogen, cyano, nitro, amino, mono- or di (C ₁ -C ₇ ) alkylamino or (C ₁ -C ₇ ) alkoxy can be substituted 3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound, or a pharmaceutically acceptable salt thereof. The method of claim 1, R _3, R ₄ and R ₅ are 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compounds, or pharmaceutically acceptable thereof, wherein they are selected from the following substituents salt. 3-pyridinyl, N-oxypyridin-3-yl 3- (ethoxycarbonyl) phenyl 3-carboxyphenyl 4-pyridinyl 4- (acetamido) phenyl N-oxypyridin-4-yl 3-aminophenyl 3- (benzyloxy) phenyl 3-hydroxyphenyl 3-((ethoxycarbonyl) methoxy) phenyl 3- (benzamido) phenyl 3- (methoxy) phenyl 3- (dibenzylamino) phenyl 3- (benzylamino) phenyl 3- (methylamino) phenyl 3- (2- (N, N-dimethylaminoethoxy) phenyl 3- (dimethylamino) phenyl 3-chlorophenyl 3-bromophenyl 3- (pyrrolidin-1-yl) phenyl 3- (piperidin-1-yl) phenyl Phenyl 3- (methanesulfonylamino) phenyl 3-((N, N-di- (methanesulfonyl)) amino) phenyl 4- (benzyloxy) phenyl 3-cyanophenyl 3- (2H-tetrazol-5-yl) phenyl 4-hydroxyphenyl 4-((ethoxycarbonyl) methoxy) phenyl 4-methoxyphenyl 3- (i-propoxycarbonyl) phenyl 4- (carboxymethoxy) phenyl 3- (trifluoromethanesulfonylamino) methyl 3-((N, N-di- (trifluoromethanesulfonyl) amino) methyl 4- (ethoxycarbonyl) phenyl 4-carboxyphenyl 4- (2-chloropyridinyl) phenyl 4-cyanophenyl 4- ( 1H -tetrazol-5-yl) phenyl 3,5-dichloropyridin-4-yl 4- (N-oxypyridinyl) 4-nitrophenyl 3-amino-4-chlorophenyl 4-aminophenyl 3-cyano-4-fluorophenyl 3-bromo-4-methoxyphenyl 3,5-dibromophenyl (3-carboxy-4-methoxy) phenyl 3-ethylaminophenyl (3-amino-4-methyl) phenyl (3-nitro-4-methyl) phenyl (3-ethoxycarbonyl-4-methyl) phenyl (3-carboxy-4-methyl) phenyl 3-ureidophenyl 3-((ethoxycarbonyl) methylamino) phenyl 3-((carboxy) methylamino) phenyl 3-morpholinophenyl 3- (tetrahydro-2,4,6-trioxopyrimidin-1 (2H) -yl) phenyl 3-((ethoxycarbonylmethyl) carbamoyl) phenyl 3- (1- (ethoxycarboxy) -2-phenylethylcarbamoyl) phenyl 3-carbamoylphenyl (3-carboxy-4-fluoro) phenyl Cyclohexyl 3-cyanocyclohexyl 3-ethoxycarbonylphenyl 3-carboxycyclohexyl 4-ethoxycarbonylphenyl Morpholino 4- (4-toluenesulfonyl) piperazin-1-yl 3-((1-methoxycarbonyl-2-methyl) butylcarbamoyl) phenyl 3-((1-methoxycarbonyl) ethylcarbamoyl) phenyl 3-((1-methoxycarbonyl-2-methyl) propylcarbamoyl) phenyl 3-((1-carboxy-2-methyl) propylcarbamoyl) phenyl 3-((1-carboxy-3-methyl) butylcarbamoyl) phenyl 3-((1-carboxy) ethylcarbamoyl) phenyl 3-((2,2,2-trifluoroethoxy) carbonyl) phenyl 3-((1- (propyl-2-oxycarbonyl) -2-methyl) propylcarbamoyl) phenyl 3-((1-ethoxycarbonyl-3-methyl) butylcarbamoyl) phenyl 3-((1- (propyl-2-oxycarbonyl) -3-methyl) butylcarbamoyl) phenyl 3-((pivaloyloxy) methoxycarbonyl) phenyl 3-chloro- (N-oxypyridin-4-yl) Furan-2-yl Furan-3-yl 5-methylfuran-2-yl 3-chlorofuran-2-yl 5-chlorofuran-2-yl Thiophen-2-yl Thiophen-3-yl 3-methylthiophen-2-yl 5-methylthiophen-2-yl 3-chlorothiophen-2-yl 5-chlorothiophen-2-yl 5-bromothiophen-2-yl 2,5-dichlorothiophen-3-yl The method of claim 6, R ₆ is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, ethoxycarbonylmethyl, carboxymethyl, hydroxymethyl, 2-hydroxyethyl, cyanomethyl or carbamoylmethyl 1- [1- (3,4-dialkoxyphenyl) -alkyl] -1 H -pyrazole compound, or a pharmaceutically acceptable salt thereof. The method of claim 7, wherein 1- (dialkoxybenzyl) -1H-pyrazole compound, or a pharmaceutically acceptable salt thereof, characterized in that it is selected from the following compounds:

[In the compound, R ₆ is selected from methyl, ethyl, ethoxycarbonylmethyl, carboxymethyl, cyanomethyl.] A process for preparing the compound of formula 1 according to claim 1 by reaction of a halobenzyl compound represented by formula (2) with a pyrazole compound represented by formula (3).

Asthma or chronic obstructive pulmonary disease (Chronic Obstructive Pulmonary Disease) containing 1- [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole derivative represented by Formula 1 as an active ingredient Pharmaceutical compositions for the treatment or prophylaxis. delete Treatment or prevention of arthritis, atopic dermatitis, leukemia or cancer containing 1- [1- (3,4-diakoxyphenyl) -alkyl] -1 H -pyrazole derivative represented by the formula (1) as an active ingredient Pharmaceutical Compositions For. The treatment and prevention of degenerative brain disease or improvement of memory, comprising 1- [1- (3,4-alkoxyphenyl) -alkyl] -1 H -pyrazole derivative represented by the formula (1) as an active ingredient. Pharmaceutical composition for. The method of claim 13, Degenerative brain disease is Alzheimer's, dementia, characterized in that the pharmaceutical composition.