KR20170046847A

KR20170046847A - Novel N-aryl-1H-pyrazolopyridin-3-amine derivatives or pharmaceutically acceptable salts thereof, preparation method therof and pharmaceutical composition for use in preventing or treating MELK(maternal embryonic leucine zipper kinase) activity related diseases containing the same as an active ingredient

Info

Publication number: KR20170046847A
Application number: KR1020150146653A
Authority: KR
Inventors: 최상운; 박철민; 이선경; 송종환; 박지훈; 손유화; 김광록; 김지은
Original assignee: 한국화학연구원
Priority date: 2015-10-21
Filing date: 2015-10-21
Publication date: 2017-05-04
Also published as: KR101753652B1

Abstract

The present invention relate to N-aryl-1H-pyrazolopyridin-3-amine derivatives or pharmaceutically acceptable salts thereof, a preparation method thereof and a pharmaceutical composition for preventing or treating maternal embryonic leucine zipper kinase (MELK) related diseases containing the same as an active ingredient. The N-aryl-1H-pyrazolopyridin-3-amine derivatives according to the present invention has an excellent effect of inhibiting the expression of MELK, thereby being usefully used for preventing or treating MELK-related diseases, such as breast cancer, lung cancer, bladder cancer, lymphoma, glioblastoma multiforme, and cervical cancer.

Description

N-aryl-1H-pyrazolopyridine-3-amine derivative or a pharmaceutically acceptable salt thereof, a process for producing the same, and a pharmaceutical composition for preventing or treating a MELK- 1H-pyrazolopyridin-3-amine derivatives or their salts, preparation method, and pharmaceutical composition for use in preventing or treating MELK (maternal embryonic leucine zipper kinase)

The present invention relates to an N-aryl-1H-pyrazolopyridin-3-amine derivative or a pharmaceutically acceptable salt thereof, a process for the preparation thereof, and a method for the prevention or treatment of MELK (maternal embryonic leucine zipper kinase) &Lt; / RTI >

MELK (Maternal Embryonic Leucine zipper Kinase) is a serine / threonine kinase that is activated by adenosine monophosphate belonging to the snf1 / AMPK family involved in mammalian embryonic development and contains vertebrate animal cells It is known that the amount or activation of the cell cycle is maximized during the mitotic phase of the cell cycle.

It is also known that MELK is expressed in stem cells and progenitor cells of adult tissues associated with various biological processes and plays an important role in stem cell regeneration, cell cycle and pre-mRNA splicing, etc. have. CDC25, which is involved in the cell cycle, is known to be phosphorylated by MELK. It is known that the protein regulates the cell cycle by phosphorylating the transcription protein ZPR9. In particular, MELK is known to play a role as a cell cycle regulator in tumor cell lines. Recent analysis of the gene expression profile using a genome wide cDNA microarray containing 23,040 genes confirmed that MELK was upregulated in breast cancer In addition, MELK is overexpressed in Glioblastoma multiforme (GBM), a type of brain tumor, which is overexpressed in glioma stem cells (GSCs), especially in the glia tissue. MELK has been implicated in the growth and survival of glial stem cells and has been shown to be upregulated in cancer cells of various types of cancer, such as lung, bladder, lymphoma, cervical cancer, .

Recently, studies using cancer models have shown that reducing the expression of MELK by RNA interference or a low molecular inhibitor leads to the death of cancer stem cells (CSCs) derived from polymorphonalephritis and breast cancer It turned out.

On the other hand, although the activation mechanism of MELK is not limited, it is known that it directly binds to tumor gene transcription factors c-JUN and FOXM1 in cancer cells but not in normal cells. That is, MELK is not expressed in normal organs (heart, liver, lung, and kidney), and is overexpressed in breast cancer, polymorphic hypertrophy, and a majority of multiple tumor cell lines.

As described above, MELK is not expressed in normal cells, but is expressed specifically in tumor cells. As the expression of MELK is decreased, cancer stem cell death is induced, thereby suppressing the expression of MELK Is the target of cancer treatment because it can inhibit the growth of cancer cells alone. Currently, there is OTSSP167, which is developed in 2013 and is in clinical phase 1, as a drug under development.

Accordingly, the present inventors have found that N-aryl-1H-pyrazolopyridin-3-amine derivatives or pharmaceutically acceptable salts thereof are excellent in inhibiting the expression of MELK, and thus can be used for the treatment of breast cancer, lung cancer, bladder cancer, lymphoma, , Cervical cancer, and the like, and completed the present invention.

Molecular < / RTI > Cancer (2014) 13,100, pp1-14 Physiological Reports (2014) 2 (9) PLoS One (2014) 9 (4), e92546 / 1-10 Molecular Cancer Therapeutics (2014) 13 (6), 1393

It is an object of the present invention to provide an N-aryl-1H-pyrazolopyridin-3-amine derivative or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a process for preparing the N-aryl-1H-pyrazolopyridin-3-amine derivative.

Still another object of the present invention is to provide a pharmaceutical composition for preventing or treating MELK-related diseases containing the above-mentioned N-aryl-1H-pyrazolopyridin-3-amine derivative, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient To provide a composition.

Another object of the present invention is to provide a method for preventing or ameliorating a MELK-related disease comprising the N-aryl-1H-pyrazolopyridin-3-amine derivative, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient, To provide a composition.

In order to achieve the above object,

The present invention provides a compound represented by the following general formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In Formula 1,

X, Y and Z are each independently carbon or nitrogen;

HetAr is 5- to 8-membered heteroarylene comprising at least one heteroatom selected from the group consisting of N, O and S;

R ¹ is hydrogen, straight or branched C _1- ₅ alkyl or unsubstituted N, O and S, and 5-8 membered heterocycloalkyl of atoms including a hetero atom at least one member selected from the group consisting of;

R ² is hydrogen or unsubstituted or substituted C _6- ₁₀ aryl, wherein the substituted C _6- ₁₀ aryl is optionally substituted with one or more substituents selected from the group consisting of C _1- ₅ alkoxy halogen and straight-chain or branched &Lt; / RTI >

R ³ is one or more substituents selected from hydrogen or straight chain or C _1- ₅ alkyl, C _1- ₅ alkoxy and the group consisting of halogen in the side chain;

R ⁴ is hydrogen or linear or branched C _1- ₅ alkyl; And

n is an integer of 0-5.

Also, as shown in the following Reaction Scheme 1,

Reacting a compound represented by the formula (2) with a compound represented by the formula (3) to obtain a compound represented by the formula (4) (step 1);

A step of reacting a compound represented by the formula (4) obtained in the above step 1 with a compound represented by the formula (5) to obtain a compound represented by the formula (1) (step 2) :

[Reaction Scheme 1]

In the above Reaction Scheme 1,

A ¹ is fluoro, chloro or bromo, A ² is chloro, bromo or iodo, A ¹ is fluoro, A ² is chloro, bromo or iodo and A ¹ is chloro, A ² is bromo or iodo; when A ¹ is bromo, A ² is iodo;

D is

or

ego; And

X, Y, Z, R ¹ , R ² , R ³ , R ⁴ , HetAr and n are the same as defined in Formula 1.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating a MELK-related disease, which comprises the compound represented by the above-mentioned formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating a MELK-related disease, which comprises the compound represented by the formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

The N-aryl-1H-pyrazolopyridine-3-amine derivative according to the present invention is excellent in the effect of suppressing the expression of MELK, and thus can be used as a MELK such as breast cancer, lung cancer, bladder cancer, lymphoma, And can be useful for preventing or treating a related disease.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following general formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

X, Y and Z are each independently carbon or nitrogen;

R ⁴ is hydrogen or linear or branched C _1- ₅ alkyl; And

n is an integer of 0-5.

Preferably,

X, Y and Z are each independently carbon or nitrogen;

HetAr is a 5-6 membered heteroarylene comprising at least one N atom;

R ¹ is a heterocycloalkyl of 5 to 6 atoms containing one or more C _1- ₃ alkyl or an unsubstituted N atom of hydrogen, linear or branched;

R ² is hydrogen or an unsubstituted or substituted _6- C ₈ aryl, wherein the substituted _6- C ₈ aryl is substituted with one or more substituents selected from the group consisting of C _1- ₃ alkoxy and halogen straight-chain or branched &Lt; / RTI >

R ³ is one or more substituents selected from hydrogen or straight chain or C _1- ₃ alkyl or C _1- ₃ alkoxy and the group consisting of halogen in the side chain;

R ⁴ is hydrogen or linear or branched C _1- ₃ alkyl; And

n is an integer of 1-3.

More preferably,

X, Y and Z are each independently carbon or nitrogen;

HetAr is pyrrolylene, pyrazolylene, pyridinylene or pyrazinylene;

R ¹ is hydrogen, methyl, ethyl, propyl, pyrrolidinyl or piperidinyl;

R ² is hydrogen or unsubstituted or substituted phenyl, wherein said substituted phenyl may be substituted with one or more substituents selected from the group consisting of fluoro, chloro, bromo, methoxy and ethoxy;

R ³ is hydrogen or one or more substituents selected from the group consisting of fluoro, chloro, bromo, methyl, ethyl, methoxy and ethoxy;

R < ⁴ > is hydrogen, methyl, ethyl or propyl; And

n is an integer of 1-2.

Most preferably,

X, Y and Z are each independently carbon or nitrogen;

R ¹ -HetAr is

,

,

,

or

ego;

R < ¹ > is hydrogen, methyl or

ego;

R ² is hydrogen,

or

ego;

R ³ is hydrogen or one or more substituents selected from the group consisting of fluoro, chloro, methyl and methoxy;

R < ⁴ > is hydrogen or methyl; And

n is 1.

Preferable examples of the compound represented by the formula (1) according to the present invention include the following compounds:

(1) 1- (2-Chlorobenzyl) -N- (3-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(2) l- (2-Chlorobenzyl) -N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(3) l- (2-Chlorobenzyl) -N- (2-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(4) Synthesis of 1- (2-chlorobenzyl) -N- (2,3-dichlorophenyl) -5- (pyridin- ;

(5) l- (2-Chlorobenzyl) -N- (3-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(6) l- (2-Chlorobenzyl) -N- (4-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(7) 1- (2-chlorobenzyl) -N- (5-fluoro-2-methylphenyl) Amine;

(8) l- (2-Chlorobenzyl) -N- (2,5-dimethylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(9) 1- (2-Chlorobenzyl) -N- (3,5-dimethoxyphenyl) -5- (pyridin- 3- yl) -lH- pyrazolo [3,4- b] Amine;

(10) 1- (2-Chlorobenzyl) -N- (4-chlorophenyl) -N-methyl-5- (pyridin- Amine;

(11) l- (2-Chlorobenzyl) -N- (pyridin-2-yl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(12) 1- (3-Methoxybenzyl) -N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(13) N- (2-Chlorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(14) N- (3-Chlorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(15) N- (3-Fluorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(16) N- (5-fluoro-2-methylphenyl) -1- (3-methoxybenzyl) -5- - amine;

(17) To a solution of N- (3-chlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol- 3-amine;

(18) N- (3-Chlorophenyl) -5- (1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(19) N- (3-Chlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(20) N- (3,4-Dichlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(21) N- (3,5-Dichlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(22) Synthesis of N- (3,5-dichlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol- ] Pyridine-3-amine;

(23) N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(24) N- (2-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(25) N- (3-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(26) N- (3-Fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(27) N- (4-Fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(28) N- (5-Fluoro-2-methylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(29) N- (2,5-Dimethylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;

(30) N- (3-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-c] pyridin-3-amine;

(3-chlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol-4-yl) -lH- pyrazolo [3,4- c] pyridine- 3-amine;

(32) To a solution of N- (3,4-dichlorophenyl) -5- (1- (piperidin-4-yl) ] Pyridine-3-amine;

(33) To a solution of N- (3,5-dichlorophenyl) -5- (1- (piperidin-4-yl) -1H-pyrazol- ] Pyridine-3-amine;

(34) N- (3-Chlorophenyl) -5- (lH-pyrazol-4-yl) -lH-pyrazolo [3,4-c] pyridine-3-amine.

The compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid and the like, aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, But are not limited to, but are not limited to, but are not limited to, but are not limited to, but are not limited to, halides, halides, halides, halides, halides, halides, But are not limited to, lactose, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene Sulfonates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, and the like, as well as sulfonates such as benzyl sulfonate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, -Sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving a derivative of the formula (1) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, Followed by filtration and drying. Alternatively, the solvent and excess acid may be distilled off under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

Furthermore, the present invention encompasses the compounds represented by the formula (1) and pharmaceutically acceptable salts thereof as well as solvates, optical isomers and hydrates thereof which can be prepared therefrom.

Also, as shown in the following Reaction Scheme 1,

A step of reacting a compound represented by the formula (4) obtained in the above step 1 with a compound represented by the formula (5) to obtain a compound represented by the formula (1) (step 2) .

[Reaction Scheme 1]

In the above Reaction Scheme 1,

D is

or

ego; And

Hereinafter, a method for preparing the compound represented by Formula 1 according to the present invention will be described in detail.

In the method for preparing the compound represented by the formula (1) according to the present invention, the step (1) is a step of reacting the compound represented by the formula (2) with the compound represented by the formula (3) to obtain the compound represented by the formula (4).

Specifically, A ² of the compound represented by the formula (2) and -NH ² of the compound represented by the formula (5) are reacted in the presence of a base and a palladium catalyst to obtain a compound represented by the formula (4).

Examples of the base include inorganic bases such as cesium carbonate, sodium t-butoxide, potassium t-butoxide, sodium hydroxide, sodium carbonate, potassium carbonate and sodium hydride; Or an organic base such as N, N-diisopropylethylamine (DIPEA), 1,8-diazabicyclo [5.4.0] -7-anesene (DBU), pyridine, triethylamine, Or may be used in excess, alone or in combination, and it is preferable to use cesium carbonate.

As the palladium catalyst, tris (dibenzylideneacetone) palladium (Pd ₂ (dba) ₃ ), tetrakis (triphenylphosphine) palladium (Pd (Ph ₃ P) ₄ ), palladium charcoal ), Bis (triphenylphosphine) palladium dichloride (PdCl ₂ (PPh ₃ ) ₂ ), [1,1-bis (diphenylphosphino) ferrocene] dichloropalladium (PdCl ₂ dimer _{([PdCl (allyl)] 2} ), palladium acetate (Pd (OAc) _2), palladium chloride (PdCl _2), and the like, tris (dibenzylideneacetone) palladium (Pd ₂ (dba) ₃₎ Is preferably used.

Further, reaction solvents which can be used include toluene, dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, acetonitrile; Lower alcohols including isopropanol, methanol, ethanol, propanol and butanol; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; These can be used singly or in combination, and toluene is preferably used.

In the process for preparing the compound represented by the formula 1 according to the present invention, the step 2 is a step of reacting the compound represented by the formula 4 obtained in the step 1 with the compound represented by the formula 5 to obtain the compound represented by the formula 1 .

Specifically, A ¹ of the compound represented by the formula (4) and boron of the compound represented by the formula (5) are reacted in the presence of a base and a palladium catalyst to obtain a compound represented by the formula (4).

Examples of the base include inorganic bases such as potassium carbonate, cesium carbonate, sodium t-butoxide, potassium t-butoxide, sodium hydroxide, sodium carbonate and sodium hydride; Or an organic base such as N, N-diisopropylethylamine (DIPEA), 1,8-diazabicyclo [5.4.0] -7-anesene (DBU), pyridine, triethylamine, Or they may be used in excess, alone or in combination, and it is preferable to use potassium carbonate.

In addition, as is the palladium catalyst [1,1-bis (diphenylphosphino) ferrocene] dichloropalladium (PdCl ₂ (dppf)), tris (dibenzylideneacetone) palladium (Pd ₂ (dba) _3), tetrakis (triphenylphosphine) palladium _{(Pd (Ph 3 P) 4} ), palladium charcoal (Pd-C), bis (triphenylphosphine) palladium dichloride _{(PdCl 2 (PPh 3) 2} ), allyl palladium chloride dimer ([PdCl (allyl)] _2), palladium acetate (Pd (OAc) _2), palladium chloride may be used. (PdCl _2), such as [1,1-bis (diphenylphosphino) ferrocene] dichloropalladium (PdCl ₂ (dppf)) is preferably used.

Further, reaction solvents which can be used include dimethylacetamide (DMA), toluene, dimethylformamide (DMF), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, acetonitrile; Lower alcohols including isopropanol, methanol, ethanol, propanol and butanol; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; , Which can be used singly or in combination, and it is preferable to use dimethylacetamide (DMA).

The starting material (the compound represented by the formula 2) Recipe 1

The compound represented by Formula 2, which is a starting material of Reaction Scheme 1,

Reacting a compound represented by the formula (6) with hydrazine hydrate to obtain a compound represented by the formula (7) (step 1);

Reacting a compound represented by the formula (7) obtained in the above step 1 to obtain a compound represented by the formula (8) (step 2);

Reacting the compound represented by the formula (8) with the compound represented by the formula (9) to obtain the compound represented by the formula (10) (step 3);

Reacting a compound represented by the formula (10) obtained in the above step 3 with a compound represented by the formula (11) to obtain a compound represented by the formula (12) (step 4); And

(Step 5) of reacting a compound represented by the formula (12) obtained in the step 4 with a compound represented by the formula (13) to obtain a compound represented by the formula (2):

[Reaction Scheme 2]

In the above Reaction Scheme 2,

A < ³ > is halogen;

L ¹ is a C _1- ₃ alkoxy or halogen straight-chain or branched; And

R ² and n are the same as defined in the above formula (1).

Hereinafter, a method for preparing the compound represented by Formula 2 according to the present invention will be described in detail.

In the process for preparing a compound represented by the general formula (2) according to the present invention, the step (1) is a step of reacting a compound represented by the general formula (6) with hydrazine hydrate to obtain a compound represented by the general formula (7).

Specifically, the compound represented by the formula (6) is obtained by reacting -CN and L ¹ of the compound represented by the formula (6) with hydrazine hydrate to form a pentane ring.

At this time, usable reaction solvents include lower alcohols including ethanol, butanol, isopropanol, methanol and propanol; Dimethylformamide (DMF), toluene, dimethylacetamide (DMA), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, acetonitrile; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; These may be used singly or in combination, preferably ethanol.

In the process for preparing a compound represented by the general formula (2) according to the present invention, the step (2) is a step of reacting a compound represented by the general formula (7) obtained in the step (1) to obtain a compound represented by the general formula (8).

Specifically, a step of reacting a compound represented by the formula (7) to remove -NH ₂ to obtain a compound represented by the formula (8).

At this time, the reaction solvent which can be used is a lower alcohol including methanol, ethanol, butanol, isopropanol and propanol; Dimethylformamide (DMF), toluene, dimethylacetamide (DMA), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, acetonitrile; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; These may be used singly or in combination, preferably methanol.

In the process for preparing a compound represented by the general formula (2) according to the present invention, the step (3) is a step of reacting the compound represented by the general formula (8) obtained in the step 2 with the compound represented by the general formula .

Specifically, the step of reacting a compound represented by the formula (8) with a halogen gas compound represented by the formula (9) in the presence of a base to obtain a halogen-substituted compound represented by the formula (10).

Examples of the base include inorganic bases such as potassium hydroxide, sodium t-butoxide, potassium t-butoxide, sodium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate and sodium hydride; Or an organic base such as N, N-diisopropylethylamine (DIPEA), 1,8-diazabicyclo [5.4.0] -7-anesene (DBU), pyridine, triethylamine, Or may be used in excess, alone or in combination, and it is preferable to use potassium hydroxide.

Also usable reaction solvents include dimethylformamide (DMF), toluene, dimethylacetamide (DMA), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, acetonitrile; Lower alcohols including isopropanol, methanol, ethanol, propanol and butanol; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; These may be used singly or in combination, and it is preferable to use dimethylformamide.

In step (4), the compound represented by formula (10) obtained in step (3) is reacted with the compound represented by formula (11) to obtain the compound represented by formula (12) .

Specifically, a step of reacting a compound represented by the formula (10) with a halogen gas compound represented by the formula (11) to obtain a halogen-substituted compound represented by the formula (12).

Examples of the reaction solvent which can be used include acetic acid, dimethylformamide (DMF), toluene, dimethylacetamide (DMA), dimethylsulfoxide (DMSO), methylene chloride, dichloroethane, water, ethyl acetate, Acetonitrile; Lower alcohols including isopropanol, methanol, ethanol, propanol and butanol; Ether solvents including tetrahydrofuran (THF), dioxane, ethyl ether, 1,2-dimethoxyethane and the like; Etc., which can be used singly or in combination, and acetic acid is preferably used.

In the method for preparing the compound represented by Chemical Formula 2 according to the present invention, in Step 5, the compound represented by Chemical Formula 12 and the compound represented by Chemical Formula 13 obtained in Step 4 are reacted to prepare a compound represented by Chemical Formula 2 .

Specifically, the step of reacting -NH of the compound represented by the formula (12) and A ³ of the compound represented by the formula (13) in the presence of a base to obtain a compound represented by the formula (2).

Examples of the base include inorganic bases such as sodium t-butoxide, potassium t-butoxide, sodium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate and sodium hydride; Or an organic base such as N, N-diisopropylethylamine (DIPEA), 1,8-diazabicyclo [5.4.0] -7-anesene (DBU), pyridine, triethylamine, Or may be used in excess, alone or in combination, and it is preferable to use sodium t-butoxide.

The starting material (the compound represented by the formula 2) Recipe 2

The compound represented by Formula 2, which is the starting material of Reaction Scheme 1,

Reacting a compound represented by the formula (14) with sodium nitrite to obtain a compound represented by the formula (15) (step 1);

Reacting a compound represented by the formula (15) obtained in the above step 1 with a compound represented by the formula (9) to obtain a compound represented by the formula (12) (step 2); And

And reacting the compound represented by the formula (12) obtained in the above step 2 with a compound represented by the formula (13) to obtain a compound represented by the formula (2): < EMI ID =

[Reaction Scheme 3]

In Scheme 3,

A < ³ > is halogen; And

R ² and n are the same as defined in the above formula (1).

In the process for preparing the compound represented by the formula (2) according to the present invention, the step (1) is a step of reacting the compound represented by the formula (14) with sodium nitrite to obtain the compound represented by the formula (15).

Specifically, the compound represented by the formula (6) is obtained by reacting -NH ₂ of the compound represented by the formula (6) and methyl and sodium nitrite to form a pentane ring.

In the method for preparing the compound represented by the formula 2 according to the present invention, the step 2 is a step of reacting the compound represented by the formula 15 obtained in the step 1 with the compound represented by the formula 9 to obtain the compound represented by the formula 12 And a specific production method can be carried out in the same manner as the production method of the step 3 to the step 3 in the above-mentioned reaction formula 2.

In the process for preparing the compound represented by the general formula (2) according to the present invention, in the step (3), the compound represented by the general formula (12) obtained in the step 2 is reacted with the compound represented by the general formula , And a specific preparation method can be carried out in the same manner as the preparation method of the above-mentioned Scheme 2 to 5. [

Specifically, the MELK-related diseases may include cancers such as breast cancer, lung cancer, bladder cancer, lymphoma, polymorphonalephritis, and cervical cancer.

As a result of evaluating the ability of the compound represented by Formula 1 according to the present invention to inhibit MELK expression, it was found that the compound of Example of the present invention inhibited MELK expression. In particular, it was found that the compounds of Examples 17 and 31 have an IC ₅₀ value of 0.4 μM or less, which suppresses the expression of MELK excellently even at a very low concentration (see Experimental Example 1).

Therefore, the compound represented by the formula (1) according to the present invention has excellent ability to inhibit MELK expression, and thus it is possible to use a PPAR agonist for the treatment of MELK-related diseases such as breast cancer, lung cancer, bladder cancer, lymphoma, Prevention or treatment.

In the pharmaceutical composition according to the present invention, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof may be administered orally or parenterally in various formulations at the time of clinical administration. In the case of formulation, For example, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like.

Examples of formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules, elixirs and troches, , Dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants (such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols). The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain binders such as starch, agar, alginic acid or sodium salts thereof Release or boiling mixture and / or absorbent, colorant, flavor, and sweetening agent.

The pharmaceutical composition containing the compound represented by Formula 1 or its pharmaceutically acceptable salt according to the present invention as an active ingredient may be administered parenterally, and parenteral administration may be carried out by subcutaneous injection, intravenous injection, intramuscular injection, It depends on the injection method.

In this case, in order to formulate the composition for parenteral administration, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof may be mixed with water or a stabilizer or a buffer to prepare a solution or a suspension, . The compositions may contain sterilized and / or preservatives, stabilizers, wettable or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, Or may be formulated according to the coating method.

The dosage of the pharmaceutical composition containing the compound of formula (I) of the present invention or its pharmaceutically acceptable salt as an active ingredient in the human body depends on the age, body weight, sex, dosage form, health condition and disease And is typically 0.1-1000 mg / day, preferably 1-500 mg / day, based on adult patients weighing 70 Kg, and may be administered at a physician or pharmacist's discretion It may be administered once to several times a day at intervals of time.

Furthermore, the pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof according to the present invention as an active ingredient can be used alone or in combination with other agents such as surgery, hormone therapy, chemotherapy And methods using biological response modifiers.

Herein, the MELK-related disease may include cancer such as breast cancer, lung cancer, bladder cancer, lymphoma, polymorphonalephritis, cervical cancer and the like.

The compound represented by the formula (1) according to the present invention acts as an inhibitor that suppresses the expression of MELK, thereby providing a health functional food composition for preventing or ameliorating MELK-related diseases such as cancer and the like, can do.

The compound represented by the formula (1) according to the present invention can be added directly to food or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement). Generally, the amount of the compound in the health food may be 0.1 to 90 parts by weight of the total food. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

In addition, the health functional beverage composition of the present invention has no particular limitation on other components other than the above-mentioned compounds as essential components in the indicated ratios, and may contain various flavoring agents or natural carbohydrates as additional components such as ordinary beverages have. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g, per 100 g of the composition of the present invention.

In addition, in addition to the above, the compound represented by the formula (1) according to the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates such as cheese, Acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compound represented by formula (1) of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

Hereinafter, examples and experimental examples of the present invention will be described in detail.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

< Manufacturing example 1 > 5- Bromo -3- Iodo -1H- Pyrazolo [3,4-b] pyridine Produce

step 1: 1H - Pyrazolo [3,4-b] pyridine -3- Amine Produce

Hydrazine hydrate (80% yield, 62 mL, 1.0 mL) was added after 2-chloronicotinonitrile (69.3 g, 0.5 mol) was dissolved in ethanol (600 mL) and refluxed for 6 hours. After completion of the reaction, the reaction was cooled to 0 ° C, and the product was washed with cold ethanol and recrystallized from ethanol to obtain 1H-pyrazolo [3,4-b] pyridin-3-amine (quantitative yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 5.59 (s, 2H), 6.95 (m, 1H), 7.19 (brs, 1H), 8.15 (m, 1H), 8.35 (m, 1H), 11.95 (s , 1H).

Steps 2 and 3: Pyrazole - < / RTI >

The 1H-pyrazolo [3,4-b] pyridin-3-amine (4.0 g, 29.82 mmol) obtained in the above step 1 was dissolved in methanol (5 mL) After cooling to 캜, diethyl ether (100 mL) and aqueous HBF ₄ solution (48% yield, 8 mL) were added. The resulting orange-colored solid was filtered and dissolved in methanol (600 mL), and then the mixed solution was treated with 0 Was added dropwise to a mixture of methanol (300 mL) and isoamyl nitrite (25 mL) in a solution of HBF ₄ (4 mL). The mixed solution was added dropwise in an amount of half over 4 hours, isoamyl nitrite (8 mL) was further added, and the other half of the mixed solution was added dropwise over 4 hours. After the end of addition, 0 RTI ID = 0.0 > 20 C < / RTI > for 1 hour, Lt; 0 > C under reduced pressure until the total volume of the solution reached 50 mL. The remaining solution was crystallized with diethyl ether (600 mL), filtered and washed with diethyl ether and water (50 mL). The solid product was treated with H ₃ PO ₂ solution (50% yield, 20 mL) and stirred at room temperature for 5 hours. The reaction was neutralized with a saturated aqueous sodium bicarbonate solution and extracted with 30% yield of isopropyl alcohol / chloroform (200 mL x 3) and purified by chromatography (ethyl acetate: n-hexane = 10% yield to 30% yield) Pyrazole-pyridine (20% yield).

¹ H NMR (CDCl ₃ , 300 MHz):? 7.21 (q, J = 4.8 Hz, 1 H), 8.18 (m, 2H), 8.68 (m,

step 4: 3 - Iodo -1H- Pyrazolo [3,4-b] pyridine Produce

The pyrazolo-pyridine (1.86 g, 15.6 mmol) obtained in steps 2 and 3 above was dissolved in DMF (dimethylformamide, 25 mL) and treated with 0 Iodine (9.9 g, 39 mmol) and KOH (3.5 g, 62.4 mmol) were placed under ice-cooling. The mixture was stirred at room temperature for 3 hours, diluted with brine (200 mL), and extracted with ethyl acetate (200 mL x 5). The organic layer was washed with Na ₂ S ₂ O ₃ solution (100 mL) and brine (100 mL), dried over magnesium sulfate and concentrated under reduced pressure to give 3-iodo-1H-pyrazolo [3,4- b] 3.75 g, 98% yield).

LC-MS: m / z 245.88.

step 5: 5 - Bromo -3- Iodo -1H- Pyrazolo [3,4-b] pyridine Produce

3-Iodo-1H-pyrazolo [3,4-b] pyridine (3.75 g, 98% yield), acetic acid (40 mL) and bromine (1.6 mL, 30.6 mmol) obtained in step 4 were placed in a sealed tube . The reaction mixture was diluted with 100 Lt; 0 > C for 12 hours, cooled to room temperature, and then neutralized with sodium hydrogencarbonate solution. After the organic layer was extracted with ethyl acetate and washed with Na ₂ S ₂ O ₃ solution, concentrated, purified by chromatography (ethyl acetate: dichloromethane = 5% yield) to give 5-bromo-3-iodo -1H- pyrazol to 3,4-b] pyridine (42% yield).

^{_{1 H NMR (DMSO-d 6}} , 300HMz): δ 8.41 (d, J = 1.8 Hz, 1H), 8.68 (d, J = 2.1 Hz, 1H); LC-MS: m / z 277.9, 279.8.

< Manufacturing example 2> 5- Bromo -3- Iodo -1H- Pyrazolo [3,4-c] pyridine Produce

Step 1: Preparation of 2-bromo-4-methyl-5-nitropyridine

4-Methyl-5-nitropyridin-2-ol (4.0 g, 26 mmol) was dissolved in 1,2-dichloroethane and then phosphorus oxybromide (POBr ₃ ) C < / RTI > for 24 hours. After cooling, the solvent was concentrated under reduced pressure, and the residue was dissolved in 30% 2-propanol / chloroform and washed with water. The organic layer was purified by chromatography (5% ethyl acetate / hexane) to give 2-bromo-4-methyl-5-nitropyridine (4.8 g, 86% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 2.64 (s, 3H), 7.53 (s, 1H), 8.96 (s, 1H).

step 2: 6 - Bromo -4- Methyl pyridine - Amine Produce

2-Bromo-4-methyl-5-nitropyridine (4.8 g, 22 mmol) obtained in the above step 1 was dissolved in THF (tetrahydrofuran, 120 mL), Raney Ni (Raney nickel, 2.4 g) And the mixture was stirred at room temperature under hydrogen (balloon) for 24 hours. The reaction was filtered through celite, concentrated and purified by chromatography (50-60% ethyl acetate / hexanes) to give 6-bromo-4-methylpyridine-amine (4g, 96% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 2.13 (s, 3H), 3.68 (s, 2H), 7.12 (s, 1H), 7.76 (s, 1H).

step 3: 5 - Bromo - 1H- Pyrazolo [3,4-c] pyridine Produce

(4.0 g, 21.4 mmol) obtained in the above Step 2 was dissolved in acetic acid (300 mL) and then sodium nitrite (1.47 g, 21.4 mmol) in water (2.5 mL) Was added. After stirring at room temperature for 3 days, the solvent was removed under reduced pressure and the residue was extracted with 30% 2-propanol / chloroform (200 mL). The organic layer was washed with aqueous sodium hydrogen carbonate solution and then chromatographed : n-hexane = 10% to 20%) to obtain 5-bromo-1H-pyrazolo [3,4-c] pyridine (3.3 g, 78% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 7.99 (s, 3H), 8.75 (s, 1H), 8.80 (s, 1H).

step 4: 5 - Bromo -3- Iodo -1H- Pyrazolo [3,4-c] pyridine Produce

5-bromo-1H-pyrazolo [3,4-c] pyridine (3.3 g, 16.6 mmol) obtained in the above step 3 was dissolved in DMF (300 mL) and NIS (N-iodosuccinimide, 5.6 g, 25 mmol). The reaction was stirred at room temperature for 12 hours and the reaction was diluted with 30% 2-propanol / chloroform (200 mL), washed with saturated aqueous Na ₂ S ₂ O ₃ (200 mL) and chromatographed (ethyl acetate: n -Hexane = 15%) to obtain 5-bromo-3-iodo-1H-pyrazolo [3,4-c] pyridine (5.2 g, 96% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 7.66 (s, 1H), 8.77 (s, 1H).

< Example 1 > 1- (2- Chlorobenzyl ) -N- (3- Chlorophenyl ) -5- (pyridin-3-yl) -1H- Pyrazole [3,4-b] pyridin-3-amine

step 1: 5 - Bromo -1- (2- Chlorobenzyl ) -3- Iodo -1H- Pyrazolo [3,4-b] pyridine Produce

To a solution of 5-bromo-3-iodo-1H-pyrazolo [3,4-b] pyridine (1.26 g, 3.9 mmol) obtained in Preparation Example 1 in 20 mL of DMF T-BuONa (0.45 g, 4.6 mmol) under ice-cooling, and the mixture was stirred for 15 minutes. 2-Chlorobenzyl bromide (0.6 mL, 4.6 mmol) was added, and the mixture was stirred at room temperature for 24 hours. The organic layer was concentrated and purified by chromatography (n-hexane: dichloromethane = 1: 1) to obtain 5-bromo-1- (2-chloro- Benzyl) -3-iodo-1H-pyrazolo [3,4-b] pyridine (92% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 5.78 (s, 2H), 6.92 (d, J = 7.5 Hz, 1H), 7.19 (m, 2H), 7.38 (d, J = 7.5 Hz, 1H), 8.10 (d, J = 1.8 Hz, 1 H), 8.59 (d, J = 2.1 Hz, 1 H).

Step 2: Preparation of 5-bromo-1- (2-chlorobenzyl) -N- (3-chlorophenyl) -lH-pyrazolo [3,4- b] pyridin-

(2-chlorobenzyl) -3-iodo-1H-pyrazolo [3,4-b] pyridine (0.1 g, 0.22 mmol) obtained in the above step 1 was dissolved in 5 mL of a microwave vial. Was added to toluene (4 mL), treated with nitrogen for 15 min, and then added to a solution of 3-chloroaniline (23 μL, 0.22 mmol), Cs ₂ CO ₃ (0.64 g, 2.2 mmol) and xantphos The gas was removed. Pd ₂ dba ₃ (tris (dibenzylideneacetone) dipalladium, 10 mg, 5 mol%) was added, and 130 Lt; 0 > C for 60 minutes. After completion of the reaction, the reaction product was filtered through celite, and the filtrate was concentrated and purified by chromatography (ethyl acetate: n-hexane) to obtain 5-bromo- 1- (2- chlorobenzyl) -N- ) - lH-pyrazolo [3,4-b] pyridin-3-amine (26% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 5.71 (s, 2H), 6.30 (s, 1H), 6.92 (d, J = 7.5 Hz, 1H), 6.98 (d, J = 7.5 Hz, 1H), 7.21 (m, 4H), 7.39 (m, 2H), 8.04 (s, IH), 8.54 (s, IH).

step 3: 1 -(2- Chlorobenzyl ) -N- (3- Chlorophenyl ) -5- (pyridin-3-yl) -1H- Pyrazole [3,4-b] pyridin-3-amine

(2-chlorobenzyl) -N- (3-chlorophenyl) -1H-pyrazolo [3,4-b] pyridine-3-amine 60 mg, 0.14 mmol), 3-pyridine boronic acid (22 mg, 0.18 mmol) and K ₂ CO ₃ (2 M, 0.7 mL, 1.4 mmol) were added to DMA (dimethylacetamide, 5 mL) Degassed. Pd (dppf) Cl ₂ ([1,1'-bis (diphenylphosphino) ferrocene] palladium (II) dichloride, 10 mg, 10 mol% Lt; 0 > C for 30 minutes. After completion of the reaction, the reaction product was filtered through celite, and the filtrate was washed with brine (10 mL x 5), concentrated and purified by chromatography (ethyl acetate: DCM (dichloromethane) &Lt; / RTI >

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 5.79 (s, 2H), 6.62 (s, 1H), 6.93 (m, 1H), 7.03 (d, J = 7.5 Hz, 1H), 7.15-7.25 (m, 1H), 7.41 (m, 2H), 7.50 (s, 1H), 7.89 (d, J = 7.8 Hz, 1H) 8.86 (s, 1 H).

Example 2-16 shown in the following Table 1 was prepared by a similar method to that of Example 1 above.

Example constitutional formula Compound name ¹ H NMR 2

Preparation of l- (2-chlorobenzyl) -N-phenyl-5- (pyridin-3-yl) -lH- pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 5.78 (s, 2H), 6.61 (s, 1H), 6.97 (m, 2H), 7.12-7.30 (m, 3H), 7.33 (m, 2H), 7.41 ( J = 1.8 Hz, 1H), 8.63 (d, J = 3.9 Hz, 1H), 8.73 (d, J = 2.1 Hz, 1H), 8.85 (s, 1 H) 3

3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ5.72 (s, 2H), 6.93 (m, 1H), 6.99 (m, 1H), 7.20-7.33 (m, 3H), 7.38-7.46 (m, 2H) , 7.88 (m, 1 H), 8.12 (m, 1 H), 8.54 (m, 4

L- (2-chlorobenzyl) -N- (2,3-dichlorophenyl) -5- (pyridin-3- yl) -lH- pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ5.81 (s, 2H), 6.97-7.05 (m, 3H), 7.11-7.23 (m, 3H), 7.43 (m, 2H), 7.91 (d, J = 7.8 Hz, 2H), 8.11 (d, J = 2.1 Hz, 1H), 8.66 (d, J = 3.9 Hz, 1H) , 1H) 5

Pyrazolo [3,4-b] pyridin-3-ylamine (Compound 1) was obtained in the same manner as in 1- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ5.79 (s, 2H), 6.64 (t, J = 8.4 Hz, 1H), 6.99 (s, 1H), 6.99 (d, J = 7.2 Hz, 1H), (M, 2H), 7.89 (d, J = 8.1 Hz, 1H), 8.13 (d, J = 7.6 Hz, 1H), 7.13-7.23 , 8.75 (d, J = 3.6 Hz, 1H), 8.75 (d, J = 1.8 Hz, 1H) 6

Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 2- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ5.77 (s, 2H), 6.41 (s, 1H), 7.01 (m, 3H), 7.13-7.22 (m, 2H), 7.35-7.43 (m, 4H) , 7.87 (m, 1 H), 8.04 (d, J = 1.8 Hz, 1 H), 8.63 7

Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 1- (2-chlorobenzyl) ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ2.32 (s, 3H), 5.80 (s, 2H), 6.20 (s, 1H), 6.60 (m, 1H), 7.02 (d, J = 7.2 Hz, 1H J = 7.8 Hz, 1H), 8.02 (s, 1H), 8.64 (d, J = 4.8 Hz, 1 H), 8.77 (d, J = 2.1 Hz, 1 H), 8.84 8

L- (2-chlorobenzyl) -N- (2,5-dimethylphenyl) -5- (pyridin- 3- yl) -lH- pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ2.26 (s, 3H), 2.31 (s, 3H), 5.78 (s, 2H), 6.12 (s, 1H), 6.77 (d, J = 7.2 Hz, 1H 2H), 7.40 (m, 2H), 7.85 (d, J = 7.5 Hz, J = 7.8 Hz, 1H), 7.93 (d, J = 1.5 Hz, H), 8.62 ) 9

Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 2- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.74 (s, 6H), 5.77 (s, 2H), 6.10 (s, 1H), 6.54 (s, 1H), 6.60 (d, J = 2.1 Hz, 2H ), 7.40 (d, J = 7.5 Hz, 1H), 7.12-7.22 (m, 2H), 7.41 (m, 2H), 7.89 (td, J = 1.5, 6.3 Hz, 1H) 2.1 Hz, 1H), 8.64 (d, J = 3.6 Hz, 1H), 8.75 10

Pyrazolo [3,4-b] pyridin-3-amine (compound 1) was prepared in accordance with the general method of example 1 from 2- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.52 (s, 3H), 5.78 (s, 2H), 6.88 (d, J = 6.9 Hz, 1H), 7.12 (d, J = 8.7 Hz, 2H), (M, 2H), 7.37-7.44 (m, 3H), 7.71 (d, J = 7.5 Hz, 1H), 8.59 (d, J = 4.8 Hz, 1H), 8.67 , 2H) 11

3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ5.83 (s, 2H), 6.87 (t, J = 4.8 Hz, 1H), 7.01 (d, J = 7.8 Hz, 1H), 7.23-7.35 (m, 2H ), 7.51 (dd, J = 0.9,6.9 Hz, 1H), 7.56 (q, J = 4.8 Hz, 1H), 8.27-8.30 (d, J = 1.5,3.3 Hz, 1H), 8.66 (d, J = 2.1 Hz, 1H), 8.96 (d, J = 2.1 Hz, 1H), 9.13 (s, 1 H) 12

L- (3-methoxybenzyl) -N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.77 (s, 3H), 5.62 (s, 2H), 6.42 (s, 1H), 6.81 (d, J = 8.4 Hz, 1H), 6.94-6.98 (m J = 2.1 Hz, 1H), 8.63 (d, J = 3.0 Hz, 1H), 8.73 (d, J = (d, J = 2.1Hz, 1 H), 8.83 (s, 1 H) 13

Pyrazolo [3,4-b] pyridin-3-amine hydrochloride (prepared according to the method described in Example 1) ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.77 (s, 3H), 5.64 (s, 2H), 6.82 (dd, J = 2.1, 6.0 Hz, 1H), 6.85-6.99 (m, 4H), 7.18 (M, 2H), 7.76-7.93 (m, 2H), 8.06 (d, J = 1.8 Hz, 1H), 8.64 8.76 (d, J = 1.5 Hz, 1 H) 14

Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 3- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.77 (s, 3H), 5.62 (s, 2H), 6.79 (dd, J = 1.8, 6.6 Hz, 1H), 6.89-7.00 (m, 4H), 7.15 1H), 7.85 (d, J = 7.8 Hz, 1H), 8.05 (d, J = 1.8 Hz, 1H), 8.83 (s, 1H), 8.62 (d, J = 4.5 Hz, 15

Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 3- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ3.77 (s, 3H), 5.62 (s, 2H), 6.16-6.69 (m, 2H), 6.81 (d, J = 7.8 Hz, 1H), 6.97-7.04 (d, J = 3.0, 4.8 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 8.07 (m, 2H) (d, J = 1.8 Hz, 1H), 8.63 (d, J = 4.8 Hz, 1H) 16 Pyrazolo [3,4-b] pyridin-3-amine (Compound A) was prepared in accordance with the general method of example 1 from 5- ^{_{1 H NMR (CDCl 3, 300HMz}} ): δ2.31 (s, 3H), 3.78 (s, 3H), 5.63 (s, 2H), 6.15 (s, 1H), 6.59 (td, J = 1.8, 6.3 Hz J = 8.1 Hz, 1H), 7.30 (m, 1H), 7.34 (dd, 1H), 6.82 (dd, J = 1.8,8.1 Hz, J = 2.1, 9.2 Hz, 1H), 7.42 (dd, J = 2.7, 4.5 Hz, 1H), 7.88 1H), 8.76 (d, J = 1.5 Hz, 1H), 8.84

< Example 17> N- (3- Chlorophenyl ) -5- (1- (Piperidin-4-yl) -1H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

step 1: 5 - Bromo -3- Iodo -1- (4-methoxybenzyl) -lH- Pyrazolo [3,4-b] pyridine Produce

To a DMF (50 mL) solution of 5-bromo-3-iodo-1H-pyrazolo [3,4-b] pyridine (1.79 g, 5.5 mmol) obtained in the above- T-BuONa (0.64 g, 6.6 mmol) under ice-cooling, and the mixture was stirred for 15 minutes. 4-Methoxybenzyl bromide (0.96 mL, 6.6 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate (200 mL), washed with brine (200 mL x 5), and the organic layer was concentrated and purified by chromatography (10% ethyl acetate: n-hexane) to obtain 5-bromo-3- - (4-methoxybenzyl) -lH-pyrazolo [3,4-b] pyridine (quantitative yield).

Step 2: Preparation of 5-bromo-1- (4-methoxybenzyl) -N- (3-chlorophenyl) -1H-pyrazolo [3,4 b] pyridin-

3-iodo-1- (4-methoxybenzyl) -1H-pyrazolo [3,4-b] pyridine (0.3 g, 0.67 mmol) obtained in the above step 1, BINAP -Bis (diphenylphosphino) -1,1'-binaphthyl, 30 mol%) was used in place of 5-bromo-1- ( (4-methoxybenzyl) -N- (3-chlorophenyl) -lH-pyrazolo [3,4-b] pyridine-3-amine (26% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 3.76 (s, 3H), 5.51 (s, 2H), 6.21 (s, 1H), 6.83 (d, J = 8.4 Hz, 2H), 6.91 (d, J = J = 8.1 Hz, 1H), 7.33 (m, 3H), 7.98 (d, J = 2.1 Hz, 1H), 8.53 (d, J = 2.1 Hz, 1H).

Step 3: tert -Butyl 4- (4- (1- (4-methoxybenzyl) -3- (3-chlorophenylamino) -1H-pyrazolo [3,4- b] -Pyrazol-1-yl) piperidine-1-carboxylate

Pyrazolo [3,4b] pyridin-3-amine (75 mg, 0.17 mmol) obtained in the above Step 2, (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-pyrazol-1 -yl) piperidine- 1-carboxylate (0.22 mmol) was used in place of tert-butyl 4- (4- (1- (4-methoxybenzyl) -3- Pyrazolo [3,4-b] pyridin-5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (69% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 1.48 (s, 9H), 1.92 (m, 2H), 2.13 (m, 2H), 2.89 (m, 2H), 3.73 (s, 3H), 4.26 (m, (M, 2H), 7.35 (d, J = 7.2 Hz, 2H), 5.54 (s, 2H), 6.53 (d, J = 7.8 Hz, 2H), 7.40 (s, 1H), 7.61 (s, 1H), 7.75 (s, 1H), 7.88

Step 4: N- (3- Chlorophenyl ) -5- (1- (Piperidin-4-yl) -1H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

(4-methoxybenzyl) -3- (3-chlorophenylamino) -lH-pyrazolo [3,4-b] pyridine 5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (1 mmol) was added trifluoroacetic acid (2 mL) and stirred at 75 ° C for 24 hours. After completion of the reaction, the reaction product was cooled and the solvent was concentrated under reduced pressure. The filtrate was added to a saturated aqueous solution of sodium hydrogencarbonate (10 mL), stirred for 30 minutes, extracted with 30% 2-propanol / acetonitrile solution, concentrated and purified by prep-HPLC (eluent: water / acetonitrile) The objective compound (36% yield) was obtained.

¹ H NMR (CD ₃ OD, 300 MHz):? 2.30 (m, 4H), 3.24 (m, 1H), 3.58 ), 7.22 (t, J = 8.4 Hz, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.80 , &Lt; / RTI > 1H), 8.69 (s, 1H).

< Example 18 > N- (3- Chlorophenyl ) -5- (1H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

Step 1: N- (3- Chlorophenyl ) -1- (4-methoxybenzyl) -5- (1 H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

(3-chlorophenyl) -1H-pyrazolo [3,4b] pyridin-3-amine (75 mg , 0.17 mmol) and (1H-pyrazol-4-yl) boronic acid (0.22 mmol) were used in the same manner as in step 3 of Example 17 to obtain N- (3-chlorophenyl) Pyrazolo [3,4-b] pyridin-3-amine (47% yield) was obtained as a pale-yellow amorphous solid.

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 3.73 (s, 3H), 5.49 (s, 2H), 6.84 (m, 3H), 7.21 (m, 1H), 7.29 (m, 2H), 7.45 (m, 1H), 7.80 (s, 1H), 8.01 (s, 2H), 8.40 (s, 1H), 8.74

Step 2: N- (3- Chlorophenyl ) -5- (1H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

(1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridine- (3-chlorophenyl) -5- (lH-pyrazol-4-yl) -1H-1-indolecarboxamide was prepared by the same procedure as in the step 4 of Example 17, Pyrazolo [3,4-b] pyridin-3-amine (35% yield).

^{_{1 H NMR (CD 3 OD,}} 300HMz): δ 6.85 (m, 1H), 7.22 (m, 1H), 7.44 (m, 1H), 7.83 (s, 1H), 8.05 (m, 2H), 8.49 (s , &Lt; / RTI > 1H), 8.72 (s, 1H).

< Example 19> N- (3- Chlorophenyl ) -5- (1- methyl -1H- Pyrazole -4-yl) -1H- Pyrazolo [3,4-b] pyridine -3-amine

Step 1: N- (3- Chlorophenyl ) -1- (4-methoxybenzyl) -5- (1- methyl -1H- Pyrazole -4-yl) -1H-pyrazolo [3,4-c] pyridin-3-amine

(3-chlorophenyl) -1H-pyrazolo [3,4b] pyridin-3-amine (75 mg , 0.17 mmol) and (1-methyl-1H-pyrazol-4-yl) boronic acid (0.22 mmol) were used in the same manner as in step 3 of Example 17, LH-pyrazolo [3,4-c] pyridine-3-amine (76%) was obtained as colorless crystals from 4- (4-methoxybenzyl) Yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 3.73 (s, 3H), 3.91 (s, 3H), 5.53 (s, 2H), 6.83 (m, 3H), 6.87 (m, 1H), 7.16 (d, 1H, J = 5.1 Hz, 2H), 7.34 (d, J = 8.7 Hz, 2H), 7.43 (s, 1H), < / RTI > 8.64 (d, J = 2.1 Hz, 1H).

Step 2: Preparation of N- (3-chlorophenyl) -5- (1 -methyl-1 H-pyrazol-4-yl) -lH- pyrazolo [3,4- b] pyridin-

(3-chlorophenyl) -1- (4-methoxybenzyl) -5- (1 -methyl-1 H- pyrazol-4-yl) -1H- pyrazolo [3,4- c] pyridin-3-amine (1 mmol) obtained in Step 4 of Example 17, the target compound (36% yield) was obtained.

^{_{1 H NMR (CD 3 OD,}} 300HMz): δ 3.95 (s, 3H), 6.84 (d, J = 7.8 Hz, 1H), 7.21 (t, J = 8.1Hz, 1H), 7.44 (d, J = 8.1 1H), 7.81 (s, 1H), 7.85 (s, 1H), 8.0 (s, 1H), 8.48

< Example 20> N- (3,4- Dichlorophenyl ) -5- (1- methyl -1H- Pyrazole -4-yl) -1H- Pyrazole [3,4-b] pyridin-3-amine

Step 1: N- (3,4-Dichlorophenyl) -l- (4-methoxy-benzyl) -5- (1 -methyl-lH- pyrazol- 4-b] pyridine-3-amine

(3-chlorophenyl) -1H-pyrazolo [3,4b] pyridin-3-amine (75 mg , (0.17 mmol) and (1-methyl-1H-pyrazol-4-yl) boronic acid (0.22 mmol) were used in the same manner as in step 3 of Example 17, LH-pyrazolo [3,4-b] pyridin-3-ylmethyl) -1H-pyrazol- Amine (76% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 3.75 (s, 3H), 3.95 (s, 3H), 5.54 (s, 2H), 6.58 (s, 1H), 6.83 (d, J = 8.7 Hz, 2H) 2H), 7.35 (d, J = 8.7 Hz, 2H), 7.56 (d, J = 2.4 Hz, 1H), 7.59 , 7.73 (s, 1H), 7.87 (d, J = 1.8 Hz, 1H), 8.66 (d, J = 2.1 Hz, 1H).

Step 2: Preparation of N- (3,4-dichlorophenyl) -5- (l -methyl-lH-pyrazol-4-yl) -lH- pyrazolo [3,4- b] pyridin-

(4-methoxybenzyl) -5- (1-methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3 , 4-b] pyridine-3-amine (1 mmol) was used in place of 4-chloropyridine.

^{_{1 H NMR (DMSO-d 6}} , 300HMz): δ 3.94 (s, 3H), 7.42 (d, J = 5.7 Hz, 1H), 7.58 (d, J = 6.3 Hz, 1H), 7.81 (s, 1H) , 8.04 (s, IH), 8.20 (s, IH), 8.45 (s, IH), 8.75

< Example 21> N- (3,5- Dichlorophenyl ) -5- (1- methyl -1H- Pyrazole -4-yl) -1H- Pyrazole [3,4-b] pyridin-3-amine

Step 1: N- (3,5-Dichlorophenyl) -l- (4-methoxy-benzyl) -5- (1 -methyl-lH- pyrazol- 4-b] pyridine-3-amine

To a solution of 5-bromo-1- (4-methoxybenzyl) -N- (3,5-dichlorophenyl) -1 H-pyrazolo [3,4b] pyridin- (75 mg, 0.17 mmol) and (1-methyl-1H-pyrazol-4-yl) boronic acid (0.22 mmol) (L, 4-dimethoxy-benzyl) -5- (1 -methyl- lH- pyrazol-4-yl) -lH- pyrazolo [3,4- b] -3-amine (50% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 3.75 (s, 3H), 3.94 (s, 3H), 5.54 (s, 2H), 6.85 (m, 3H), 6.98 (s, 1H), 7.29 (d, J = 1.5 Hz, 2H), 7.37 (d, J = 8.7 Hz, 2H), 7.58 (s, J = 1.8 Hz, 1H).

Step 2: Preparation of N- (3,5-dichlorophenyl) -5- (l -methyl-lH-pyrazol-4-yl) -lH-pyrazolo [3,4- b] pyridin-

To a solution of N- (3,5-dichlorophenyl) -1- (4-methoxybenzyl) -5- (1 -methyl-1 H-pyrazol-4-yl) -1H- pyrazolo [3 , 4-b] pyridine-3-amine (1 mmol) was used in place of 4-chloropyridine.

¹ H NMR (CD ₃ OD, 300 MHz):? 3.96 (s, 3H), 6.87 (s, IH), 7.66 (s, 2H), 7.87 , &Lt; / RTI > 1H), 8.69 (s, 1H).

< Example 22> N- (3,5- Dichlorophenyl ) -5- (1- (Piperidin-4-yl) -1H- Pyrazole Yl) -lH-pyrazolo [3,4-b] pyridin-3-amine

Step 1: tert -Butyl 4- (4- (3 - ((3,5-dichlorophenyl) amino) -1- (4- methoxybenzyl) -lH- pyrazolo [3,4- b] 5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

To a solution of 5-bromo-1- (4-methoxybenzyl) -N- (3,5-dichlorophenyl) -1 H-pyrazolo [3,4b] pyridin- (75 mg, 0.17 mmol), tert-butyl 4- (4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan- Butyl) -4- (4- (3 - ((3, < / RTI > LH-pyrazol-l-yl) piperidine-l- (4-methoxybenzyl) -lH-pyrazolo [3,4- 1-carboxylate (50% yield).

^{_{1 H NMR (CDCl 3, 300HMz}} ): δ 1.49 (s, 9H), 1.90 (m, 2H), 2.11 (m, 2H), 2.88 (m, 2H), 3.73 (s, 3H), 4.28 (m, 1H), 7.73 (s, 1H), 7.73 (s, 2H), 6.87 (s, ), 7.92 (d, J = 2.1 Hz, 1H), 8.65 (d, J = 1.8 Hz, 1H).

Step 2: N- (3,5-Dichlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol-4-yl) -lH- pyrazolo [3,4- b ] Pyridine-3-amine

3,4-b] thiophene obtained in the above step 1 was reacted with tert-butyl 4- (4- (3 - ((3,5- dichlorophenyl) amino) 1-carboxylate (1 mmol) was used in place of 4-chloro-pyridin-5-yl) 10% yield).

^{_{1 H NMR (CD 3 OD,}} 300HMz): δ 2.37 (m, 4H), 3.24 (m, 1H), 3.57 (m, 3H), 4.62 (m, 1H), 6.87 (s, 1H), 7.45 (s (S, 1H), 7.66 (s, 1H), 7.95 (s, 1H).

Examples 23 to 29 shown in the following Table 2 were produced by the same production method as in Example 17. [

Example constitutional formula Compound name ¹ H NMR 23

N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ6.79 (t, J = 7.2 Hz, 1H), 7.18 (t, J = 7.5 Hz, 2H), 7.53 (d, J = 7.5 Hz, 2H), 7.88 (d, J = 3.3,5.7 Hz, 1H), 8.60 (m, 2H), 8.65 (d, J = 5.4 Hz, 1H) 24

N- (2-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ6.90 (t, J = 7.5 Hz, 1H), 7.22 (t, J = 7.5 Hz, 1H), 7.39 (d, J = 7.8 Hz, 1H), 8.03 (d, J = 5.4 Hz, 1 H), 8.08 (t, J = 7.8 Hz, 1 H), 8.69 (m, 1 H), 9.21 (s, 1 H) 25

N- (3-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ6.83 (dt, J = 0.9, 6.3 Hz, 1H), 7.20 (t, J = 8.4 Hz, 1H), 7.41 (dt, J = 0.9, 6.3 Hz, J = 1.5 Hz, 1H), 7.79 (t, J = 0.9 Hz, 1H), 8.04 1H), 9.12 (s, 1 H) 26

(3-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ6.49 (m, 1H), 7.11-7.22 (m, 2H), 7.52 (td, J = 2.1, 7.8 Hz, 1H), 7.86 (dd, J = 2.7 , 5.4 Hz, 1H), 8.54 (m, 1H), 8.58 (d, J = 2.4 Hz, 1H), 8.65 9.0 (s, 1 H) 27

(4-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ7.0 (t, J = 8.7 Hz, 2H), 7.63 (q, J = 4.5 Hz, 2H), 8.06 (t, J = 7.2 Hz, 1H), 8.68 (d, J = 1.8 Hz, 1 H), 8.79 (m, 3H), 9.15 28

(5-fluoro-2-methylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ2.35 (s, 3H), 6.57 (t, J = 2.4, 4.2 Hz, 1H), 7.13 (t, J = 7.5 Hz, 1H), 7.59 (dt, J = 2.1, 7.2 Hz, 1H), 8.06 (td, J = 1.8, 6.0 Hz, 1H), 8.69 Hz, 1 H), 9.18 (s, 1 H) 29

N- (2,5-dimethylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- b] pyridin- ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ2.24 (s, 3H), 2.29 (s, 3H), 6.78 (d, J = 6.2 Hz, 1H), 7.05 (d, J = 6.2 Hz, 1H) , 7.41 (s, IH), 8.10 (t, J = 7.2 Hz, IH), 8.58 (s, IH), 8.81 (m, 3H), 9.19

< Example 30> N- (3- Chlorophenyl ) -5- (pyridin-3-yl) -1H- Pyrazolo [3,4-c] pyridine -3-amine

Step 1: Preparation of 5-bromo-3-iodo-1- (4-methoxybenzyl) -1H-pyrazolo [3,4-c] pyridine

Was obtained in the same manner as in Example 17, except that 5-bromo-3-iodo-1H-pyrazolo [3,4-c] pyridine (2.0 g, 6.2 mmol) 1, to give 5-bromo-3-iodo-1- (4-methoxybenzyl) -1H-pyrazolo [3,4- c] pyridine (2.2 g, 80% yield) .

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 3.78 (s, 3H), 5.59 (s, 2H), 6.85 (d, J = 8.7 Hz, 2H), 7.23 (d, J = 8.7 Hz, 2H), 7.57 (s, 1 H), 8.48 (s, 1 H).

step 2: 5 - Bromo -N- (3- Chlorophenyl ) -1- (4-methoxybenzyl) -1H- Pyrazolo [3,4-c] pyridine -3-amine

Pyrazolo [3,4-c] pyridine (0.3 g, 0.67 mmol), Pd (dppf) Cl The procedure of Example 17 was repeated except for using ₂ (50 mg, 10 mol%), dppf (1,1'-bis (diphenylphosphino) ferrocene, 74 mg, 20 mol% (3-chlorophenyl) -l- (4-methoxybenzyl) -lH-pyrazolo [3,4-c] pyridine-3-amine (0.12 g, 40% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 3.77 (s, 3H), 5.45 (s, 2H), 6.28 (s, 1H), 6.85 (d, J = 8.7 Hz, 2H), 6.93 (d, J 2H), 7.21 (m, 2H), 7.25 (d, J = 8.7Hz, 2H), 7.45 (s, 1H), 7.63 (s, 1H), 8.48 (s, 1H).

Step 3: N- (3- Chlorophenyl ) -1- (4-methoxybenzyl) -5- (pyridin-3-yl) -1H- Pyrazolo [3,4-c] pyridine -3-amine

Pyrazolo- [3,4-c] pyridin-3-amine (51 mg, 0.25 mmol) obtained in the above Step 2, 0.17 mmol) and 3-pyridinoboronic acid (18 mg, 0.15 mmol) were used in place of N- (3-chlorophenyl) -1- Pyrazolo [3,4-c] pyridin-3-amine (37 mg, 74% yield) was obtained as colorless crystals.

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 3.78 (s, 3H), 5.52 (s, 2H), 6.72 (s, 1H), 6.87 (d, J = 8.4 Hz, 2H), 6.94 (m, 1H ), 7.28 (m, 4H), 7.38 (s, IH), 7.55 (s, IH), 7.89 (s, IH), 8.28 (d, J = 7.2 Hz, , &Lt; / RTI > 1H), 8.83 (s, 1H), 9.17 (s, 1H).

Step 4: Preparation of N- (3-chlorophenyl) -5- (pyridin-3-yl) -lH- pyrazolo [3,4- c] pyridin-

(3-chlorophenyl) -1- (4-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4- c] pyridin- (40% yield) was obtained by carrying out the same method as in the step 4 of Example 17, except that the title compound (35 mg, 0.08 mmol) was used.

^{_{1 H NMR (CD 3 OD,}} 300HMz): δ 6.85 (d, J = 7.2 Hz, 1H), 7.23 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.8 Hz, 1H), 7.46 ( 1H, J = 7.5 Hz, 1H), 7.81 (s, 1H), 8.10 (t, J = 6.9 Hz, 1H), 8.58 9.11 (d, J = 7.8 Hz, 1 H), 9.44 (s, 1 H).

< Example 31> N- (3- Chlorophenyl ) -5- (1- (Piperidin-4-yl) -1H- Pyrazole -4-yl) -1H- blood Razolo [3,4-c] pyridine-3-amine

Step 1: tert-Butyl 4- (4- (3 - ((3-chlorophenyl) amino) -1- (4- methoxybenzyl) -lH- pyrazolo [3,4- c] pyridin- ) -1H-pyrazol-1-yl) piperidine-1-carboxylate

[0353] To a solution of 5-bromo-N- (3-chlorophenyl) -l- (4-methoxy- benzyl) -lH- pyrazolo- [3,4- c] Amine (51 mg, 0.17 mmol), tert-butyl 4- (4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan- Butyl 4- (4- (3 - ((3 (R) -pyrrolidin-1 -yl) -piperidine-1-carboxylate was obtained in the same manner as in step 3 of Example 17, LH-pyrazol-l-yl) piperidine-l- (4-methoxybenzyl) -lH- pyrazolo [3,4- c] pyridin- Carboxylate (41% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 1.84 (s, 9H), 1.85 (m, 2H), 2.07 (m, 2H), 2.85 (m, 2H), 3.76 (s, 3H), 4.23 (m 1H), 7.20 (m, 1H), 7.28 (d, J = 8.2 Hz, (m, 3H), 7.57 (m, IH), 7.62 (s, IH), 7.83 (s, IH), 7.88 (s, IH), 8.68

Step 2: N- (3-Chlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol- Preparation of 3-amine

3,4-c] pyridin-5-ylamine obtained in the above step 1, and tert -butyl 4- (4- (3- ((3- chlorophenyl) amino) -Yl) -lH-pyrazol-1-yl) piperidine-1-carboxylate was used in place of 4-chlorobenzyl chloride in step 4 above to give the title compound (35% yield).

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 2.38 (m, 5H), 3.62 (m, 3H), 4.65 (m, 1H), 6.90 (d, J = 8.2 Hz, 1H), 7.30 (t, J (D, J = 7.8 Hz, 1H), 7.50 (d, J = 7.8 Hz, 1H), 7.85 (s, 1H), 8.10 , 1H).

< Example 32> N- (3,4- Dichlorophenyl ) -5- (1- (Piperidin-4-yl) -1H- Pyrazole -4-yl) -1H-pyrazolo [3,4-c] pyridin-3-amine

Step 1: tert -Butyl 4- (4- (3 - ((3-chlorophenyl) amino) -1- (4- methoxybenzyl) -lH- pyrazolo- [3,4- c] Yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

3,4-c] pyridin-3-amine (51 mg, 0.17 mmol), and 5-bromo-N- (3,4- dichlorophenyl) (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -lH-pyrazol-l-yl) -piperidin- Carboxylate (0.15 mmol) was used in place of tert-butyl 4- (4- (3 - ((3-chlorophenyl) amino) -1- (41% yield) was obtained as a pale yellow solid from < RTI ID = 0.0 > .

^{_{1 H NMR (CDCl 3, 300}} HMz): δ 1.49 (s, 9H), 1.85 (m, 2H), 2.10 (m, 2H), 2.85 (m, 2H), 3.76 (s, 3H), 4.23 (m 1H), 7.72 (s, 1H), 7.64 (s, 2H), 6.83 (d, J = 8.4 Hz, 2H) , 7.82 (s, 1 H), 7.88 (s, 1 H), 8.68 (s, 1 H).

Step 2: N- (3,4-Dichlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol-4-yl) -lH- pyrazolo [3,4- c ] Pyridine-3-amine

[0157] To a solution of tert-butyl 4- (4- (3 - ((3-chlorophenyl) amino) -1- (4- methoxybenzyl) -1 H- pyrazolo- [3,4- c] (20% yield) was obtained by carrying out the same method as in the step 4 of Example 17, except using 5-ethoxy-5- (4-fluorophenyl) .

^{_{1 H NMR (CD 3 OD,}} 300 HMz): δ 2.36 (m, 5H), 3.62 (m, 3H), 4.68 (m, 1H), 7.39 (m, 1H), 7.49 (m, 1H), 8.00 ( (s, 1H), 8.06 (s, 1H), 8.22 (d, J = 5.4 Hz, 2H), 8.90

Examples 33-35 shown in the following Table 3 were prepared by the same method as in Example 30.

Example constitutional formula Compound name ¹ H NMR 33

LH-pyrazolo [3,4-c] pyridine-1-carboxylic acid ethyl ester was prepared from N- (3,4-dichlorophenyl) 3-amine ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ 2.37 (m, 5H), 3.62 (m, 3H), 4.77 (m, 1H), 6.89 (s, 1H), 7.66 (s, 2H), 8.05 (s , 8.17 (s, IH), 8.21 (s, IH), 8.89 (s, IH). 34

Pyrazolo [3,4-c] pyridine-3-amine < EMI ID = ^{_{1 H NMR (CD 3 OD,}} 300HMz): δ6.88 (d, J = 6.6 Hz, 1H), 7.28 (t, J = 8.2 Hz, 1H), 7.51 (m, 1H), 7.86 (m, 1H) , 8.19 (s, 2H), 8.48 (s, IH), 8.96 (s, IH)

The chemical structures of the compounds prepared in Examples 1-34 are summarized in Table 4 below.

Example constitutional formula Example constitutional formula One

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

< Experimental Example 1 > N-aryl-1H- Pyrazolopyridine -3- Amine Derivative MELK Evaluation of expression inhibition ability

In order to evaluate the ability of the N-aryl-1H-pyrazolopyridin-3-amine derivative according to the present invention to inhibit MELK expression, the following experiment was conducted.

Biotin labeled peptide substrate S1, HTRFRKinEASETM-TK kit (kit, cisbio) and Serine / Theronine (STK) monoclonal antibody were used to prepare the MELK of Example compound according to the present invention Expression-suppressing ability was measured.

To determine the degree of phosphorylation of biotin-labeled peptides, a homogeneous time-resolved fluorescence technology (HTRF) technique was used to measure TR-FRET (Time-Resolved Fluorescence Energy Transfer) signals. The test compound was dissolved in DMSO (dimethylsulfoxide) at 10 mM and then diluted to a DMSO concentration of 1%. The diluted compounds, MELK and Biotin labeled peptides (1 μM) were preincubated for 15 min at 25 ° C. in reaction buffer (HTRF kit 5 × buffer-cisbio, 1 M MgCl 2, 1 M Dithiothreitol DTT) adenosine triphosphate) was added thereto, followed by reaction at 25 ° C for 30 minutes. Streptavidin-XL665 (streptavidin) diluted in a detection buffer (HTRF kit, cisbio) containing ethylenediaminetetraacetic acid (EDTA), which serves to stop the kinase activity reaction, -XL665) and an ST3 anti-body labeled with Eu3 ⁺ -cryptate. After 1 hour incubation at 25 ° C, fluorescence polarization was measured with a multi-label reader (Wallac EnVision 2103, PerkinElmer). IC ₅₀ (half maximal inhibitory concentration) values were calculated using Prism (version 5.01, Graphpad Software, Inc.), and the results are shown in Table 5 below.

Example MELK IC ₅₀ (μM) One 1.864 2 5.122 3 6.873 4 11.223 5 12.542 6 25.893 7 26.542 8 22.342 9 21.872 10 8.921 11 87.54 12 > 100 13 57.32 14 > 100 15 98.547 16 > 100 17 0.356 18 1.529 19 19.723 20 21.342 21 19.712 22 12.730 23 9.125 24 8.743 25 9.718 26 11.324 27 5.724 28 19.873 29 15.324 30 12.343 31 0.331 32 1.200 33 14.532 34 94.56

As shown in Table 5,

The compounds of Examples according to the present invention inhibited MELK expression, and it was confirmed that the compounds of Examples 1, 17, 18, 31 and 32 inhibited MELK expression at a low IC ₅₀ value of 2 μM or less, It was found that the compounds of Examples 17 and 31 had an IC ₅₀ value of 0.4 μM or less, which suppresses the expression of MELK excellently even at a very low concentration.

Therefore, the compound represented by the formula (1) according to the present invention has an excellent effect of inhibiting the expression of MELK, so that it can prevent or treat MELK-related diseases such as breast cancer, lung cancer, bladder cancer, lymphoma, This can be useful.

&Lt; Formulation Example 1 > Preparation of powders

The compound represented by the formula (1) 2g

Lactose 1g

The above components were mixed and packed in airtight bags to prepare powders.

&Lt; Formulation Example 2 > Preparation of tablet

The compound represented by the formula (1) 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; Formulation Example 3 > Preparation of capsules

The compound represented by the formula (1) 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; Formulation Example 4 > Preparation of injection

The compound represented by the formula (1) 100 mg

Mannitol 180 mg

Na ₂ HPO ₄ .2H ₂ O 26 mg

Distilled water 2974 mg

According to the conventional method for preparing an injectable preparation, an injectable preparation was prepared by incorporating the aforementioned components in the amounts indicated.

&Lt; Formulation Example 5 > Preparation of health food

The compound represented by the formula (1) 500ng

Vitamin mixture Suitable amount

Vitamin A acetate 70 mg

Vitamin E 1.0 mg

vitamin 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 mg

Vitamin C 10 mg

Biotin 10 mg

Nicotinic acid amide 1.7 mg

Folic acid 50 mg

Calcium pantothenate 0.5 mg

Mineral mixture Suitable amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Claims

Claims 1. A compound represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
X, Y and Z are each independently carbon or nitrogen;
HetAr is 5- to 8-membered heteroarylene comprising at least one heteroatom selected from the group consisting of N, O and S;
R ¹ is hydrogen, straight or branched C _1- ₅ alkyl or unsubstituted N, O and S, and 5-8 membered heterocycloalkyl of atoms including a hetero atom at least one member selected from the group consisting of;
R ² is hydrogen or unsubstituted or substituted C _6- ₁₀ aryl, wherein the substituted C _6- ₁₀ aryl is optionally substituted with one or more substituents selected from the group consisting of C _1- ₅ alkoxy halogen and straight-chain or branched &Lt; / RTI >
R ³ is one or more substituents selected from hydrogen or straight chain or C _1- ₅ alkyl, C _1- ₅ alkoxy and the group consisting of halogen in the side chain;
R ⁴ is hydrogen or linear or branched C _1- ₅ alkyl; And
and n is an integer of 0-5).

The method according to claim 1,
X, Y and Z are each independently carbon or nitrogen;
HetAr is a 5-6 membered heteroarylene comprising at least one N atom;
R ¹ is a heterocycloalkyl of 5 to 6 atoms containing one or more C _1- ₃ alkyl or an unsubstituted N atom of hydrogen, linear or branched;
R ² is hydrogen or an unsubstituted or substituted _6- C ₈ aryl, wherein the substituted _6- C ₈ aryl is substituted with one or more substituents selected from the group consisting of C _1- ₃ alkoxy and halogen straight-chain or branched &Lt; / RTI >
R ³ is one or more substituents selected from hydrogen or straight chain or C _1- ₃ alkyl or C _1- ₃ alkoxy and the group consisting of halogen in the side chain;
R ⁴ is hydrogen or linear or branched C _1- ₃ alkyl; And
and n is an integer from 1 to 3; or a pharmaceutically acceptable salt thereof.

The method according to claim 1,
X, Y and Z are each independently carbon or nitrogen;
HetAr is pyrrolylene, pyrazolylene, pyridinylene or pyrazinylene;
R ¹ is hydrogen, methyl, ethyl, propyl, pyrrolidinyl or piperidinyl;
R ² is hydrogen or unsubstituted or substituted phenyl, wherein said substituted phenyl may be substituted with one or more substituents selected from the group consisting of fluoro, chloro, bromo, methoxy and ethoxy;
R ³ is hydrogen or one or more substituents selected from the group consisting of fluoro, chloro, bromo, methyl, ethyl, methoxy and ethoxy;
R < ⁴ > is hydrogen, methyl, ethyl or propyl; And
and n is an integer of 1 to 2, or a pharmaceutically acceptable salt thereof.

The method according to claim 1,
X, Y and Z are each independently carbon or nitrogen;
R ¹ -HetAr is

,

,

,

or

ego;
R < ¹ > is hydrogen, methyl or

ego;
R ² is hydrogen,

or

ego;
R ³ is hydrogen or one or more substituents selected from the group consisting of fluoro, chloro, methyl and methoxy;
R < ⁴ > is hydrogen or methyl; And
lt; / RTI > wherein n is 1, or a pharmaceutically acceptable salt thereof.

The method according to claim 1,
The compound represented by the formula (1) is any one selected from the group consisting of the following compounds, an optical isomer thereof or a pharmaceutically acceptable salt thereof:
(1) 1- (2-Chlorobenzyl) -N- (3-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(2) l- (2-Chlorobenzyl) -N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(3) l- (2-Chlorobenzyl) -N- (2-chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(4) Synthesis of 1- (2-chlorobenzyl) -N- (2,3-dichlorophenyl) -5- (pyridin- ;
(5) l- (2-Chlorobenzyl) -N- (3-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(6) l- (2-Chlorobenzyl) -N- (4-fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(7) 1- (2-chlorobenzyl) -N- (5-fluoro-2-methylphenyl) Amine;
(8) l- (2-Chlorobenzyl) -N- (2,5-dimethylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(9) 1- (2-Chlorobenzyl) -N- (3,5-dimethoxyphenyl) -5- (pyridin- 3- yl) -lH- pyrazolo [3,4- b] Amine;
(10) 1- (2-Chlorobenzyl) -N- (4-chlorophenyl) -N-methyl-5- (pyridin- Amine;
(11) l- (2-Chlorobenzyl) -N- (pyridin-2-yl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(12) 1- (3-Methoxybenzyl) -N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(13) N- (2-Chlorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(14) N- (3-Chlorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(15) N- (3-Fluorophenyl) -1- (3-methoxybenzyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(16) N- (5-fluoro-2-methylphenyl) -1- (3-methoxybenzyl) -5- - amine;
(17) To a solution of N- (3-chlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol- 3-amine;
(18) N- (3-Chlorophenyl) -5- (1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(19) N- (3-Chlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(20) N- (3,4-Dichlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(21) N- (3,5-Dichlorophenyl) -5- (1 -methyl-1H-pyrazol-4-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(22) Synthesis of N- (3,5-dichlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol- ] Pyridine-3-amine;
(23) N-phenyl-5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(24) N- (2-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(25) N- (3-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(26) N- (3-Fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(27) N- (4-Fluorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(28) N- (5-Fluoro-2-methylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(29) N- (2,5-Dimethylphenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-b] pyridin-3-amine;
(30) N- (3-Chlorophenyl) -5- (pyridin-3-yl) -1H-pyrazolo [3,4-c] pyridin-3-amine;
(3-chlorophenyl) -5- (1- (piperidin-4-yl) -lH-pyrazol-4-yl) -lH- pyrazolo [3,4- c] pyridine- 3-amine;
(32) To a solution of N- (3,4-dichlorophenyl) -5- (1- (piperidin-4-yl) ] Pyridine-3-amine;
(33) To a solution of N- (3,5-dichlorophenyl) -5- (1- (piperidin-4-yl) -1H-pyrazol- ] Pyridine-3-amine; And
(34) N- (3-Chlorophenyl) -5- (lH-pyrazol-4-yl) -lH-pyrazolo [3,4-c] pyridine-3-amine.

As shown in Scheme 1 below,
Reacting a compound represented by the formula (2) with a compound represented by the formula (3) to obtain a compound represented by the formula (4) (step 1);
A process for producing a compound represented by formula (1) as set forth in claim 1, comprising the step of reacting a compound represented by the formula (4) obtained in the above step 1 with a compound represented by the formula (5) to obtain a compound represented by the formula (1)
[Reaction Scheme 1]

(In the above Reaction Scheme 1,
A ¹ is fluoro, chloro or bromo, A ² is chloro, bromo or iodo, A ¹ is fluoro, A ² is chloro, bromo or iodo and A ¹ is chloro, A ² is bromo or iodo; when A ¹ is bromo, A ² is iodo;
D is

or

ego; And
Wherein X, Y, Z, R ¹ , R ² , R ³ , R ⁴ , HetAr and n are as defined in Chemical Formula 1 of claim 1.

The method according to claim 6,
The compound represented by the general formula (2)
Reacting a compound represented by the formula (6) with hydrazine hydrate to obtain a compound represented by the formula (7) (step 1);
Reacting a compound represented by the formula (7) obtained in the above step 1 to obtain a compound represented by the formula (8) (step 2);
Reacting the compound represented by the formula (8) with the compound represented by the formula (9) to obtain the compound represented by the formula (10) (step 3);
Reacting a compound represented by the formula (10) obtained in the above step 3 with a compound represented by the formula (11) to obtain a compound represented by the formula (12) (step 4); And
(Step 5) of reacting a compound represented by the formula (12) obtained in the step 4 with a compound represented by the formula (13) to obtain a compound represented by the formula (2).
[Reaction Scheme 2]

(In the above Reaction Scheme 2,
A ¹ is fluoro, chloro or bromo, A ² is chloro, bromo or iodo, A ¹ is fluoro, A ² is chloro, bromo or iodo and A ¹ is chloro, A ² is bromo or iodo; when A ¹ is bromo, A ² is iodo;
A < ³ > is halogen;
L ¹ is a C _1- ₃ alkoxy or halogen straight-chain or branched; And
R ² and n are the same as defined in the formula (1).

The method according to claim 6,
The compound represented by the general formula (2)
Reacting a compound represented by the formula (14) with sodium nitrite to obtain a compound represented by the formula (15) (step 1);
Reacting a compound represented by the formula (15) obtained in the above step 1 with a compound represented by the formula (9) to obtain a compound represented by the formula (12) (step 2); And
Reacting a compound represented by the formula (12) obtained in the step 2 with a compound represented by the formula (13) to obtain a compound represented by the formula (2)
[Reaction Scheme 3]

(In the above scheme 3,
A ¹ is fluoro, chloro or bromo, A ² is chloro, bromo or iodo, A ¹ is fluoro, A ² is chloro, bromo or iodo and A ¹ is chloro, A ² is bromo or iodo; when A ¹ is bromo, A ² is iodo;
A < ³ > is halogen; And
R ² and n are the same as defined in the formula (1).

A pharmaceutical composition for preventing or treating a MELK-related disease comprising the compound represented by the general formula (1) of claim 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

10. The method of claim 9,
The pharmaceutical composition for preventing or treating cancer, wherein the MELK-related disease is cancer.

11. The method of claim 10,
Wherein said cancer is any one selected from the group consisting of breast cancer, lung cancer, bladder cancer, lymphoma, polymorphic glial hyperplasia, and cervical cancer.

A health functional food composition for preventing or ameliorating a MELK-related disease, which comprises the compound represented by the general formula (1) of claim 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.