KR20170017607A - Novel 3-(isoxazol-3-yl)-pyrazolo[3,4-d]pyrimidin-4-amine compounds as RET kinase inhibitor - Google Patents

Abstract

The present invention relates to a novel 3-(isoxazol-3-yl)-pyrazolo[3,4-D]pyrimidine-4-amine compound, a manufacturing method, and medical uses for preventing and treated various cancer diseases caused by RET kinase activity of the compound.

Description

(Isoxazol-3-yl) -pyrazolo [3,4-d] pyrimidin-4-amine compound as a RET kinase inhibitor. d] pyrimidine-4-amine compounds as RET kinase inhibitor}

The present invention relates to a novel 3- (isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin-4-amine compound, a method for producing the compound, and various cancer diseases caused by RET kinase activity And to the use of the medicament for the prophylaxis and treatment of such diseases.

Rearranged during transfection Kinase is one of the receptor tyrosine kinases (RTKs) necessary for the development of sympathetic and parasympathetic nerves. It has been reported that the RET gene causes various types of thyroid cancer (MEN2A, MEN2B, FMTC, PTC, etc.) in the case of abnormal function such as point mutation, chromosome inversion, gene amplification. Among the thyroid cancer, MEN2A, MEN2B, and FMTC are caused by point mutation of RET gene, and PTC is reported to be caused by chromosomal translocation of RET gene. [Non-Patent Document 1. J. Med. Chem . 2012 , 55 , 4872-4876]

Recently, RET kinase has been attracting attention as a lung cancer inducer. [Non-patent document 2: Genome Res 2012 , 22 , 436-445] The expression of the fusion type tyrosine kinase KIF5B-RET or CCDC6-RET, in which the RET gene fuses with the KIF5B gene or the CCDC6 gene, . It has also been reported that NCOA4-RET and TRIM33-RET, a fusion type tyrosine kinase fused with the NCOA4 or TRIM33 (tripartite motif-containing 33) gene, are present in non-small cell lung cancer patients. [Non-Patent Document 3: Cancer Discov 2013 3 (6), 630-635]

Therefore, a compound having an inhibitory action of RET kinase is very useful for the prevention and treatment of various cancers.

Soretenib, Sunitinib, XL184, Vandetanib, and Ponatinib have been known as RET tyrosine kinase inhibitors, and they have been shown to inhibit cell proliferation of KIF5B-RET-expressing cell lines Inhibitory activity is reported. In addition, Cabozantinib, a RET tyrosine kinase inhibitor, has been reported to have a partial effect on non-small cell lung cancer caused by RET fusion gene expression.

On the other hand, it has been reported that compounds having a pyrazolo [3,4- d ] pyrimidin-4-amine parent nucleus exhibit an excellent inhibitory effect on various protein kinases. As an example thereof, International Patent Publication No. WO 2013/010136 (Patent Document 1) discloses a compound represented by the following formula (A).

(A)

Patent Document 1 Btk (Bruton's Tyrosine Kinase) having inhibitory effects with respect to protein-pyrazolo [3,4- d] the invention relates to a compound of pyrimidin-4-amine series, pyrazolo [3,4- d] pyrimidin Amin-4-amine is substituted with various aryl or heteroaryl groups at the C3 position, and as the heteroaryl group, isoxazol-5-yl group is substituted at the C3 position of the mother nucleus, The chemical structure of the substituted compound may also be described. However, Patent Document 1 confirms the synthesis and Btk inhibitory activity of a compound in which a phenoxyphenyl group is substituted at the C3 position of the pyrazolor [3,4- d ] pyrimidin-4-amine parent nucleus and is effective for the treatment of lymphoma tumor I can not tell you. That is, in Patent Document 1, only the chemical structure is disclosed for the compound in which the isoxazol-5-yl group is substituted at the C3 position of the pyrazolo [3,4- d ] pyrimidin- And no specific synthesis examples of the compound represented by the above formula (A) and the effect of inhibiting the Btk activity are disclosed at all.

On the other hand, the 3- (isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin-4-amine compound of the present invention is characterized in that the isoxazol- It may be optionally substituted, wherein the C5 position of the isoxazole-3-yl ring has - a (CH _{2) n} -A (wherein a is alkyl, aryl or heteroaryl group, n is an integer of 0 to 4) is substituted with , A novel compound whose chemical structure has not been known to date. In addition, the inventors of the present invention have found that the novel compounds of the present invention have an effect of inhibiting the activity of the RET gene among protein kinases, and have been used for the treatment of thyroid cancer, non-small cell lung cancer, It has excellent medicinal use.

International Patent Publication No. WO 2013/010136 "Inhibitor of Bruton Type Tyrosine Kinase (Btk)"

 Preparation of 3-substituted-1-isopropyl-1H-pyrazolo [3,4-d] pyrimidin-4-amines as RET Kinase Inhibitors ", J. Med. Chem. 2012, 55, 4872-4876  "A transformed KIF5B and RET gene fusion in lung adenocarcinoma revealed from whole-genome and transcriptome sequencing", Genome Res 2012, 22, 436-445  &Quot; Response to Cabozantinib in Patients with RET Fusion-Positive Lung Adenocarcinomas ", Cancer Discov 2013, 3 (6), 630-635

Accordingly, an object of the present invention is to provide a novel compound having excellent inhibitory activity and selectivity against RET kinase among protein kinases.

Another object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of cancers caused by abnormal expression of RET kinase containing the novel compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a process for preparing the novel compounds.

Another object of the present invention is to provide an intermediate compound useful in the synthesis of the novel compounds.

In order to solve the above problems, the present invention provides a pharmaceutical composition comprising 3- (isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin- A salt thereof, a hydrate thereof, a solvate thereof, and an isomer thereof.

[Chemical Formula 1]

In Formula 1,

는 단일결합 또는 이중결합을 나타내고;

Represents a single bond or a double bond;

A is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; A C ₆ -C ₁₅ aryl group; Or a five to six-membered heteroaryl group containing 1 to 2 hetero atoms selected from S and N;

R is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; C ₁ ~C ₁₀ hydroxyalkyl group; Benzyl group; Or a five to six-membered saturated or partially saturated heterocycloalkyl group containing from one to two heteroatoms selected from O and N;

n represents an integer of 0 to 4;

Wherein said aryl or heterocycloalkyl group is optionally substituted with at least one substituent selected from the group consisting of halo, C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, R ¹ -C (O) - wherein R ¹ is C ₁ -C ₁₀ alkyl or C ₂ -C ₁₀ alkenyl), or R ² -S (O) ₂ -, wherein R ² is C ₁ -C ₁₀ alkyl.

Since the 3- (isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin-4-amine compound according to the present invention exhibits an excellent inhibitory activity against RET protein kinase, For the purpose of preventing or treating a disease caused by a disease.

The diseases caused by abnormal cell growth in the present invention may include, for example, cancer diseases such as thyroid cancer, lung cancer and breast cancer.

The pharmaceutically acceptable salts of the compound represented by the formula (1) according to the present invention can be prepared by a conventional method in the art. Pharmacologically acceptable salts should be low in toxicity to humans and should not adversely affect the biological activity and physicochemical properties of the parent compound. Pharmaceutically acceptable salts include pharmaceutically acceptable free acids, acid addition salts of base compounds of formula (I), and alkali metal salts (such as sodium salts) and alkaline earth metal salts (such as calcium salts) Organic base addition salts of acids, and amino acid addition salts. The free acids that can be used in the preparation of pharmaceutically acceptable salts can be divided into inorganic and organic acids. As the inorganic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, bromic acid and the like can be used. The organic acid may be selected from the group consisting of acetic acid, methanesulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, fumaric acid, maleic acid, malonic acid, phthalic acid, succinic acid, lactic acid, citric acid, citric acid, gluconic acid, tartaric acid, Benzoic acid, embonic acid, aspartic acid, glutamic acid and the like can be used. Organic bases that can be used to prepare organic base addition salts include tris (hydroxymethyl) methylamine, dicyclohexylamine, and the like. Amino acids that can be used in the production of amino acid addition bases are natural amino acids such as alanine and glycine.

The compounds represented by Formula 1 according to the present invention include all hydrates and solvates as well as the pharmaceutically acceptable salts described above. The above-mentioned pharmaceutically acceptable salts can be prepared by a conventional method. For example, the base compound of Formula 1 may be dissolved in a solvent which can be mixed with water such as methanol, ethanol, acetone, 1,4-dioxane Followed by melting, followed by addition of a free acid or a free base, followed by crystallization or recrystallization.

In addition, the compound represented by Formula 1 according to the present invention may have one or more asymmetric centers, and in the case of such a compound, an enantiomer or diastereomer may exist. Accordingly, the present invention includes each isomer or a mixture of these isomers. The different isomers may be separated or cleaved by conventional methods, or any desired isomer may be obtained by conventional synthetic methods or by stereospecific or asymmetric synthesis.

The present invention also includes a radioactive derivative of the compound represented by Formula 1 according to the present invention, and these radioactive compounds are useful in the field of biomedical research.

The substituents used to define the compounds according to the invention are described in more detail below.

In the present invention, the term "halo" or "halogen atom" refers to chloro, fluoro, bromo, or iodo, which can be used interchangeably.

'Alkyl' in the present invention means a straight, branched or cyclic aliphatic saturated hydrocarbon group having 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms . Specific examples of these alkyl groups, methyl group, ethyl group, normal propyl group, an isopropyl group, a cyclopropyl group, a cyclopropyl group, a normal butyl group, isobutyl group, tert - butyl group, cyclobutyl group, n-pentyl group, iso Pentyl group, neopentyl group, tert -pentyl group, cyclopentyl group, normal hexyl group, isohexyl group, cyclohexyl group, normal heptyl group, n-octyl group and the like.

The term "hydroxyalkyl" in the present invention means an aliphatic saturated hydrocarbon group in which at least one hydroxyl group (-OH) is bonded to a straight chain or branched chain carbon chain as defined above. Specific examples of such a hydroxyalkyl group include a hydroxymethyl group, a 2-hydroxyethyl group, a 3-hydroxypropyl group, a 1-hydroxy-isopropyl group, a 2-hydroxybutyl group, 2-methyl-2-hydroxypropyl group and the like.

'Heterocycloalkyl' in the present invention means a saturated or partially saturated five to six-membered aliphatic ring group containing 1 to 2 hetero atoms selected from O and N. Specific examples of such a heterocycloalkyl group include a tetrahydrofuranyl group, a 2,3-dihydrofuranyl group, a 2,5-dihydrofuranyl group, a pyrrolidinyl group, a 2,3-dihydropyrrolidinyl group, Dihydropyrrolylinyl group, tetrahydro- 2H -pyranyl group, 3,4-dihydro- 2H -pyranyl group, 4H -pyranyl group, piperidinyl group, 1,2,3,4- Tetrahydropyridinyl group, 1,4-dihydropyridinyl group, piperazinyl, N -protected piperazinyl, morpholino group and the like. The N -protecting group of the piperazinyl may ordinarily include an alkyl group.

The term " aryl " in the present invention means a single ring, double ring, or three ring aromatic hydrocarbon group having 6 to 15 carbon atoms. Specific examples of such an aryl group include a phenyl group, a naphthyl group, an anthracenyl group, a phenanthrenyl group and the like.

'Heteroaryl' in the present invention means an aromatic ring group of 1 to 2 hetero atoms selected from the group consisting of S and N, and a single ring, a nugget or a tricyclic ring having 4 to 13 carbon atoms. These heteroaryls may be substituted with at least one substituent selected from the group consisting of a pyrrolyl group, a pyrazolyl group, an imidazolyl group, a thiazolyl group, an isothiazolyl group, a pyridinyl group, a pyrazinyl group, a pyridazinyl group, a pyrimidinyl group, An imidazolyl group, a benzimidazolyl group, a benzothiazolyl group, a benzisothiazolyl group, a quinolinyl group, an isoquinolinyl group, a phthalazinyl group, a quinazolinyl group and the like.

The compound according to the present invention can be represented by the following formula (1).

[Chemical Formula 1]

In Formula 1,

는 단일결합 또는 이중결합을 나타내고;

Represents a single bond or a double bond;

A is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; A C ₆ -C ₁₅ aryl group; Or a five to six-membered heteroaryl group containing 1 to 2 hetero atoms selected from S and N;

R is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; C ₁ ~C ₁₀ hydroxyalkyl group; Benzyl group; Or a five to six-membered saturated or partially saturated heterocycloalkyl group containing from one to two heteroatoms selected from O and N;

n represents an integer of 0 to 4;

Wherein said aryl or heterocycloalkyl group is optionally substituted with at least one substituent selected from the group consisting of halo, C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, R ¹ -C (O) - wherein R ¹ is C ₁ -C ₁₀ alkyl or C ₂ -C ₁₀ alkenyl), or R ² -S (O) ₂ -, wherein R ² is C ₁ -C ₁₀ alkyl.

In the compound represented by the above formula (1), it is preferable that A is a hydrogen atom; A tert -butyl group, a cyclobutyl group, a n-pentyl group, an isopentyl group, a neopentyl group, a tert -butyl group, a tert -butyl group, a tert- An alkyl group selected from a pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A phenyl group substituted or unsubstituted with substituents selected from halo, methyl, ethyl, normal propyl, isopropyl, cyclopropyl, methoxy and ethoxy; A thiophene group; Or a pyridinyl group.

In the compound represented by the general formula (1), R is preferably a hydrogen atom; A tert -butyl group, a cyclobutyl group, a n-pentyl group, an isopentyl group, a neopentyl group, a tert -butyl group, a tert -butyl group, a tert- An alkyl group selected from a pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A hydroxyalkyl group selected from a 2-methyl-2-hydroxypropyl group, a 2-hydroxybutyl group and a 4-hydroxybutyl group; Benzyl group; Tetrahydro-2H - pyranyl group; Or a piperidinyl group substituted or unsubstituted with R ¹ -C (O) - or R ² -S (O) ₂ -, wherein R ¹ is a C ₁ -C ₆ alkyl group or a C ₂ -C ₆ alkenyl group , R ² is a compound representing a C ₁ ~C ₆ alkyl group.

는 이중결합을 나타내고; 상기 A는 메틸기, 에틸기, 노말프로필기, 이소프로필기, 사이클로프로필기, 치오펜-2-일기 또는 치오펜-2-일기를 나타내고; 상기 R은 메틸기, 에틸기, 노말프로필기, 이소프로필기, 사이클로프로필기, 사이클로프로필메틸기, 노말부틸기, 이소부틸기, tert-부틸기, 사이클로부틸기, 노말펜틸기, 이소펜틸기, 네오펜틸기, tert-펜틸기, 사이클로펜틸기, 노말헥실기, 이소헥실기, 사이클로헥실기 및 노말헵틸기로부터 선택된 알킬기; 2-메틸-2-하이드록시프로필기, 2-하이드록시부틸기 및 4-하이드록시부틸기로부터 선택된 하이드록시알킬기; 벤질기; 또는 테트라하이드로-2H-파이란일기를 나타내고; 상기 n은 0을 나타내는 나타내는 화합물이다.In the compound represented by Formula 1, particularly preferably,

Represents a double bond; A represents a methyl group, an ethyl group, a normal propyl group, an isopropyl group, a cyclopropyl group, a thiophen-2-yl group or a thiophen-2-yl group; Wherein R is a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a cyclopropyl group, a cyclopropyl group, a normal butyl group, isobutyl group, tert - butyl group, cyclobutyl group, n-pentyl group, isopentyl group, neopentyl A tert -pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A hydroxyalkyl group selected from a 2-methyl-2-hydroxypropyl group, a 2-hydroxybutyl group and a 4-hydroxybutyl group; Benzyl group; Or a tetrahydro-2H - pyranyl group; And n represents 0.

The compound represented by the formula (1) is more specifically exemplified as follows:

1-Isopropyl-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 1);

1-Isopropyl-3- (5-cyclopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 2);

1-Isopropyl-3- (5-isopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 3);

3- (5- ( tert -Butyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 4);

3- (5-Cyclopentylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 5);

3- (5-Cyclohexylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 6);

3-yl) -1 H- pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 7);

L-isopropyl-3- (5- (thiophen-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 8);

3- (5- (4-Fluorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 9);

L-isopropyl-3- (5- (4-methoxyphenyl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 10);

3- (5- (3-Chlorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 11);

3- (5- (2-Chlorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 12);

L-isopropyl-3- (5- (pyridin-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 13);

L-isopropyl-3- (5- (pyridin-3-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 14);

3- (5-benzylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 15);

1-isopropyl-3- (5- (phenyl-4,5-hydroxyimino-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine (Compound No. 16 );

3- (5-phenyl-3-yl) -1- (tetrahydro -2 H- Failan-4-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine ( Compound No. 17);

1- (1- (methylsulfonyl) piperidin-4-yl) -3- (5- (phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin- 4-amine (Compound No. 18);

1- (4- (4-amino-3- (5- (phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1-yl) piperidin-yl ) Ethanone (Compound No. 19);

(±) -1- (3- (4- amino-3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1-yl) piperidine 1-yl) prop-2-en-1-one (Compound No. 20);

(4-amino-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- 1-yl) prop-2-en-1-one (Compound No. 21);

3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 22);

L -Cyclohexyl-3- (5-cyclopropylisoxazol-3-yl) -1H-pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 23);

3- (5-cyclopropyl-a-3-yl) -1- (tetrahydro -2 H - Failan-4-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 24);

3- (5-Cyclopropylisoxazol-3-yl) -1-heptyl-1 H -pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 25);

3- (5-Cyclopropylisoxazol-3-yl) -1-methyl- 1H -pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 26);

L -Benzyl-3- (5-cyclopropylisoxazol-3-yl) -1H-pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 27);

1- (4-Amino-3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-1-yl) -2-methyl-propan-2 Ol (Compound No. 28);

3- (5-Ethylisoxazol-3-yl) -1-isopropyl- 1H -pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 29);

1-Isopropyl-3- (isoxazol-3-yl) -1 H -pyrazolo [3,4- d ] pyrimidin-4-amine (Compound No. 30).

The present invention also provides a process for producing the compound represented by the above formula (1). The production method according to the present invention will be described in detail as follows.

Production Method 1

According to the preparation method according to the following Reaction Scheme 1, the compound represented by Formula 1 can be prepared by performing a four step process using a compound represented by Formula 2 below as a starting material.

 [Reaction Scheme 1]

, A, R 및 n은 각각 상기 화학식 1에서 정의한 바와 같다)(In the above Reaction Scheme 1,

, A, R and n are each as defined in the above formula (1)

The production method according to Reaction Scheme 1 will be described in detail below for each reaction.

I) The reaction is carried out by reacting 3-bromo-1 H- pyrazolo [3,4- d ] pyrimidin-4-amine and RX (wherein X represents a halogen atom) To thereby prepare a compound represented by the above formula (3) in which various R's are introduced. The reaction can be carried out under the condition that i) the reaction is heated to 70 to 80 캜 in the presence of an inorganic base. In this case, the inorganic base may be selected from hydroxides, oxides, carbonates, and sulfates of alkali metals or earth metals, and preferably an alkali metal carbonate such as potassium carbonate may be used. As the reaction solvent, a conventional organic solvent can be used. In the examples of the present invention, dimethylformamide (DMF) is mainly used. However, the solvent of the present invention is not limited thereto.

Ii) The reaction is a process for preparing the compound represented by Chemical Formula 4 in which the vinyl group is introduced into the compound represented by Chemical Formula 3 by Stille reaction. Stille reaction is carried out under reflux conditions using a metal catalyst such as palladium (Pd) or nickel (Ni) and an organotin compound as a coupling reagent. As the metal catalyst, tetrakis (triphenylphosphine) palladium [Pd (PPh ₃ ) ₄ ] can be typically used, and tributylvinyltin can be used as the organic tin compound. As the reaction solvent, aromatic hydrocarbons such as toluene may be used, and the reaction solvent may be in the range of about 120 ° C to 150 ° C as the reflux temperature of the used solvent.

Iii) The reaction is a process of converting the vinyl group of the compound of Formula 4 into an aldehyde group by oxdative cleavage and then converting it into an oxime group to produce the compound of Formula 5. [ The oxidative cleavage reaction to convert the vinyl group to an aldehyde group can proceed using ozone (O ₃ ). The oxidative cleavage reaction is carried out by carrying out a hydroxylation reaction using osmium tetroxide (OsO ₄ ) as a catalyst and N -methylmorpholine- N -oxide (NMO) as an oxidizing agent, and then sodium periodate ₄ ). ≪ / RTI > Then, the aldehyde compound is reacted with hydroxylamine (NH ₂ OH) in an ethanol solvent to introduce an oxime group.

(Iv) The reaction is a process for preparing the compound represented by the above formula (1) by a nitrile oxide cycloaddition reaction of the compound represented by the formula (5). The cyclization addition reaction may be carried out by using a hypervalent iodine compound such as (diacetoxy iodo) benzene or phenyl iodine bis (trifluoroacetate) as a reaction reagent and in a mixed solvent of alcohol and water At room temperature (20 캜 to 30 캜).

As a core intermediate compound synthesized during the production process according to Reaction Scheme 1, the compound having the vinyl functional group represented by Formula 4 and the compound having the oxime functional group represented by Formula 5 are each a novel compound. Therefore, the present invention is also characterized by an intermediate compound represented by the following general formula (4) or (5).

[Chemical Formula 4]

(Wherein R is a hydrogen atom, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ hydroxyalkyl group, a benzyl group, or a five to six-membered heterocyclic group containing 1 to 2 hetero atoms selected from O and N Saturated or partially saturated heterocycloalkyl group.

[Chemical Formula 5]

Wherein R is a hydrogen atom, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ hydroxyalkyl group, a benzyl group, or a five to six-membered heterocyclic group containing 1 to 2 hetero atoms selected from O and N Saturated or partially saturated heterocycloalkyl group.

The intermediate compound represented by Formula 4 may typically include 1-isopropyl-3-vinyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine.

The intermediate compound represented by Formula 5 may typically include ( E ) -4-amino-1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidine-3-carbaldehyde oxime.

Production Method 2

According to the production method according to the following Reaction Scheme 2, a compound represented by the following Formula 1 can be prepared by performing the three-step preparation process using the malononitrile compound represented by the following Formula 6 as a starting material.

[Reaction Scheme 2]

, A, R 및 n은 각각 상기 화학식 1에서 정의한 바와 같다)(In the above Reaction Scheme 2,

, A, R and n are each as defined in the above formula (1)

The production method according to Reaction Scheme 2 will be described in detail below for each reaction.

a) The reaction is carried out by cyclizing a malononitrile compound represented by the formula (6) and hydrazine (NH ₂ NH ₂ ) hydrate to prepare a 1 H -pyrazole-4-carbonitrile compound represented by the formula to be. The reaction a) may be carried out in an alcoholic solvent such as methanol, ethanol or the like at a room temperature (20 ° C to 30 ° C).

b) reacting the 1 H represented by the formula 7 by cyclization of the pyrazole-4-carbonitrile compound to form an amide compound _{(HC (O) NH 2)} , 1 H represented by the formula (8) -pyrazolo [3,4-d] pyrimidin-4-amine compound. The reaction b) may be carried out under a condition of heating from 130 ° C to 180 ° C.

c) The reaction is carried out by reacting the 1 H -pyrazolo [3,4-d] pyrimidin-4-amine compound represented by the above formula (8) with an alcohol compound represented by ROH, ≪ / RTI > The c) reaction proceeds through a Mitsunobu reaction using triphenylphosphine (PPh ₃ ) and an azodicarboxylate compound. The azodicarboxylate compound may include, for example, diethyl azodicarboxylate (DEAD) or diisopropyl azodicarboxylate (DIAD). The Mitsunobu reaction can be carried out at room temperature (20 ° C to 30 ° C) using a polar solvent such as tetrahydrofuran.

Meanwhile, the present invention includes a pharmaceutical composition comprising the compound represented by Formula 1, a pharmaceutically acceptable salt thereof, a solvate thereof, and a hydrate thereof as an active ingredient.

Since the compound represented by Formula 1 exhibits an excellent inhibitory activity against RET kinase, it can be used as a preventive or therapeutic agent for diseases caused by abnormal cell growth.

Cancer diseases caused by abnormal cell growth in the present invention may specifically include thyroid cancer, lung cancer, and breast cancer. In particular, it has an excellent effect of inhibiting the activity of RET kinase, and thus is useful as a prophylactic and therapeutic agent for thyroid carcinoma, non-small cell lung cancer and breast cancer.

The pharmaceutical composition of the present invention comprises a compound represented by the general formula (1), a pharmaceutically acceptable salt thereof, a solvate thereof, and a hydrate thereof as an active ingredient, wherein a usual non-toxic pharmaceutically acceptable carrier, adjuvant and excipient And the like can be formulated into preparations for oral administration such as tablets, capsules, troches, liquids and suspensions, or parenteral administration preparations which are customary in the pharmaceutical field.

Examples of excipients which can be used in the pharmaceutical composition of the present invention include sweeteners, binders, solubilizers, solubilizers, wetting agents, emulsifiers, isotonizing agents, adsorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, perfumes and the like. For example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, stearin, magnesium stearate, magnesium aluminum silicate, starch, gelatin, tragacanth gum, Water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavor, and the like.

The dose of the compound according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and disease severity of the patient, and is generally 0.01 To 1,000 mg / day. Depending on the judgment of a doctor or a pharmacist, it may be administered once to several times a day at a predetermined interval.

The present invention will now be described in more detail with reference to the following examples, preparation examples and experimental examples. However, the following examples, preparation examples and experimental examples are merely illustrative of the present invention, The present invention is not limited thereto.

EXAMPLES Synthesis of Compound (1)

Example 1. Synthesis of 1-isopropyl-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-

Step 1: 3-Bromo-l-isopropyl - lH - pyrazolo [3,4- d ] pyrimidin-

To a round bottom flask inside was added to 3-bromo -1- H- pyrazolo [3,4 -d] pyrimidin-4-amine (3 g, 14.02 mmol) and potassium carbonate (7.75 g, 56.1 mmol) And filled with nitrogen. To the filled flask was added dimethylformamide (93 mL), followed by 2-bromopropane (1.45 mL, 15.4 mmol). After stirring at 80 ° C for 12 hours, the reaction mixture was diluted with ethyl acetate and the reaction was terminated by using water. The reaction mixture was extracted with ethyl acetate and water. The obtained organic layer was washed with water several times, dried over anhydrous magnesium sulfate, and filtered. The resulting residue was purified by column chromatography (EtOAc: n- Hexane = 1: 2) to give the title compound (2.4 g, 67% .

^{1 H NMR (400 MHz, CDCl} 3) δ 8.32 (s, 1H), 5.84 (bs, 2H), 5.09 (septet, J = 6.8 Hz, 1H), 1.54 (d, J = 6.8 Hz, 6H)

Step 2: l-Isopropyl-3-vinyl - lH - pyrazolo [3,4- d ] pyrimidin-

To a round bottom flask was added 3-bromo-1-isopropyl -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine (3.99 g, 15.6 mmol) and _{Pd (PPh 3) 4 (3.6} g, 3.12 mmol), and the inside was filled with nitrogen. Toluene (156 mL) and tributylvinyltin (5.57 mL, 18.7 mmol) were added thereto, followed by reflux at 150 ° C for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and the solvent was distilled off under reduced pressure. To the residue was added tetrahydrofuran / 1 N hydrochloric acid (3/1, 160 mL) and the mixture was stirred at room temperature for 12 hours. After diluting with ethyl acetate, the organic layer was extracted several times with 1 N HCl, and the combined aqueous layers were neutralized with a sodium bicarbonate solution in a round bottom flask. The resulting residue was purified by column chromatography (EtOAc: n- Hexane = 1: 1) to give the title compound (2.59 g, 82% yield). MS: m / ≪ / RTI >

^{1 H NMR (400 MHz, CDCl} 3) δ 8.33 (s, 1H), 7.25 (s, 1H), 6.97 (dd, J = 11.2 Hz, 18.0 Hz, 1H), 5.93 (d, J = 18.0 Hz, 1H ), 5.65-5.62 (m, 3H), 5.10 (septet, J = 6.8 Hz, 1H), 1.54 (d, J = 6.8 Hz,

Step 3: ( E ) -4-Amino-l-isopropyl - lH - pyrazolo [3,4- d ] pyrimidine- 3-carbaldehyde oxime

3-vinyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine (2.07 g, 10.19 mmol) was added to a round bottom flask, and tetrahydrofuran / water , 140 mL). In subsequent 0 ℃ N - methyl morpholine - N - oxide (4.77 mL, 20.38 mmol, 50 w / w% in water) and osmium tetroxide _{(OsO 4; 6.5 mL, 1.02} mmol, 4 w / w% in H ₂ O). After stirring for 3 hours at room temperature, the reaction was terminated using sodium sulfite solution, and the aqueous layer was extracted several times with chloroform / isopropyl alcohol (4/1) solvent. The collected organic solvent was distilled under reduced pressure, and the residue was dissolved in tetrahydrofuran / water (3/1, 140 mL). Then, sodium periodate (NaIO ₄ ; 4.36 g, 20.38 mmol) was added thereto, followed by stirring at room temperature for 2 hours. After the reaction was completed, the suspension was filtered using celite, and ethanol (16 mL) was added to the obtained residue. Subsequently, sodium acetate (2 g, 24.46 mmol) and hydroxylamine hydrochloride (841 mg, 25.48 mmol) were added thereto, followed by stirring at room temperature for 12 hours. The solvent was distilled off under reduced pressure, and the residue was extracted with ethyl acetate and water. The obtained organic layer was washed with water several times, dried over anhydrous magnesium sulfate and filtered. The resulting residue was subjected to column chromatography (EtOAc: n- Hexane = 2: 1) to give the title compound (2 g, 89% .

^{1 H NMR (400 MHz, CDCl} 3) δ 8.39 (s, 1H), 8.31 (s, 1H), 8.17 (bs, 1H), 5.81 (bs, 1H), 5.12 (septet, J = 6.8 Hz, 1H) , 1.54 (d, J = 6.8 Hz, 6H)

Step 4: 1-Isopropyl-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-

Was added to a round bottom flask was added (E) -4- amino-1-isopropyl -1 H- pyrazolo [3,4 -d] pyrimidin-3-carbazole aldehyde oxime (220 mg, 1 mmol), methanol / And dissolved in a mixed solvent of water (8.0 mL / 1.6 mL). After the addition of thionylbenzene (0.15 mL, 1.1 mmol), phenyl iodine bis (trifluoroacetate) (PIFA) was added over 3 h (215 mg, 0.5 mmol each) over 2 h. Then, the mixture was stirred for 4 hours and extracted with ethyl acetate. The obtained organic layer was washed with water several times, dried over anhydrous magnesium sulfate, and filtered. The resulting residue was subjected to column chromatography (EtOAc: n- Hexane = 1: 2) to give the title compound (134 mg, 42% .

^{1 H NMR (400 MHz, CDCl} 3) δ 8.91 (bs, 1H), 8.32 (s, 1H), 7.88-7.86 (m, 2H), 7.54-7.26 (m, 3H), 7.19 (s, 1H), 6.91 (bs, 1H), 5.19 (septet, J = 6.8 Hz, 1H), 1.61 (d, J = 6.8 Hz, 6H)

Examples 2-16.

( E ) -4-Amino-1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidine-3-carbaldehyde oxime with a variety of diastereoisomers according to the method illustrated in step 4 of Example 1, Nitrile oxide cycloaddition was carried out using an acetylene compound to synthesize the respective title compounds.

Example 2. Synthesis of 1-isopropyl-3- (5-cyclopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.72 (bs, 1H), 8.27 (s, 1H), 6.86 (bs, 1H), 6.52 (s, 1H), 5.11 (sep, J = 6.8 Hz, 1H) , 2.11-2.04 (m, 1H), 1.53 (d, J = 6.8 Hz, 6H), 1.13-0.99 (m, 4H)

Example 3. Synthesis of 1-isopropyl-3- (5-isopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.66 (bs, 1H), 8.33 (s, 1H), 6.64 (d, J = 0.8 Hz, 1H), 5.88 (bs, 1H), 5.15 (sep, J = 6.8 Hz, 1H), 3.16 ( sep, J = 7.2 Hz, 1H), 1.58 (d, J = 6.8 Hz, 6H), 1.39 (d, J = 6.8 Hz, 6H)

Example 4. 3- (5- (tert - butyl) isoxazol-3-yl) -1-isopropyl -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine (Compound No. 4) Synthesis of

^{1 H NMR (400 MHz, CDCl} 3) δ 8.67 (bs, 1H), 8.33 (s, 1H), 6.63 (s, 1H), 5.85 (bs, 1H), 5.17 (sep, J = 6.8 Hz, 1H) , 3.16 (sep, J = 7.2 Hz, 1H), 1.58 (d, J = 6.8 Hz, 6H)

Example 5. Synthesis of 3- (5-cyclopentylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.67 (bs, 1H), 8.33 (s, 1H), 6.64 (s, 1H), 5.93 (bs, 1H), 5.15 (sep, J = 6.4 Hz, 1H) , 3.27 (quin, J = 7.2 Hz, 1H), 2.18-2.12 (m, 2H), 1.82-1.72 (m, 6H), 1.57 (d, J = 6.4 Hz,

Example 6. Synthesis of 3- (5-cyclohexylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.67 (bs, 1H), 8.33 (s, 1H), 6.62 (s, 1H), 5.94 (bs, 1H), 5.15 (sep, J = 6.8 Hz, 1H) , 2.85 (tt, J = 3.6 Hz, 10.8 Hz, 1H), 2.14-2.11 (m, 2H), 1.85 (dt, J = 3.6 Hz, 9.6 Hz, 2H), 1.77-1.73 (m, 1H), 1.58 (d, J = 6.8 Hz, 6H), 1.56-1.25 (m, 5H)

Example 7. Synthesis of 1-isopropyl-3- (5- (thiophen-3-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- No. 7)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.61 (bs, 1H), 8.35 (s, 1H), 7.87 (dd, J = 1.2 Hz, 2.8 Hz, 1H), 7.49 (dd, J = 1.2 Hz, 4.8 Hz, 1H), 7.46 (dd , J = 2.8 Hz, 4.8 Hz, 1H), 7.04 (s, 1H), 5.18 (sep, J = 6.4 Hz, 1H), 1.59 (d, J = 6.4 Hz, 6H)

Example 8. Synthesis of 1-isopropyl-3- (5- (thiophen-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- No. 8)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.57 (bs, 1H), 8.35 (s, 1H), 7.61 (dd, J = 0.8 Hz, 4.0 Hz, 1H), 7.52 (dd, J = 1.2 Hz, 5.2 Hz, 1H), 7.18 (dd , J = 4.0 Hz, 5.2 Hz, 1H), 5.87 (bs, 1H), 5.18 (sep, J = 6.8 Hz, 1H), 1.60 (d, J = 6.8 Hz, 6H)

Example 9 Synthesis of 3- (5- (4-fluorophenyl) isoxazol-3-yl) -l-isopropyl-lH - pyrazolo [3,4- d ] pyrimidin- 9) Synthesis of

^{1 H NMR (400 MHz, CDCl} 3) δ 8.59 (bs, 1H), 8.35 (s, 1H), 7.88-7.84 (m. 2H), 7.21 (t, J = 8.8 Hz, 2H), 7.14 (s, 1H), 7.02 (d, J = 8.8 Hz, 2H), 5.84 (bs, 1H), 5.18 (sep, J = 6.8 Hz, 1H), 1.59 (d, J = 6.8 Hz, 6H)

Example 10 1-Isopropyl-3- (5- (4-methoxyphenyl) isoxazol-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine (Compound No. 10)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.68 (bs, 1H), 8.35 (s, 1H), 7.81 (d, J = 8.4 Hz, 2H), 7.08 (s, 1H), 7.02 (d, J = J = 6.8 Hz, 1H), 3.89 (s, 3H), 1.60 (d, J = 6.4 Hz, 6H)

Example 11 3- (5- (3-Chlorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin- ) Synthesis of

^{1 H NMR (400 MHz, CDCl} 3) δ 8.55 (bs, 1H), 8.35 (s, 1H), 7.86 (s, 1H), 7.77-7.74 (m, 1H), 7.48-7.45 (m, 2H), (S, 1H), 5.90 (bs, 1H), 5.18 (sep, J = 6.8 Hz, 1H), 1.59 (d, J = 6.8 Hz,

Example 12. Synthesis of 3- (5- (2-chlorophenyl) isoxazol-3-yl) -l-isopropyl-lH - pyrazolo [3,4- d ] pyrimidin- ) Synthesis of

^{1 H NMR (400 MHz, CDCl} 3) δ 8.59 (bs, 1H), 8.36 (s, 1H), 7.99-7.98 (m, 1H), 7.58-7.56 (m, 2H) 7.48-7.42 (m, 2H) , 5.82 (bs, 1H), 5.18 (sep, J = 6.8 Hz, 1H), 1.61 (d, J = 6.8 Hz,

Example 13. Synthesis of l-isopropyl-3- (5- (pyridin-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- Synthesis of acetic acid salt (Compound No. 13)

^{1 H NMR (400 MHz, CDCl} 3) δ 9.67 (bs, 1H), 8.75-8.74 (m, 1H), 8.30 (s, 1H), 7.99-7.97 (m, 1H), 7.89 (td, J = 1.6 J = 6.8 Hz, 1H), 1.61 (d, J = 6.8 Hz, 6H), 7.57 (s, 1H), 7.43-7.39

Example 14 Synthesis of 1-isopropyl-3- (5- (pyridin-3-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- 14)

^{1 H NMR (400 MHz, CDCl} 3) δ 9.13 (dd, J = 0.8 Hz, 2.0 Hz, 1H), 8.73 (dd, J = 1.6 Hz, 4.8 Hz, 1H), 8.52 (bs, 1H), 8.36 ( (s, 1H), 8.16 (dt, J = 2.0 Hz, 8.0 Hz, 1H), 7.47 (ddd, J = 0.4 Hz, 4.8 Hz, 7.6 Hz, 1H) ), 5.19 (sep, J = 6.8 Hz, 1H), 1.60 (d, J = 6.8 Hz,

Example 15. Synthesis of 3- (5-benzylisoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.60 (bs, 1H), 8.31 (s, 1H), 7.39-7.26 (m, 5H), 6.61 (s, 1H), 5.81 (bs, 1H), 5.14 ( sep, J = 6.8 Hz, 1H ), 4.15 (s, 2H), 1.54 (d, J = 6.4 Hz, 6H)

Example 16. Synthesis of l-isopropyl-3- (5- (phenyl-4,5-dihydroxyisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- (Compound No. 16) Synthesis of

^{1 H NMR (400 MHz, CDCl} 3) δ 8.42 (bs, 1H), 8.32 (s, 1H), 7.41-7.34 (m, 5H), 5.94 (bs, 1H), 5.78 (dd, J = 8.8 Hz, J = 6.8 Hz, 1H), 3.98 (dd, J = 11.2 Hz, 17.6 Hz, 1H), 3.57 (dd, J = 8.4 Hz, 17.6 Hz, 1H), 1.52 t, J = 7.2 Hz, 6H)

Example 17. 3- (5-phenyl-3-yl) -1- (tetrahydro -2 H- Failan-4-yl) -1 H- pyrazolo [3,4 -d] pyrimidin- Synthesis of 4-amine (Compound No. 17)

Step 1: 2- (Methoxy (5-phenylisoxazol-3-yl) methylene) malononitrile

5-Phenylisoxazole-3-carboxylic acid (2 g, 10.57 mmol) was added to a round bottom flask and the inside was filled with nitrogen. Methylene chloride (35 mL) and a catalytic amount of dimethylformamide (5 drops) were added to the flask, and oxalyl chloride (4.53 mL, 52.85 mmol) was added slowly. After stirring at room temperature for 2 hours, the residue was dried under reduced pressure and used directly for the next reaction without purification.

Sodium hydride (845 mg, 21.14 mmol) and malononitrile (768 mg, 11.63 mmol) were added to a round-bottomed flask, which was filled with nitrogen and then tetrahydrofuran (35 mL) was added. After cooling to 0 ° C, the previously obtained residue was dissolved in a tetrahydrofuran solution (20 mL) and slowly added dropwise to the reaction solution. After stirring for 22 hours, diisopropylethylamine (7.3 mL, 42.28 mmol) and dimethyl sulfate (8 mL, 84.56 mmol) were added and the mixture was warmed to 90 ° C and stirred. After stirring for 12 hours, the reaction was terminated with water and extracted with ethyl acetate and water. The combined organic layers were washed several times with water, dried over anhydrous magnesium sulfate and filtered to give the title compound.

^{1 H NMR (400 MHz, CDCl} 3) δ 7.81-7.79 (m, 2H), 7.51-7.26 (m, 3H), 6.88 (s, 1H), 4.20 (s, 3H)

Step 2: 5-Amino-3- (5-phenylisoxazol-3-yl) -lH - pyrazole-4-carbonitrile

To the round bottom flask was added 2- (methoxy (5-phenylisoxazol-3-yl) methylene) malononitrile (2.66 g, 10.57 mmol), followed by ethanol (30 mL) at room temperature. Hydrazine hydrate (0.4 mL, 11.1 mmol) was added to the reaction solution, followed by stirring for 12 hours. Thereafter, water was added to terminate the reaction, followed by extraction with isopropyl alcohol / chloroform solution and drying with anhydrous magnesium sulfate. The resulting residue was washed several times with ethyl acetate and methylene chloride to give the title compound.

^{1 H NMR (400 MHz, DMSO} ) δ 12.61 (bs, 2H), 7.94 (dd, J = 2.0 Hz, 8.0 Hz, 2H), 7.57-7.52 (m, 3H), 7.29 (s, 1H), 6.60 ( bs, 1H)

Step 3: 3- (5-Phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-

(5-phenylisoxazol-3-yl) -1H - pyrazole-4-carbonitrile (220 mg, 0.88 mmol) and formamide (4 mL) were added to a round bottom flask , And the mixture was stirred at 160 DEG C for 6 hours. When a solid was gradually formed, the temperature was lowered to room temperature and water was added to terminate the reaction. The obtained solid was washed several times with water, chloroform and ethanol to obtain the title compound.

^{1 H NMR (400 MHz, DMSO} ) δ 14.04 (bs, 1H), 8.22 (s, 1H), 8.05 (bs, 2H), 8.00 (dd, J = 2.0 Hz, 8.0 Hz, 2H), 7.63 (s, 1H), < / RTI > 7.60-7.53 (m, 3H)

Step 4: 3- (5-phenyl-3-yl) -1- (tetrahydro -2 H- Failan-4-yl) -1 H- pyrazolo [3,4 -d] pyrimidin -4 - amine

(17 mg, 0.108 mmol) and 3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine (15 mg, 0.054 mmol) and triphenylphosphine (21 mg, 0.081 mmol) were added thereto, followed by dissolution in tetrahydrofuran (2 mL). Diisopropyl azodicarboxylate (DIAD; 0.002 mL, 0.081 mmol) was then slowly added. After stirring for 2 hours, water was added to terminate the reaction, followed by extraction with ethyl acetate. The collected organic layer was washed with water several times, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was subjected to column chromatography (EtOAc: n- Hexane = 2: 1) to obtain the title compound.

^{1 H NMR (400 MHz, CDCl} 3) δ 8.46 (bs, 1H), 8.34 (s, 1H), 7.88-7.84 (m, 2H), 7.53-7.47 (m, 3H), 7.16 (s, 1H), 5.81 (bs, 1H), 5.01 (tt, J = 4.0 Hz, 12.0 Hz, 1H), 4.15 (dd, J = 4.0 Hz, 11.2 Hz, 2H), 3.63 (td, J = 2.0 Hz, 12.0 Hz, 2H ), 2.42 (qd, J = 4.4 Hz, 12.4 Hz, 2H), 2.03-1.95 (m, 2H)

Examples 18-20.

On the basis of the method illustrated in Step 4 of Example 17, 3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-4-amine with various Mitsunobu reaction was carried out using an alcohol compound to synthesize the respective title compounds.

Example 18. 1- (1- (methylsulfonyl) piperidin-4-yl) -3- (5- (phenyl-3-yl) -1 H- pyrazolo [3,4 -d ] Pyrimidin-4-amine (Compound No. 18)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.68 (bs, 1H), 8.34 (s, 1H), 7.87-7.85 (m, 2H), 7.54-7.49 (m, 3H), 7.16 (s, 1H), 5.88 (bs, 1H), 4.92 (tt, J = 4.4 Hz, 11.6 Hz, 1H), 4.01 (d, J = 12.0 Hz, 2H), 2.99 (td, J = 2.4 Hz, 12.4 Hz, 2H), 2.87 (s, 3H), 2.47 (qd, J = 4.4 Hz, 12.0 Hz, 2H), 2.16-2.14

Example 19. l- (4- (4-Amino-3- (5- (phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- Ylidene) ethanone (Compound No. 19) Synthesis of

¹ H NMR (400 MHz, CDCl ₃ )? 8.66 (bs, IH), 8.35 (s, IH), 7.87-7.84 (m, 2H), 7.53-7.48 2H), 3.37-3.29 (m, 2H), 2.85 (m, 2H), 5.87 (bs, 1H), 5.02 (tt, J = 4.4 Hz, 7.2 Hz, (t, J = 2.8 Hz, 13.2 Hz, 2H), 2.26 (qd, J = 4.8 Hz, 11.6 Hz, 2H), 2.17 (s,

Example 20. (±) -1- (3- ( 4- amino-3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1- Yl) piperidin-1-yl) prop-2-en-1-one (Compound No. 20)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.67 (bs, 1H), 8.36 (s, 1H), 7.88-7.86 (m, 2H), 7.54-7.50 (m, 3H), 7.16 (s, 1H), (M, IH), 5.93 (m, IH), 4.93-4.85 (m, ), 4.19-4.02 (m, 1H), 3.84-3.45 (m, 1H), 3.32-3.11 (m,

Example 21. (R) -1- (3- ( 4- amino-3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1- Yl) piperidin-1-yl) prop-2-en-1-one (Compound No. 21)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.67 (bs, 1H), 8.36 (s, 1H), 7.88-7.86 (m, 2H), 7.54-7.50 (m, 3H), 7.16 (s, 1H), (M, IH), 5.93 (m, IH), 4.93-4.85 (m, ), 4.19-4.02 (m, 1H), 3.84-3.45 (m, 1H), 3.32-3.11 (m,

Example 22. 3- (5-cyclopropyl-a-3-yl) -1 H - Synthesis of pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 22)

¹ H NMR (400 MHz, DMSO) [delta] 13.94 (bs, IH), 8.20 (s, IH), 8.06 (bs, IH), 7.99 , ≪ / RTI > 1H), 1.15-1.10 (m, 2H), 1.03-0.99

Example 23 1-cyclohexyl-3- (5-cyclopropyl-a-3-yl) -1 H - Synthesis of pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 23)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.62 (bs, 1H), 8.32 (s, 1H), 6.54 (s, 1H), 5.89 (bs, 1H), 4.78-4.70 (m, 1H), 2.14- 2H), 1.34 (m, 1H), 2.07 (m, 1H), 2.05-1.99 (m, 3H), 1.94-1.91 (m, 2H), 1.08-1.02 (m, 2H), 1.28-1.25 (m,

Example 24 3- (5-cyclopropyl-a-3-yl) -1- (tetrahydro -2 H - Failan-4-yl) -1 H - pyrazolo [3,4- d] pyrimidine -4-amine (Compound No. 24)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.65 (bs, 1H), 8.33 (s, 1H), 6.54 (s, 1H), 5.91 (bs, 1H), 4.99 (tt, J = 4.4 Hz, 11.6 Hz , 1H), 4.15 (dd, J = 4.4 Hz, 11.6 Hz, 2H), 3.63 (td, J = 1.6 Hz, 12.0 Hz, 2H), 2.39 (qd, J = 4.4 Hz, 12.0 Hz, 2H), 2.16 -2.09 (m, 1H), 1.96 (dd, J = 2.4 Hz, 12.8 Hz, 2H), 1.17-1.11 (m, 2H), 1.09-1.04 (m, 2H)

Example 25. Synthesis of 3- (5-cyclopropylisoxazol-3-yl) -1-heptyl- 1H -pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.63 (bs, 1H), 8.33 (s, 1H), 6.52 (s, 1H), 6.10 (bs, 1H), 4.39 (t, J = 7.2 Hz, 2H) , 2.16-2.08 (m, 1H), 1.92 (quin, J = 7.2 Hz, 2H), 1.33-1.24 (m, 8H) (t, J = 6.4 Hz, 3 H)

Example 26. Synthesis of 3- (5-cyclopropylisoxazol-3-yl) -1-methyl- 1H -pyrazolo [3,4- d ] pyrimidin-

^{1 H NMR (400 MHz, CDCl} 3) δ 8.60 (bs, 1H), 8.32 (s, 1H), 6.48 (s, 1H), 6.31 (bs, 1H), 4.02 (s, 3H), 2.14-2.05 ( m, 1 H), 1.14-1.03 (m, 2 H), 1.02-1.01 (m, 2 H)

Example 27. 1-Benzyl-3- (5-cyclopropyl-a-3-yl) -1 H - Synthesis of pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 27)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.64 (bs, 1H), 8.37 (s, 1H), 7.34-7.26 (m, 5H), 6.52 (s, 1H), 5.87 (bs, 1H), 5.60 ( (m, 2H), 2.13-2.07 (m, 1H), 1.16-1.05 (m, 2H), 1.05-1.00

Example 28. 1- (4-Amino-3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-1-yl) -2-methyl Propan-2-ol (Compound No. 28)

^{1 H NMR (400 MHz, MeOD} ) δ 8.33 (s, 1H), 6.73 (s, 1H), 4.45 (s, 2H), 2.29-2.22 (m, 1H), 1.29 (s, 6H), 1.24-1.18 (m, 2 H), 1.08 - 1.05 (m, 2 H)

Example 29. 4- (5-ethyl-a-3-yl) -1-isopropyl -1 H - Synthesis of pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 29)

^{1 H NMR (400 MHz, CDCl} 3) δ 8.64 (bs, 1H), 8.33 (s, 1H), 6.66 (s, 1H), 5.90 (bs, 1H), 5.15 (sep, J = 6.8 Hz, 1H) , 2.85 (q, J = 7.6 Hz, 2H), 1.57 (d, J = 6.8 Hz, 6H), 1.38 (t, J = 7.6 Hz, 3H)

Synthesis of pyrazolo [3,4- d] pyrimidin-4-amine (Compound No. 30) - Example 30 1-Isopropyl-3- (isoxazol-3-yl) -1 H

^{1 H NMR (400 MHz, CDCl} 3) δ 8.55 (bs, 1H), 8.47 (d, J = 1.6 Hz, 1H), 8.35 (s, 1H), 7.03 (d, J = 1.6 Hz, 1H), 6.01 (bs, 1H), 5.17 (sep, J = 6.8 Hz, 1H), 1.58 (d, J = 6.8 Hz,

Meanwhile, the novel compounds represented by Formula 1 according to the present invention can be formulated into various forms according to the purpose. The following is a description of some formulations containing the compound of Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

[Formulation example] Method for preparing pharmaceutical agent

Formulation Example 1 Tablets (direct pressurization)

After 5.0 mg of the active ingredient was sieved, 14.1 mg of lactose, 0.8 mg of crospovidone USNF and 0.1 mg of magnesium stearate were mixed and pressurized to make tablets.

Formulation Example 2. Tablet (wet assembly)

After 5.0 mg of the active ingredient was sieved, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of Polysorbate 80 was dissolved in pure water, and an appropriate amount of this solution was added, followed by atomization. After drying, the granules were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressurized to make tablets.

Formulation Example 3. Powder and Capsule

5.0 mg of the active ingredient was sieved and mixed with 14.8 mg of lactose, 10.0 mg of polyvinylpyrrolidone and 0.2 mg of magnesium stearate. The mixture was filtered through a hard No. 5 gelatin capsules.

Formulation Example 4. Injection

100 mg as an active ingredient, 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ .12H ₂ O and 2974 mg of distilled water were added to prepare an injection.

[Experimental Example] Physiological activity test of compound

Experimental Example 1. Measurement of RET kinase inhibitory activity

The inhibitory activity (% inhibition) of RET (Rearranged during transfaction) kinase was measured for the compound represented by Formula 1 according to the present invention and is shown in Table 1 below.

division RET (uM) division RET (uM) Compound No. 1 <1 Compound No. 16 <1 Compound No. 2 <0.01 Compound No. 17 <1 Compound No. 3 &Lt; 0.1 Compound No. 18 <1 Compound No. 4 <1 Compound No. 19 <1 Compound No. 5 <1 Compound No. 20 <1 Compound No. 6 <1 Compound No. 21 <1 Compound No. 7 &Lt; 0.1 Compound No. 22 &Lt; 0.1 Compound No. 8 &Lt; 0.1 Compound No. 23 &Lt; 0.1 Compound No. 9 <1 Compound No. 24 &Lt; 0.1 Compound No. 10 <1 Compound No. 25 &Lt; 0.1 Compound No. 11 <1 Compound No. 26 &Lt; 0.1 Compound No. 12 <5 Compound No. 27 &Lt; 0.1 Compound No. 13 <1 Compound No. 28 &Lt; 0.1 Compound No. 14 <5 Compound No. 29 &Lt; 0.1 Compound No. 15 <1 Compound No. 30 <1

According to the results shown in Table 1, the compounds of the present invention were found to have excellent RET kinase inhibitory activity, and the compounds of Nos. 2, 3, 7, 8 and 22 to 29 were excellent in inhibitory activity.

Experimental Example 2. Measurement of inhibitory activity against RET mutant species

As a representative compound according to the present invention, the inhibitory activity of RET mutant kinase was evaluated against the compound of Compound No. 2. To evaluate the inhibitory activity of the RET mutant kinase, the inhibitory activity of the biotinylated peptide (EGPWLEEEEEAYGWMDF) was used as an indicator by using RET mutant kinase with GST tag added at the N terminus of the RET kinase domain (Carna Biosciences, Millipore) Respectively. Detection of the biotinylated phosphorylated peptide was carried out by the TR-FRET method using an antioxidant antibody conjugated with a Europium label. The inhibitory activity against each mutant of RET kinase was calculated by IC ₅₀ value and is shown in Table 2 below.

RET IC ₅₀ (nM) Cabozantinib
(Control drug) Compound No. 2 RET (wt) 9 <10 RET (G691S) 14 <10 RET (Y791F) 7 <10 RET (V804L) 230 <30 RET (V804M) 358 <10 RET (S891A) 5 <10 RET (M918T) 18 <10

Experimental Example 3. Comparison of inhibitory activity against thyroid cancer cell line and normal cell line

In this experiment, the thyroid cancer cell line TT (American Type Culture Collection) having the normal thyroid epithelial cell line (Nthy-ori 3-1) and the RET kinase activating mutation (C634W) , And the cell proliferation inhibitory activities of Parental Ba / F3 cell line and RET-Ba / F3 cell line transformed without deformation were compared.

12K Nutrient Mixture (manufactured by Life Technologies Corporation (Japan)) containing 10% FBS so as to have 5,000 cells / well, Nthy-ori 3-1 cells, TT cells, Parental Ba / F3 cells and RET- ) And cultured overnight at 37 ° C in the presence of 5% CO ₂ . The experimental compound was then added to dimethyl sulfoxide, diluted to a final concentration of 1 nM to 10 [mu] M and added to a 96 well plate. After incubation for 5 days in the presence of 5% CO ₂ at 37 ° C, CellTiter-Glo (R) Luminescent Cell Viability Assay was added and stirred, and the luminescence intensity was measured using a luminescence measuring device (Envision) . Table 3 below shows the results of measuring the GI ₅₀ values of each test compound.

division GI ₅₀ (uM) Nthy ori-3-1 TT Parental Ba / F3 RET-Ba / F3 Cabozantinib 4.9 0.115 > 10 0.05 Compound No. 2 > 10 <0.3 > 10 &Lt; 0.1 Compound No. 8 > 10 <5 N.A. N.A. Compound No. 13 > 10 <10 <10 <3 Compound No. 14 > 10 <10 > 10 <10 Compound No. 16 > 10 <5 > 10 <10 Compound No. 19 > 10 <5 N.A. N.A. Compound No. 20 > 10 <10 N.A. N.A. Compound No. 23 > 10 <1 > 10 <1 Compound No. 24 > 2 <1 > 10 <1 Compound No. 26 > 10 <5 > 10 <3 Compound No. 28 <10 <5 > 10 <3 Compound No. 29 > 10 <5 > 10 <3 Compound No. 30 > 10 <10 > 10 <5

According to the results shown in the above Table 3, the compounds of the present invention exhibit an abnormally expressed cell line (TT cell or RET-1 cell line) without showing inhibitory activity against a normal cell line (Nthy-ori 3-1 cell, parental Ba / F3 cell) Ba / F3 cells) of the present invention.

Claims (15)

A pharmaceutical composition comprising 3- (isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin-4-amine compound represented by the following formula 1, a pharmaceutically acceptable salt, a hydrate thereof, Lt; RTI ID = 0.0 &gt; isomers:
[Chemical Formula 1]

In Formula 1,

Represents a single bond or a double bond;
A is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; A C ₆ -C ₁₅ aryl group; Or a five to six-membered heteroaryl group containing 1 to 2 hetero atoms selected from S and N;
R is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; C ₁ ~C ₁₀ hydroxyalkyl group; Benzyl group; Or a five to six-membered saturated or partially saturated heterocycloalkyl group containing from one to two heteroatoms selected from O and N;
n represents an integer of 0 to 4;
Wherein said aryl or heterocycloalkyl group is optionally substituted with at least one substituent selected from the group consisting of halo, C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, R ¹ -C (O) - wherein R ¹ is C ₁ -C ₁₀ alkyl or C ₂ -C ₁₀ alkenyl), or R ² -S (O) ₂ -, wherein R ² is C ₁ -C ₁₀ alkyl.
The method according to claim 1,
A represents a hydrogen atom; A tert -butyl group, a cyclobutyl group, a n-pentyl group, an isopentyl group, a neopentyl group, a tert -butyl group, a tert -butyl group, a tert- An alkyl group selected from a pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A phenyl group substituted or unsubstituted with substituents selected from halo, methyl, ethyl, normal propyl, isopropyl, cyclopropyl, methoxy and ethoxy; A thiophene group; Or a pyridinyl group.
The method according to claim 1,
R is a hydrogen atom; A tert -butyl group, a cyclobutyl group, a n-pentyl group, an isopentyl group, a neopentyl group, a tert -butyl group, a tert -butyl group, a tert- An alkyl group selected from a pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A hydroxyalkyl group selected from a 2-methyl-2-hydroxypropyl group, a 2-hydroxybutyl group and a 4-hydroxybutyl group; Benzyl group; Tetrahydro-2H - pyranyl group; Or a piperidinyl group substituted or unsubstituted with R ¹ -C (O) - or R ² -S (O) ₂ -, wherein R ¹ is a C ₁ -C ₆ alkyl group or a C ₂ -C ₆ alkenyl group , R ² is a compound wherein represents a C ₁ ~C ₆ alkyl group.
The method according to claim 1,
remind

Represents a double bond,
A represents a methyl group, an ethyl group, a normal propyl group, an isopropyl group, a cyclopropyl group, a thiophen-2-yl group or a thiophen-
Wherein R is a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a cyclopropyl group, a cyclopropyl group, a normal butyl group, isobutyl group, tert - butyl group, cyclobutyl group, n-pentyl group, isopentyl group, neopentyl A tert -pentyl group, a cyclopentyl group, a normal hexyl group, an isohexyl group, a cyclohexyl group and a normal heptyl group; A hydroxyalkyl group selected from a 2-methyl-2-hydroxypropyl group, a 2-hydroxybutyl group and a 4-hydroxybutyl group; Benzyl group; Or a tetrahydro-2H - pyranyl group,
Wherein n represents 0.
The method according to claim 1,
1-Isopropyl-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
L-Isopropyl-3- (5-cyclopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5-isopropylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5- ( tert -Butyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5-Cyclopentylisoxazol-3-yl) -l-isopropyl-lH - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5-Cyclohexylisoxazol-3-yl) -l-isopropyl-lH - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (thiophen-3-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (thiophen-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5- (4-Fluorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (4-methoxyphenyl) isoxazol-3-yl) -1 H- pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5- (3-Chlorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5- (2-Chlorophenyl) isoxazol-3-yl) -1-isopropyl-1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (pyridin-2-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (pyridin-3-yl) isoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5-Benzylisoxazol-3-yl) -l-isopropyl-lH - pyrazolo [3,4- d ] pyrimidin-4-amine;
1-Isopropyl-3- (5- (phenyl-4,5-dihydroxyisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5-Phenylisoxazol-3-yl) -1- (tetrahydro-2H-pyran - 4-yl) -1H - pyrazolo [3,4- d ] pyrimidin-4-amine;
1- (1- (methylsulfonyl) piperidin-4-yl) -3- (5- (phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin- 4-amine;
1- (4- (4-amino-3- (5- (phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1-yl) piperidin-yl ) Ethanone;
1- (3- (4-amino-3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1-yl) piperidin-1 Yl) prop-2-en-1-one;
(±) -1- (3- (4- amino-3- (5-phenyl-3-yl) -1 H- pyrazolo [3,4 -d] pyrimidin-1-yl) piperidine 1-yl) prop-2-en-1-one;
(4-amino-3- (5-phenylisoxazol-3-yl) -1H - pyrazolo [3,4- d ] pyrimidin- 1-yl) prop-2-en-1-one;
3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine;
1-cyclohexyl-3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine;
3- (5-cyclopropyl-a-3-yl) -1- (tetrahydro -2 H - Failan-4-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine ;
3- (5-Cyclopropylisoxazol-3-yl) -1-heptyl-1 H -pyrazolo [3,4- d ] pyrimidin-4-amine;
3- (5-Cyclopropylisoxazol-3-yl) -1-methyl- 1H -pyrazolo [3,4- d ] pyrimidin-4-amine;
L -Benzyl-3- (5-cyclopropylisoxazol-3-yl) -1H-pyrazolo [3,4- d ] pyrimidin-4-amine;
1- (4-Amino-3- (5-cyclopropyl-a-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-1-yl) -2-methyl-propan-2 Come;
3- (5-ethyl-a-3-yl) -1-isopropyl -1 H - pyrazolo [3,4- d] pyrimidin-4-amine;
1-isopropyl-3- (isoxazol-3-yl) -1 H - pyrazolo [3,4- d] pyrimidin-4-amine.
A pharmaceutical composition for the prophylaxis and treatment of thyroid cancer, lung cancer or breast cancer, which comprises the compound of any one of claims 1 to 5 as an active ingredient.
A therapeutic agent for thyroid cancer, which comprises the compound of any one of claims 1 to 5 as an active ingredient.
A therapeutic agent for lung cancer comprising the compound of any one of claims 1 to 5 as an active ingredient.
(I) reacting a compound represented by the following formula (2) with a compound represented by RX to prepare a compound represented by the following formula (3) into which various Rs have been introduced;

(Wherein R is as defined in claim 1 and X represents a halogen atom)
Ii) a step of preparing a compound represented by the following general formula (IV) in which a vinyl group is introduced by a Stille reaction of a compound represented by the following general formula (3);

(In the above scheme, R is as defined in claim 1)
Iii) converting the vinyl group of the compound represented by the following formula (4) into an aldehyde group by oxdative cleavage and then converting it into an oxime group by reaction with hydroxylamine (NH ₂ OH) A process for producing a compound represented by the formula (5);

(In the above scheme, R is as defined in claim 1)
(Iv) preparing a compound represented by the following formula (1) by a nitrile oxide cycloaddition reaction of a compound represented by the following formula (5);

(In the above scheme,

, A, R and n are each as defined in claim 1,
(Isoxazol-3-yl) -pyrazolo [3,4- d ] pyrimidin-4-amine.
10. The method of claim 9,
Wherein the i) reaction is carried out in the presence of a base of an alkaline earth metal carbonate at 70 캜 to 80 캜.
10. The method of claim 9,
(Ii) the Stille reaction is carried out under reflux at a temperature of 120 ° C to 150 ° C using a metal catalyst of palladium (Pd) or nickel (Ni) and a coupling reagent for an organotin compound Gt;
10. The method of claim 9,
(Iii) the oxidative cleavage reaction is carried out in the presence of an osmium tetroxide (OsO ₄ ) catalyst and an N -methylmorpholine- N -oxide oxidant, followed by the addition of sodium periodate (NaIO ₄ ) &Lt; / RTI &gt;
10. The method of claim 9,
Wherein the iv) cyclization addition reaction is carried out under a condition using a hyperbalanced iodine compound selected from (diacetoxy iodo) benzene or phenyl iodine bis (trifluoroacetate).
An intermediate compound represented by the following formula (4):
[Chemical Formula 4]

In Formula 4,
R is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; C ₁ ~C ₁₀ hydroxyalkyl group; Benzyl group; Or a five to six-membered saturated or partially saturated heterocycloalkyl group containing from one to two heteroatoms selected from O and N;
An intermediate compound represented by the following formula (5):
[Chemical Formula 5]

In Formula 5,
R is a hydrogen atom; A C _{1 to} C ₁₀ alkyl group; C ₁ ~C ₁₀ hydroxyalkyl group; Benzyl group; Or a five to six-membered saturated or partially saturated heterocycloalkyl group containing from one to two heteroatoms selected from O and N;