KR100368515B1 - Cyclin dependent kinase inhibitor and method of preparing the same - Google Patents

Abstract

The present invention relates to a cyclin dependent kinase inhibitor having the formula (1). This compound has a very good effect of inhibiting cyclin dependent kinase activity and has anticancer activity, and thus can be usefully used as an active ingredient of an anticancer agent composition.

[Formula 1]

(Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, and R ₃ represents an alkyl group.)

Description

Cyclin-dependent kinase inhibitors and methods for their preparation {CYCLIN DEPENDENT KINASE INHIBITOR AND METHOD OF PREPARING THE SAME}

The present invention relates to a cyclin-dependent kinase inhibitor and a method for producing the same, and more particularly, to a cyclin-dependent kinase inhibitor of the formula (1) having excellent cyclin-dependent kinase inhibitory activity.

[Formula 1]

(Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, and R ₃ represents an alkyl group.)

It was in the late 1980s that he began to actively study cell division processes at the molecular level. In the late 1980s, research began to become active through the study of the division of frog eggs, the characterization of various cell growths and radioactive mutations in yeast, and the study of the Rb gene, a tumor suppressor. The mechanism of cell division regulation, which began to be revealed in more detail in the 90's, revealed that small cell growth regulators regulate cell growth, differentiation, development, aging and apoptosis through cell growth regulation.

These findings have helped to better understand the pathological phenomena of many diseases, and cancer is a prime example. In the process of transforming normal cells into cancer cells, many cell growth regulators have lost their function. As a result of analyzing cancer cells, the activity of cell growth regulators was often different from that of normal cells. In particular, the difference in the activity of cell growth regulators between cancer cells and normal cells was found to correlate with the invasion and metastasis of cancer, which is the most problematic in cancer pathology. In addition, cell cycle deregulation is induced as a result of overexpression of elements that regulate cell growth or knock-out that inactivates the gene of interest using transgenic animals. ) Is a direct cause of cancer.

The cell growth process, like all other biological controls, is subject to positive and negative regulation. The main framework of cell cycle regulation known to date is determined by cycline dependent kinase (hereinafter referred to as "CDK") activity, and this kinase activity is subject to positive or minor control depending on the environment in which the cell is located. . Many cancer cells and studies of carcinogenic mechanisms have been found to cause problems in the control or secondary regulation. In other words, excessive over- or under-regulation and the timely regulation, which is an important aspect of cell growth regulation, have been pointed out as problems in cancer cells.

Three representative cyclin dependent kinases in mammals are listed. The first is CDK4, which is active in the mid-G1 phase of the cell cycle, the second is CDK2, which is active in the middle and S phases of G1, and CDK1, which is active in the G2-M phases. It is known that CDK4 and CDK2 are regulated by G1-S cell cycle checkpoint and CDK1 is regulated by G2-M checkpoint. In many cancer cells, the regulatory mechanisms of CDK4, CDK2 and CDK1 have been shown to be abnormal, and in fact, the artificially induced non-ideality of these enzymes in transgenic animals has been shown to cause cancer. This suggests that, among other cyclin dependent kinases, CDK4, CDK2 and CDK1 are the most promising targets as anticancer agents.

First, the case of CDK4 is described in more detail. The association between non-ideal regulation of CDK4 activity and cancer induction is well demonstrated in various cancer tissues. Deletion of the p16 and p15 genes has been reported in several cancers, and overexpression of cyclin D1 in particular has been shown in several cancers, particularly in the context of breast cancer metastasis. This suggests that deregulated CDK4 activity may be a factor in the malignant phenotype of cancer cells. It has been reported that p16 gene knockout mice develop cancer as well as p53 gene knockout mice, suggesting that the loss of CDK4 function in the p16 gene is responsible for cancer. In addition, in the NIH 3T3 cells overexpressing the Ras gene or the src gene, CDK4 is further activated downstream, thereby showing a possibility to play a role. Conversely, the modified phenotype transformed from the p16 gene or the p21 gene with the ras gene was observed to change to the normal phenotype.

The experimental evidence listed above demonstrates that it is clear that deregulation of CDK4 activity is responsible for cancer induction, and that it may play a role in maintaining the phenotype of cancer cells. Therefore, it is considered that the inhibitor of CDK4 is highly likely to have an anticancer effect.

In the case of CDK2, overexpression of Cycline E was observed in some breast cancers, and it is well known that it is closely related to the metastasis of breast cancer. Overexpression of Cycline E has been proposed to inhibit cell death in low and low serum conditions and to cause anchorage independent growth. Hyperproliferation (neoplasia) of breast epithelial cells was observed in CDK2 overexpressing transgenic animals using the MMTV promoter. This strongly suggests that CDK2 activity is involved in the cellular transformation process or maintenance thereof, demonstrating the high potential of CDK2 as an anticancer agent.

In addition, it has been found that CDK1, CDK3, CDK5, CDK6, and CDK7 also play important roles at each stage of the cell division process, and these are classified into the same family of cyclin-dependent kinases (CDKs). In addition, in the case of CyClean, CyClean A, B, C, D2, D3, D4, F and CyClean G belong to the same family in addition to CyClean D1 or CyClean E.

Based on these accumulated findings, research has recently begun to develop inhibitors that effectively inhibit these cyclone-dependent kinases (CDKs) as good targets for anticancer drugs.

Representative compounds so far developed as effective CDKs inhibitors are Flavopyridol of Formula A, which is disclosed in European Patent Nos. 0,241,003 (1987) and 0,366,061 (1990).

[Formula A]

In addition, CDKs inhibitors having a purine structure of formula B have recently been disclosed in WO 97/16447.

[Formula B]

It is an object of the present invention to provide a novel purine structured cyclin dependent kinase inhibitor which can effectively inhibit cyclin dependent kinase.

Another object of the present invention is to provide a method for preparing a cyclin dependent kinase inhibitor which can easily prepare a compound having the above-described physical properties.

In order to achieve the above object, the present invention provides a cyclin dependent kinase inhibitor of formula (1), a pharmaceutically acceptable salt, hydrate or solvate thereof.

[Formula 1]

(Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, and R ₃ represents an alkyl group.)

The present invention also provides a method for preparing a cyclin dependent kinase inhibitor of Formula 1, comprising reacting a compound of Formula 4 with R ₂ OH.

[Formula 4]

(Wherein R ₁ and R ₃ are the same as defined in Formula 1 above, X represents F, Cl, Br and I.)

The compound of Formula 4 may be prepared by reacting a compound of Formula 2 with NH ₂ R ₁ to prepare a compound of Formula 3, and reacting the compound of Formula 3 with R ₃ X.

[Formula 2]

[Formula 3]

Wherein R ₁ and X are the same as defined in Formula 4 above

In the present invention, the compound of Formula 1 may also be prepared by reacting a compound of Formula 5 with R ₃ X.

[Formula 5]

(Wherein R ₁ and R ₂ are the same as defined in Formula 1 above)

The compound of Formula 5 may be prepared by reacting the compound of Formula 2 with NH ₂ R ₁ at 80 to 100 ° C. in the presence of a base and a solvent, and then reacting the product at a boiling point temperature of 120 ° C. to a solvent.

Hereinafter, the present invention will be described in more detail.

The present invention relates to compounds that inhibit cyclin dependent kinase enzymes. Cycline dependent kinase enzymes in the present invention include CDK2, CDK4 and CDK1, CDK3, CDK5, CDK6 and CDK7, all of which are collectively referred to as CDKs enzymes. In addition, cyclins include cyclins D1 and cyclins E and cyclins A, B, C, D2, D3, D4, F, G.

Inhibitors of CDKs enzymes known in the art, in particular, purine-based compounds, all have a structure in which position 2 is connected to a nitrogen atom, whereas the compound of formula 1 of the present invention has a completely new structure in which position 2 is replaced with an oxygen atom. Compound. This compound has the effect of inhibiting CDKs enzymes very effectively.

[Formula 1]

Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, and in particular, R ₂ represents a C ₂ to C ₆ substituted with 1-2 hydroxyl groups An alkyl group or a C _{2 to} C ₆ alkyl group, and R ₃ represents an alkyl group.)

Representative compounds of the compounds having Formula 1 of the present invention include 1- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -butanol; 1- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -ethan-2-ol; 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1,2-propanediol; 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1-propanol; 2-{[6- (3-chloroaninilino) -9-isopropyl-9H-purin-2-yl] oxy-1-ethanol; 3-{[6- (3-chloroaninilino) -9-isopropyl-9H-purin-2-yl] oxy} -1-propanol; 2- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1,2-propanediol; N- (2-butoxy-9-isopropyl-9H-purin-6-yl) -N- (3-nitrophenyl) amine; N- (2-butoxy-9-isopropyl-9H-purin-6-yl) -1,3-benzenediamine.

The compounds of formula 1 according to the invention may also comprise pharmaceutically acceptable salts, hydrates and solvates. Such pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions, such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, tartaric acid, formic acid, Organic carbonic acids such as citric acid, acetic acid, trichloroacetic acid or trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, and the like Acid addition salts formed by phonic acid or the like.

On the other hand, the compounds according to the invention may have an asymmetric carbon center and therefore may exist as racemic compounds, diastereomeric mixtures and individual diastereoisomers, all of these isomeric forms being included in the scope of the invention.

According to the present invention, the compound of Formula 1 as defined above may be prepared by the following Scheme 1.

Scheme 1

Hereinafter, the reaction scheme 1 will be described in more detail.

The compound of Formula 2 is reacted with NH ₂ R ₁ at a ratio of 1: 1 to 1.5 equivalents in an organic solvent at about 80 ° C. for 3 to 24 hours to prepare the compound of Formula 3. As the organic solvent, any organic solvent generally used in an amine substitution reaction may be used, and representative examples thereof may include alcohols such as n-butanol. As the base, any acid can be used as long as it can neutralize the acid generated in this step, and triethylamine or the like can be used as a representative example.

The obtained compound of formula 3 is reacted with R ₃ X in an organic solvent in the presence of a base at about 90 ° C. for 1 to 20 hours to prepare a compound of formula 4. As the base, any base generally used to prepare an amine compound may be used, and representative examples thereof may include sodium hydride (NaH) and soamide (NaNH ₂ ). In the present invention, as the halogenated compound, any R ₃ X halogen compound in which X is selected from the group consisting of F, Br, Cl, and I may be used. As a representative example thereof, 2-bromopropane may be used. In addition, any organic solvent generally used may be used as the organic solvent, and representative examples thereof may include dimethyl formamide, tetrahydrofuran, CH ₂ Cl ₂ , and the like.

Subsequently, the compound of formula 4 is reacted by heating the compound of formula 4 obtained in the presence of 1 to 1.5 equivalents of alkoxide and R ₂ OH, which is a solvent and a reagent, to a boiling point temperature of 50 ° C. to prepare a compound of formula 1.

Alternatively, the compound of Formula 1 as defined above may be prepared according to Scheme 2 below.

Scheme 2

The compound of Chemical Formula 2 and NH ₂ R ₁ are reacted at a temperature of 80 to 100 ° C. in a presence of a base in an organic solvent of R ₂ OH at a ratio of 1: 1 to 1.5 equivalent for 3 to 5 hours. Subsequently, the product is reacted again for 1 to 3 hours at a boiling point temperature of 120 to the solvent to prepare a compound of Formula 5. When reacting at 80 to 100 ° C., NH ₂ R ₁ participates in the reaction, and when the temperature is raised to 120 to the boiling point of the solvent, the solvent participates in the reaction. When reacting at a high temperature at one time, NH ₂ R ₁ and R ₂ OH may simultaneously participate in the reaction, and R ₁ and R ₂ may be substituted at other positions of the compound of Formula 2, thereby forming an unwanted side reaction. . Therefore, in this way, after the first reaction at a low temperature, the temperature must be raised to the second reaction.

As the organic solvent, any organic solvent generally used in an amine substitution reaction may be used, and representative examples thereof may include alcohols such as n-butanol. As the base, any acid can be used as long as it can neutralize the acid generated in this step, and triethylamine or the like can be used as a representative example.

The obtained compound of formula 5 is reacted with R ₃ X in an organic solvent at a temperature of about 90 ° C. for 1 to 20 hours to prepare a compound of formula 1. In the present invention, as the halogenated compound, any R ₃ X halogen compound in which X is selected from the group consisting of F, Br, Cl, and I may be used. As a representative example thereof, 2-bromopropane may be used. In addition, any organic solvent generally used may be used as the organic solvent, and representative examples thereof may include dimethyl formamide, tetrahydrofuran, CH ₂ Cl ₂ , and the like.

The obtained product may further be subjected to a reduction reaction using a metal catalyst in the presence of hydrogen gas. As the catalyst, any metal catalyst generally used in a reduction reaction may be used, and representative examples thereof may include palladium, nickel, platinum, and the like mixed with carbon.

After the completion of the reaction, the product can be separated and purified by conventional workup methods such as chromatography, recrystallization and the like.

Hereinafter, preferred embodiments of the present invention will be described. However, the following examples are only preferred embodiments of the present invention and the present invention is not limited to the following examples.

Preparation Example 1 Synthesis of N- (2-chloro-9H-purin-6-yl) -N- (4-methoxy-benzyl) amine

1 g (5.29 mmol) of 2,6-dichloropurine and 0.87 g of 4-methoxybenzylamine were added to 30 ml of normal butanol, followed by stirring at 90 ° C. for 15 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, diluted with 50 ml of ethyl acetate, and washed twice with 10 ml of water. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated. The obtained solid was added with a small amount of ethyl acetate and stirred, followed by filtration to obtain 1.4 g (yield 90%) of the white target compound.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.67 (1H, s), 7.17 (2H, d, J = 8.25 Hz), 6.72 (2H, d, J = 8.25 Hz), 4.55 (2H, bs), 3.64 (3H, s)

FAB MS (m / e) = 290 [M ⁺ + 1]

Preparation Example 2 Synthesis of N- (2-Chloro-9-isopropyl-9H-purin-6-yl) -N- (4-methoxy-benzyl) amine

0.5 g (1.7 mmol) of the compound obtained in Preparation Example 1 was dissolved in 20 ml of dimethyl formamide, and 0.25 g of 2-bromopropane was added thereto. 0.1 g of sodium hydride (NaH) was added thereto at room temperature and stirred at 80 ° C. for 17 hours. The reaction was completed, concentrated under reduced pressure, and diluted with 50 ml of ethyl acetate. The organic solution was washed with 5 ml of 0.1 N aqueous hydrochloric acid solution and 20 ml of water and dried over anhydrous magnesium sulfate. After filtration, a small amount of ethyl acetate was added to the solid obtained by concentrating. The mixture was stirred well and filtered to obtain 0.47 g (yield 83%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.73 (1H, s), 7.26 (2H, d, J = 9.15 Hz), 6.81 (2H, d, J = 9.15 Hz), 4.76 (1H, m), 4.66 (2H, bs), 3.73 (3H, s ), 1.50 (6H, d, J = 8.9 Hz)

FAB MS (m / e) = 322 [M ⁺ + 1]

Example 1 Synthesis of 1- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -butanol

0.3 g (0.9 mmol) of the compound prepared in Preparation Example 2 was added to 10 ml of 1,2-dihydroxy butanol, and 86 mg of sodium thibutoxide was added thereto and stirred at 100 ° C. After the reaction was completed, the mixture was neutralized with diluted aqueous hydrochloric acid solution, and water and ethyl acetate were added and the layers were separated. The aqueous layer was extracted with ethyl acetate, concentrated under reduced pressure and purified by chromatography to give 0.21 g (yield 65%) of the desired product.

¹ H NMR (CDCl ₃ , ppm); 7.56 (1H, s), 7.23 (2H, d), 6.81 (2H, d), 4.65 (3H, m), 4.47 (2H, t), 1.7-1.5 (4H, m), 1.6 (6H, d) , 1.0 (3H, t)

FAB MS (m / e) = 370 [M ⁺ + 1]

Example 2 Synthesis of 1- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -ethan-2-ol

144 mg of the target compound was prepared in the same manner as in Example 1, except that 0.3 g (0.9 mmol) of the compound prepared in the Preparation Example 2 was used, except that ethylene glycol was used instead of 1,2-dihydroxy butanol. (Yield 45%) was obtained.

¹ H NMR (CDCl ₃ , ppm); 7.55 (1H, s), 7.21 (2H, d), 6.81 (2H, d), 4.64 (3H, m), 4.45 (2H, m), 3.91 (2H, m), 3.74 (3H, s), 1.49 (6H, d)

FAB MS (m / e) = 358 [M ⁺ + 1]

Example 3 Synthesis of 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1,2-propanediol

Except for using the reagent glycerol instead of 1,2-dihydroxy butanol, using the compound 0.1 g (0.3 mmol) prepared in Preparation Example 2 was carried out in the same manner as in Example 1, 37 mg (yield) 32%).

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.53 (1H, s), 7.21 (2H, d), 6.81 (2H, d), 4.63 (3H, m), 4.47 (2H, m), 3.92 (2H, m), 3.74 (3H, s), 3.29 (1H, m), 1.50 (6H, d)

FAB MS (m / e) = 388 [M ⁺ + 1]

Example 4 Synthesis of 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1-propanol

Except for using the reagent 1,3-propanediol instead of 1,2-dihydroxy butanol was carried out in the same manner as in Example 1 using 0.1 g (0.3 mmol) of the compound prepared in the process of Preparation Example 2 42 mg (yield 38%) of the title compound were obtained.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.54 (1H, s), 7.22 (2H, d), 6.81 (2H, d), 4.65 (3H, m), 4.43 (2H, m), 3.92 (2H, m), 3.74 (3H, s), 1.49 (6H, d), 1-1.6 (2H, m)

FAB MS (m / e) = 372 [M ⁺ + 1]

Preparation Example 3 Synthesis of (2-Chloro-9H-purin-6-yl)-(3-chloro-phenyl) -amine

0.8 g of 3-chloroaniline was used instead of 4-methoxybenzylamine to carry out the same procedure as in Preparation Example 1 to obtain 1 g of the target compound (yield 70%).

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.85 (1H, s), 7.84 (1H, s), 7.59 (1H, d, J = 7.8 Hz), 7.20 (1H, m), 6.99 (1H, d, J = 7.75)

FAB MS (m / e) = 280 [M ⁺ + 1]

Preparation Example 2 Synthesis of 2-chloro-N- (3-chlorophenyl) -9-isopropyl-9H-purin-6-amine

0.45 g (1.7 mmol) of the compound of Preparation Example 3 was used in the same manner as in Preparation Example 2 to obtain 0.35 g (yield 63%) of the title compound.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.89 (2H, m), 7.68 (1H, m), 7.30 (1H, m), 7.10 (1H, m), 4.88 (1H, m), 1.63 (6H, d)

FAB MS (m / e) = 322 [M ⁺ + 1]

Example 5 Synthesis of 2-{[6- (3-chloroanilinino) -9-isopropyl-9H-purin-2-yl] oxy]}-1-ethanol

45 mg (yield 42%) of the title compound was obtained in the same manner as in Example 1, except that 0.1 g (0.31 mmol) of the compound of Preparation Example 4 and ethylene glycol were used.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.91 (2H, m), 7.73 (1H, s), 7.43 (1H, m), 7.21 (1H, m), 4.75 (1H, m), 4.43 (2H, m), 3.89 (2H, m), 1.47 (6H, d)

FAB MS (m / e) = 348 [M ⁺ + 1]

Example 6 Synthesis of 3-{(6- (3-chloroaninynino) -9-isopropyl-9H-purin-2-yl] oxy-1-propanol

Except for using 1,3-propanediol instead of ethylene glycol, using the compound of Preparation Example 4 0.1 g (0.31 mmol) in the same manner as in Example 5 to obtain 37 mg (yield 33%) of the target compound. Got it.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 7.91 (2H, m), 7.73 (1H, s), 7.43 (1H, m), 7.21 (1H, m), 4.75 (1H, m), 4.37 (2H, m), 3.76 (2H, m), 1.65 (2H, m), 1.47 (6H, d)

FAB MS (m / e) = 362 [M ⁺ + 1]

Example 7 Synthesis of 2- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -1,2-propanediol

Except that glycerol was used instead of ethylene glycol, it carried out similarly to Example 5, and obtained 32 mg (yield 28%) of the target compounds.

FAB MS (m / e) = 378 [M ⁺ + 1]

Preparation Example 5 Synthesis of N- (2-butoxy-9H-purin-6-yl) -3-nitrophenyl) amine

0.5 g (2.6 mmol) of 2,6-dichloropurine, 0.43 g of 3-nitroaniline and 7 ml of triethylamine were added to 15 ml of normal butanol, followed by stirring at 90 ° C. for 3 hours. The temperature was further raised to reflux for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, diluted with 30 ml of ethyl acetate, and washed twice with 10 ml of water. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated. The obtained solid was added with a small amount of ethyl acetate, and stirred and filtered to obtain 0.72 g (yield 85%) of the title compound, which was white.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm); 9-7.25 (5H, m), 4.53 (2H, t), 2.7-1.62 (4H, m), 1.1 (3H, t)

FAB MS (m / e) = 329 [M ⁺ + 1]

Example 8 Synthesis of N- (2-butoxy-9-isopropyl-9H-purin-6-yl) -N- (3-nitrophenyl) amine

In the same manner as in Preparation Example 2, using 0.1 g (0.3 mmol) of the compound of Preparation Example 5, 87 mg (yield 77%) of the target compound was obtained.

¹ H NMR (CDCl ₃ , ppm); 7.9-7.45 (5H, m), 4.8 (1H, m), 4.5 (2H, t), 1.7-1.5 (4H, m), 1.6 (6H, d), 0.9 (3H, t)

FAB MS (m / e) = 372 [M ⁺ + 1]

Example 9 Synthesis of N- (2-butoxy-9-isopropyl-9H-purin-6-yl)-(3-aminophenyl) amine

10 mg (0.03 mmol) of the compound prepared in Example 8 was dissolved in 5 ml of methanol and reduced under hydrogen atmospheric pressure using a palladium / carbon catalyst to obtain 8.2 mg (yield 90%) of the target compound.

¹ H NMR (CDCl ₃ , ppm); 7.8-7.7 (2H, m), 7.3-7.1 (3H, m), 4.8 (1H, m), 4.5 (2H, t), 1.7-1.5 (4H, m), 1.6 (6H, d), 1.0 ( 3H, t)

FAB MS (m / e) = 341 [M ⁺ + 1]

Experimental Example 1 Inhibitory Activity of CDK2 and CDK4

CDK2 and CDK4 inhibitory activity was measured using the compounds prepared by the methods of Examples 1 to 9. The assay for the inhibition of CDK2 protein was performed using the Kitagawa method (Kitagawa, M. et al., Oncogene, 9: 2549, 1994), and the Carlson method for CDK4 (Carlson, BA et al .; Cancer Research 56: 2473, 1996).

As CDK2, insect cell extracts or active enzymes purified therefrom which simultaneously infected baculovirus expressing CDK2 gene and baculovirus expressing cyclin A gene were used. As a CDK4 enzyme, one obtained from an insect cell simultaneously infected with a baculovirus expressing the CDK4 gene and a baculovirus expressing the Cyclin D1 gene was also used. Histone H1 or Rb protein was used as a substrate of CDK2, and Rb protein was used as a substrate of CDK4. Compounds diluted by concentration were added, and an appropriate amount of CDK2 / cyclin A or CDK4 / cyclin D1 was added to the substrate protein, and [gamma-32P labeled] ATP, and then the substrates were separated to measure radioactivity contained in the substrate.

According to the method described above, the inhibitory ability of each of the inhibitors according to the present invention measured for CDK2 and CDK4 is shown as an IC 50 value. The results are as shown in Table 1 below.

rescue constitutional formula FAB MS (M + H) CDK2IC50 (μM) CDK4IC50 (μM) Example 1 C ₂₀ H ₂₇ N ₅ O ₂ 370 <5 μM <50 μM Example 2 C ₁₈ H ₂₃ N ₅ O ₃ 358 <1 μM <50 μM Example 3 C ₁₉ H ₂₃ N ₅ O ₄ 388 <1 μM <50 μM Example 4 C ₁₉ H ₂₅ N ₅ O ₃ 372 <1 μM <50 μM Example 5 C ₁₆ H ₁₈ ClN ₅ O ₂ 348 <2 μM <250 μM Example 6 C ₁₇ H ₂₀ ClN ₅ O ₂ 362 <5 μM <50 μM Example 7 C ₁₇ H ₂₀ ClN ₅ O ₂ 371 <3 μM <50 μM Example 8 C ₁₈ H ₂₂ N ₆ O ₃ 371 <3 μM <50 μM Example 9 C ₁₈ H ₂₄ N ₆ O 341 <2 μM <50 μM

As shown in Table 1, it can be seen that the compound of the present invention has an excellent effect of inhibiting cyclin dependent kinase activity.

As described above, the compound of the present invention is a purine-based compound in which position 2 is substituted with an oxygen atom, and is very excellent in inhibiting cyclin-dependent kinase activity. Therefore, since such compounds have anticancer activity, they can be usefully used as active ingredients of anticancer agent compositions.

Claims (7)

Cycline dependent kinase inhibitors represented by Formula 1, pharmaceutically acceptable salts, hydrates or solvates thereof: [Formula 1] Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, and R ₃ represents an alkyl group. The method of claim 1, Said R ₂ is a C ₂ -C ₆ alkyl group or C ₂ -C ₆ alkyl group substituted with ₁ to 2 hydroxyl groups, a cyclin dependent kinase inhibitor, a pharmaceutically acceptable salt, hydrate or solvate thereof. The method of claim 1, Inhibitor of the formula 1 is 1- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy] -butanol, 1- [9-isopropyl-6- (4 -Methoxy-benzylamino) -9H-purin-2-yloxy] -ethan-2-ol, 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2- Yloxy] -1,2-propanediol, 3- [9-isopropyl-6- (4-methoxy-benzylamino) -9H-purin-2-yloxy-1-propanol, 2- {6- ( 3-chloroaninilino) -9-isopropyl-9H-purin-1-2-yl] oxy-1-ethanol, 3-{[6-3-chloroaninilino) -9-isopropyl-9H-purine- 2-yl] oxy} -1-propanol, 3-{[6-chloroanilinino) -9-isopropyl-9H-purin-2-yl] oxy} -1,2-propanediol, N- (2- Butoxy-9-isopropyl-9H-purin-6-yl) -N- (3-nitrophenyl} amine, N- (2-butoxy-9-isopropyl-9H-purin-6-yl)- Cycline kinase inhibitor, a pharmaceutically acceptable salt, hydrate or solvate thereof, which is a compound selected from the group consisting of 1,3-benzenediamine. Reacting the compound of Formula 2 with R ₂ OH Method for producing a cyclin-dependent kinase inhibitor represented by the following formula (1) comprising a. [Formula 1] [Formula 2] Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, R ₃ represents an alkyl group, and X represents F, Cl, Br and I. The method of claim 4, wherein Compound of Formula 2 is prepared by reacting a compound of Formula 3 with NH ₂ R ₁ to prepare a compound of Formula 4; A process for preparing a compound of Formula 4, which is prepared by reacting a compound of Formula 4 with R ₃ X [Formula 2] [Formula 3] [Formula 4] Wherein R ₁ represents a substituted or unsubstituted benzyl group or an aryl group, R ₂ represents a substituted or unsubstituted alkyl group, R ₃ represents an alkyl group, and X represents F, Cl, Br and I. Reacting a compound of formula 5 with R ₃ X Method for producing a cyclin-dependent kinase inhibitor represented by the following formula (1) comprising: [Formula 1] [Formula 5] Wherein R ₁ represents a substituted or unsubstituted benzyl or aryl group, R ₂ represents a substituted or unsubstituted alkyl group, R ₃ represents an alkyl group, and X represents F, Cl, Br and I Indicates. The method of claim 6, The compound of Formula 5 is formed by reacting a compound of Formula 2 with NH ₂ R _{1 at} 80 to 100 ° C. in an organic solvent, and then reacting the product at a boiling point temperature of the solvent of the compound of Formula 2 to 120 ° C. The preparation method which is a compound: [Formula 2] [Formula 5] Wherein R ₁ represents a substituted or unsubstituted benzyl or aryl group, R ₂ represents a substituted or unsubstituted alkyl group, R ₃ represents an alkyl group, and X represents F, Cl, Br and I Indicates.