KR100392468B1 - 3-Hydroxychromene-4-on derivatives useful as cyclin dependendent kinase inhibitors - Google Patents

Abstract

The present invention relates to 3-hydroxychromen-4-one derivatives of the general formulas (1), (2) or (3) below, which are useful as inhibitors of cyclin dependent kinases (CDKs), pharmaceutically acceptable salts, hydrates, solvates and isomers thereof. . The present invention also provides a method for preparing a compound of formula 1, 2 or 3 and cancer, inflammation, vascular stenosis, characterized in that it contains a compound of formula 1, 2 or 3 as an active ingredient together with a pharmaceutically acceptable carrier and The present invention relates to inhibitors and therapeutic agents for diseases related to cell proliferation such as angiogenesis.

[Formula 1]

[Formula 2]

[Formula 3]

In the above formula,

R 1 is hydrogen, alkyl, amino alkyl or hydroxy alkyl group,

R 2 is hydrogen, methyl, trifluoromethyl or halogen,

R3 represents an alkyl, amino alkyl or hydroxy alkyl group,

X represents a halogen.

Description

3-Hydroxychromene-4-on derivatives useful as cyclin dependendent kinase inhibitors}

The present invention provides novel 3-hydroxychromen-4-one derivatives of the general formulas (1), (2) or (3) below, useful as inhibitors of cyclin dependent kinase (hereinafter referred to as CDK), pharmaceutically acceptable salts thereof, Hydrates, solvates and isomers, methods for producing the same, and anticancer drug compositions excellent in antiproliferative effects containing them as active ingredients.

[Formula 1]

[Formula 2]

[Formula 3]

In the above formula,

R 1 is hydrogen, alkyl, amino alkyl or hydroxy alkyl group,

R 2 is hydrogen, methyl, trifluoromethyl or halogen,

R3 represents an alkyl, amino alkyl or hydroxy alkyl group,

X represents a halogen.

The full-scale study at the molecular level of the cell division process began to be active in the late 1980s through the study of the division of frog eggs, the growth of various cells in yeast, the characterization of radioactive mutations, and the study of tumor suppressor Rb. According to the mechanism of cell division regulation, which became more detailed in the 90s, it was found that small cell growth regulators centrally regulate cell growth, differentiation, development, aging, and apoptosis through cell growth regulation. Turned out. These findings have already helped to better understand the pathological phenomena of many diseases. A representative example is cancer. In the process of transforming normal cells into cancer cells, many cell growth regulators have lost their function. According to the analysis of cancer cells, the activity of cell growth regulators is often different from that of normal cells, especially when it shows a deep correlation with cancer invasion and metastasis, which is the most problematic in cancer pathology. have. Experimental results of overexpression or knock-out of elements that regulate cell growth using transgenic animals suggest that cancer in these animals can cause cell cycle deregulation. deregulation has been shown to be a direct cause of cancer.

The cell growth process, like all other biological controls, is subject to positive and negative regulation. To date, studies have shown that the cell growth cycle is largely regulated by the activity of cyclin dependent kinases, and many cancer cells or carcinogenesis mechanisms have been associated with problems of positive or negative regulation of kinase activity. In other words, cancer may develop if there is a lack of balanced regulation or timely regulation.

Three representative cyclin dependent kinases in mammals are CDK4 (cyclin dependent kinase 4) and CDK2, which are active in the G1-S phase of the cell cycle, and CDC2 (CDK1), which are active in the G2-M phase. It is known that CDK4 and CDK2 are regulated by G1-S cell cycle checkpoint and CDC2 is regulated by G2-M checkpoint. Regulatory mechanisms of CDK4 and CDK2 and CDC2 (CDK1) have been shown to be abnormal in several cancer cells, and in fact, the artificially induced abnormalities of these enzymes have been shown to cause cancer in experiments with transgenic animals. . Thus, these representative cyclin dependent kinases CDK4, CDK2 and CDC2 (CDK1) are promising targets for anticancer drug development. These kinases are also targeted for the development of inhibitors and therapeutic agents for diseases related to cell proliferation such as inflammation, vascular stenosis and angiogenesis.

The results reported so far regarding the association between these CDKs and cancer disease are described in more detail below.

The association between abnormal regulation of CDK4 activity and cancer induction has been observed in several cancer tissues. Deletion of the p16 and p15 genes or the overexpression of cyclin D1, which is essential for the activity of CDK4, has been identified in a number of cancers, resulting in a malignant phenotype of cancer cells when CDK4 activity is not regulated. phenotype) may be present. In addition, the incidence of cancer in p16 knockout mice is as high as that of p53 knockout mice, suggesting that loss of CDK4 function by p16 causes cancer. These experimental evidences seem to prove that deregulation of CDK4 activity is a clear cause of cancer, and furthermore, it is likely to play a role in maintaining the phenotype of cancer cells. Therefore, CDK4 inhibitors are very likely to have anticancer effects.

On the other hand, in the case of CDK2, overexpression of cyclin E, which is essential for the activity of CDK2 in some breast cancers, has been observed and it is well known that it is deeply associated with metastasis of breast cancer. Overexpression of cyclin E has been shown to inhibit cell death in low serum conditions and to cause anchorage independent growth. Hyperproliferation and neoplasia of mammary epithelial cells were observed in CDK2 overexpressing transgenic animals using the MMTV promoter. This strongly suggests that CDK2 activity is involved in the process of cell transformation or its maintenance, indicating that inhibitors of CDK2 can act as anticancer agents.

In addition, CDC2 (CDK1), CDK3, CDK5, CDK6, CDK7, etc. are gradually found to play an important role in each stage of cell division, and are divided into families of the same cyclin-dependent kinase (CDKs). In the case of cyclin, cyclins A, B, C, D2, D3, D4, F, and G belong to the same family in addition to cyclin D1 or cyclin E mentioned above.

Based on these accumulated findings, the recurrence of these inhibitors has recently begun to occur, recognizing that inhibitors that effectively inhibit these cyclin dependent kinases (CDKs) will be useful as anticancer agents.

Compounds effective as CDKs inhibitors developed so far include the flavopyridols of Formula A (see European Patent Nos. 0,241,003 (1987) and 0,366,061 (1990)).

[Formula A]

Next, CDKs inhibitors having a Purine structure of Formula B have been disclosed in WO 97/20842.

[Formula B]

In addition, CDKs inhibitors having the 4-aminopyrimidine structure of Formula C have recently been disclosed in WO 98/33798.

[Formula C]

However, the CDK inhibitors developed to date have not yet had a satisfactory effect. Therefore, the present inventors have conducted intensive studies on inhibitors of these CDKs enzymes, especially flavone-based compounds. It was confirmed that the compounds of 1, 2 or 3 effectively inhibited the above-mentioned CDKs enzymes and completed the present invention.

Accordingly, an object of the present invention is a 3-hydroxychromen-4-one derivative of Formula 1, 2 or 3, a method for preparing the derivative and cancer, inflammation, vascular stenosis and angiogenesis containing the derivative as an active ingredient. The present invention provides an inhibitor and therapeutic composition for diseases related to cell proliferation. CDKs include CDK2, CDK4 and CDC2 (CDK1), CDK3, CDK5, CDK6, CDK7, and the like, and cyclin cyclin D1 and cyclin E and cyclin A, B, C, D2, D3, D4, F, G It includes everything.

Hereinafter, the present invention will be described in detail.

The present invention is a novel 3-hydroxychromen-4-one of formula (1), (2) or (3), which exhibits the inhibitory and therapeutic effect of diseases related to cell proliferation such as cancer, inflammation, vascular stenosis and angiogenesis by inhibiting CDKs activity. Derivatives, pharmaceutically acceptable salts, hydrates, solvates and isomers thereof and methods of making the same:

[Formula 1]

[Formula 2]

[Formula 3]

In the above formula,

R 1 is hydrogen, alkyl, amino alkyl or hydroxy alkyl group,

R 2 is hydrogen, methyl, trifluoromethyl or halogen,

R3 represents an alkyl, amino alkyl or hydroxy alkyl group,

X represents a halogen.

The term "alkyl" in the above description of substituents of compounds of formula 1, 2 or 3 refers to straight or branched chain saturated hydrocarbon radicals having 1 to 6 carbon atoms such as methyl, ethyl, isopropyl, isobutyl, t-butyl and the like. "Halogen" also means fluorine, chlorine, bromine, iodine.

The compounds of the formula (1), (2) or (3) according to the invention may have asymmetric carbon centers in their structure, so that they may exist as individual enantiomers or diastereomers and also as mixtures thereof including racemates. have. Accordingly, such isomers or mixtures thereof are also within the scope of the present invention.

Representative compounds of the invention include compounds as indicated below.

1. 2- (3-Chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one.

2. 2- (3-Chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (4-pyridinyl) -4 H -chromen-4-one.

3. 4- [2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1-methylpyridinium bromide.

4. 2- (4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one.

5. 3-hydroxy-2- (4-hydroxy-3-methylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one.

6. 3-hydroxy-2- (4-hydroxy-3-trifluoromethylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one .

7. 2- (3-Chloro-4-hydroxyphenyl) -3-hydroxy-8- [1- (2-hydroxyethyl) -4-piperidinyl] -6-methyl-4 H -chromen -4-one.

8. 8- [1- (2-Aminoethyl) -4-piperidinyl] -2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl- 4H -chromen- 4-on.

The compounds of formula 1, 2 or 3 according to the invention may also form pharmaceutically acceptable salts. 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 acid such as citric acid, acetic acid, trichloroacetic acid or trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., sulfonic acid such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid Acid addition salts formed by and the like are included.

Compounds of formula (1), (2) or (3) according to the present invention are reacted with phenylethanone of formula (4) with benzaldehyde of formula (5) to obtain propene of formula (6), and the compound is cyclized to obtain chromenone of formula (7). Protecting the hydroxy group in the compound to obtain a chromanone of formula (8), reacting this compound with 4-bromopyridine to obtain a chromamenon of formula (9), and then deprotection to obtain a compound of formula (2), or Menon may be reacted with a halogenoalkan to give a compound of formula 10 and deprotected to give a compound of formula 3, or to reduce and deprotect a compound of formula 10 to obtain a compound of formula 1:

[Formula 4]

[Formula 5]]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

[Formula 11]

In the above formula,

R1, R2, R3 and X are as defined above,

R ', R ", and R'" are each a hydroxy protecting group.

If the synthesis method is specifically given by way of example, the following schemes 1, 2 and 3.

Scheme 1

Scheme 2

Scheme 3

In the above formula,

R1, R2, R3, X, R ', R ", R'" are as defined above.

In Scheme 1, the compound of Formula 4 may be reacted with benzaldehyde of Formula 5 and 3 equivalents of sodium hydroxide in an 80% ethanol aqueous solution at room temperature for 24 hours to obtain a compound of Formula 6. This compound may be reacted with 3 equivalents of 10% aqueous sodium hydroxide solution and 3 equivalents of hydrogen peroxide at room temperature for 20 hours in a methanol solvent to obtain a compound of Formula 7. This compound was reacted for 30 minutes at room temperature with 2 equivalents of iodomethane, potassium carbonate and N, N-dimethylformamide as a solvent to obtain the compound of formula 8. Compound (8), 1.5 equivalents of bispinacolatodiborone, 3 equivalents of potassium acetate, and 5 mole percent dichlorobistriphenylphosphine palladium were heated to 80 ° C. to 90 ° C. in a nitrogen environment with N, N-dimethylformamide solvent. And react for 2 hours. After cooling to room temperature, 2 equivalents of 4-bromopyridine hydrochloride, 5 mol% dichlorobistriphenylphosphine palladium, and 5 equivalents of 2 mol sodium carbonate aqueous solution were added and reacted for 15 hours to obtain a compound of Formula 9. This compound is heated to reflux with acetone or acetonitrile for 5 hours with 2 equivalents of iodomethane to obtain a compound of formula 10. The compound of Formula 10 is dissolved in 50% methanol / dichloromethane, 5 mol% platinum oxide is added, and the compound of Formula 11 is obtained by reacting at room temperature and 1 atm hydrogen pressure for 24 hours. The compound was added to dried dichloromethane and 3 equivalents of borontribromide was added and reacted at room temperature for 1 hour to obtain a compound of Formula 1. In Scheme 2, the compound of Formula 9 is added to dried dichloromethane, and 3 equivalents of boron tribromide is added to react at room temperature for 1 hour to obtain a compound of Formula 2. In the reaction scheme 3, the compound of formula 10 is added to dried dichloromethane, and 3 equivalents of borontribromide is added for 1 hour at room temperature to obtain the compound of formula 3.

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

However, the preparation method of the compound according to the present invention is not limited to the above description, and can be easily prepared by arbitrarily combining various synthesis methods described in this specification or published in the prior literature, and such combinations It is a common technique that is generalized to those skilled in the art to which the present invention belongs.

Compounds of the formula (1), (2) or (3) according to the present invention can be usefully used as an agent for inhibiting and treating diseases related to cell proliferation such as cancer, inflammation, vascular stenosis and angiogenesis due to its excellent inhibitory activity against CDKs. Accordingly, the present invention provides a cancer, inflammation, characterized in that it contains a compound of formula (1), (2) or (3), a pharmaceutically acceptable salt, hydrate, solvate or isomer thereof as an active ingredient together with a pharmaceutically acceptable carrier, Another object is to provide a composition for inhibiting and treating diseases related to cell proliferation such as vascular stenosis and angiogenesis.

The total daily dose to be administered to the host in a single dose or in separate doses when administering a compound of the present invention for clinical purposes is preferably in the range of 1 to 50 mg per kilogram of body weight, but the specific dose level for a particular patient is determined by the specific compound to be used, Weight, sex, health status, diet, time of administration, method of administration, rate of excretion, drug mixing and the severity of the disease may vary.

The compounds of the present invention can be administered in injectable and oral formulations as desired.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing agents, wetting agents or suspending agents according to known techniques. Solvents that can be used include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are conventionally employed as a solvent or suspending medium. Any non-irritating fixed oil may be used for this purpose, including mono- and diglycerides, and fatty acids such as oleic acid may be used in the preparation of injectables.

Solid dosage forms for oral administration may be capsules, tablets, pills, powders and granules, and capsules and tablets are particularly useful. Tablets and pills are preferably prepared with enteric agents. The solid dosage form is prepared by mixing the active compound of formula 1, 2 or 3 according to the invention with a carrier selected from one or more inert diluents such as sucrose, lactose, starch and the like, lubricants such as magnesium stearate, disintegrants and binders Manufacture.

When the compound of the present invention is to be clinically administered to obtain a desired anticancer effect, the active compound of Formula 1, 2 or 3 may be administered simultaneously with at least one component selected from known anticancer agents. Anticancer agents that can be administered in combination with the compounds of the present invention in this manner include 5-fluorouracil, cisplatin, doxorubicin, taxol, gemcitabine, and the like.

However, the compound-containing preparation according to the present invention for the purpose of anticancer effect is not limited to those described above, and any agent useful for the treatment and prevention of cancer may be included.

The present invention is explained in more detail by the following examples and experimental examples, but the scope of the present invention is not limited in any way by these.

Preparation Example 1 ( E ) -1- (3-Bromo-2-hydroxy-5-methylphenyl) -3- (3-chloro-4-methoxyphenyl) -2-propen-1-one synthesis

150 g of 80% ethanol aqueous solution of 10 g (43.7 mmol) of 1- (3-bromo-2-hydroxy-5-methylphenyl) -1-ethanone and 9.0 g (1.2 molar equivalent) of 3-chloro-4-methoxybenzaldehyde To this was added 5.2 g (3.0 molar equivalents) of sodium hydroxide (NaOH) and stirred at room temperature for 17 hours. After the reaction was completed, neutralized with 2N aqueous hydrochloric acid solution and diluted with 150ml of water. At this time, a yellow solid was precipitated and washed with water and methanol to obtain 14.2 g (yield 85%) of the title compound.

¹ H NMR (CDCl ₃ , ppm): 7.88 (1H, d), 7.78 (1H, s), 7.68 (1H, s), 7.63 (1H, s), 7.54 (2H, m), 6.87 (1H, d ), 3.98 (3H, s), 2.37 (3H, s).

Mass (m / e) = 381 [M ⁺ +1] ⁺

Preparation Example 2 Synthesis of 8-bromo-2- (3-chloro-4-methoxyphenyl) -3-hydroxy-6-methyl- 4H -chromen-4-one.

14.2 g (37.2 mmol) of the compound synthesized in Preparation Example 1 was added to 200 ml of methanol, and 55.8 ml (3 equivalents) of 2 mol aqueous sodium hydroxide solution and 12.6 ml (3 equivalents) of 34% hydrogen peroxide solution were added thereto. After stirring for 3 hours at room temperature, the mixture was neutralized with 2 mol hydrochloric acid, diluted with 200 ml of water, and filtered. The obtained solid was washed with water and methanol to obtain 9.17 g (yield 65.7%) of the desired compound.

¹ H NMR (CDCl ₃ , ppm): 8.43 (1H, s), 8.30 (1H, d), 7.96 (1H, s), 7.77 (1H, s), 7.10 (1H, d), 6.96 (1H, br s), 3.99 (3H, s), 2.46 (3H, s).

Mass (m / e) = 395 [M ⁺ +1] ⁺

Preparation Example 3 Synthesis of 8-Bromo-2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl- 4H -chromen-4-one

9.17 g (24.4 mmol) of the compound synthesized in Preparation Example 2 were heated to reflux for 5 hours in 100 ml of acetone together with 5.2 g (1.5 equivalents) of iodomethane and 4.4 g (1.3 molar equivalents) of potassium carbonate. After completion of the reaction, the mixture was cooled to room temperature and filtered. The obtained solid was washed with water and acetone to obtain 8.0 g (yield 79.8%) of purified target compound.

¹ H NMR (CDCl ₃ , ppm): 8.33 (1H, s), 8.22 (1H, d), 7.97 (1H, s), 7.73 (1H, s), 7.08 (1H, d), 3.99 (3H, s ), 3.92 (3H, s), 2.45 (3H, s).

Mass (m / e) = 409 [M ⁺ +1] ⁺

[Preparation Example 4] 2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl-8- (4-pyridinyl) -4 H - Synthesis of chromene-4-one.

3.0 g (7.32 mmol) of the compound synthesized in Preparation Example 3, 2.8 g of 1.5 equivalents of bispinacolato diborone, 5 mol% dichlorobistriphenylphosphine palladium and 2.15 g of potassium acetate were added to a 20 ml solvent of dimethylformamide. It melt | dissolved and made it react at 80 degreeC under nitrogen gas for 2 hours. After cooling to room temperature, 2 equivalents of 4-bromopyridine hydrochloride, 5 mol% dichlorobistriphenylphosphinepalladium, and 5 equivalents (18.3 ml) of 2M sodium carbonite solution were added, and the mixture was stirred at 80 ° C. for 15 hours in nitrogen gas. The solution was filtered and washed with dimethylformamide and 10% methanol / methylene chloride. The filtrate was concentrated and treated with 30 ml of water, and the resulting pale yellow solid was filtered off. The filtered solid was washed with water and acetone and dried to obtain 2.1 g (yield 70%) of the title compound.

¹ H NMR (CDCl ₃ , ppm): 8.78 (2H, d), 8.12 (1H, s), 8.06 (1H, s), 7.91 (1H, d), 7.55 (3H, m), 6.97 (1H, d ), 3.96 (3H, s), 3.94 (3H, s), 2.52 (3H, s).

Mass (m / e) = 408 [M ⁺ +1] ⁺

Preparation Example 5 4- [2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1-methylpyri Synthesis of Dinium Iodine.

2.1 g (51.5 mmol) of the compound obtained in Preparation Example 4 were refluxed with 2 equivalents of iodomethane in a 30 ml acetonitrile solvent for 3 hours. The solution was concentrated, treated with methanol, filtered and washed with methanol solvent to give 2.44 g (yield 86%) of the title compound.

¹ H NMR (DMSO-d ₆ , ppm): 9.15 (2H, d), 8.57 (2H, d), 8.13 (1H, s), 7.97 (2H, m), 7.88 (1H, d), 7.31 (1H) d), 4.41 (3H, s), 3.95 (3H, s), 3.85 (3H, s), 2.54 (3H, s).

Mass (m / e) = 423 [M ⁺ ]

[Preparation Example 6] 2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen-4 Synthesis of On.

2.44 g (4.43 mmol) of the compound obtained in Preparation Example 5 were reacted for 48 hours in 1 mol of hydrogen in 5 mol% Pt 2 O, 50% methanol / dichloromethane solvent. After filtration through a pad of celite, the mixture was concentrated to give 2.3 g (yield 93%) of the title compound in the form of iodide.

¹ H NMR (CDCl ₃ , ppm): 8.09 (1H, d), 7.95 (2H, s), 7.44 (1H, s), 7.10 (1H, d), 4.00 (3H, s), 3.93 (3H, s ), 3.77 (2H, m), 3.57 (1H, m), 3.03 (2H, m), 2.85 (3H, s), 2.74 (2H, m), 2.46 (3H, s), 2.23 (2H, m) .

Mass (m / e) = 428 [M ⁺ +1] ⁺

Example 1 2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen-4 Synthesis of On.

2.3 g (4.13 mmol) of the compound obtained in Preparation Example 6 were dissolved in 50 ml of dichloromethane, 5 equivalents of boron tribromide was added and reacted at room temperature for 10 hours. The remaining boron tribromide was decomposed with methanol, and then concentrated under reduced pressure. The resulting solid was washed with 10% methanol / dichloromethane to obtain 1.93 g (yield 97%) of the yellow target compound in the form of bromate.

¹ H NMR (DMSO-d ₆ , ppm): 9.60 (1H, br s), 8.23 (1H, s), 8.01 (1H, d), 7.77 (1H, s), 7.47 (1H, s), 7.19 ( 1H, d), 5.75 (1H, s), 3.61-3.42 (5H, m), 2.87 (3H, s), 2.44 (3H, s), 2.05 (2H, m), 1.90 (2H, m).

Mass (m / e) = 400 [M ⁺ +1] ⁺

Example 2 2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (4-pyridinyl) -4 H - Synthesis of chromene-4-one.

145 mg (0.357 mmol) of the compound synthesized in Preparation Example 4 were tested in the same manner as in Example 1, to obtain 156 mg (yield 94.8%) of the title compound in the form of bromate.

¹ H NMR (CD ₃ OD, ppm): 10.90 (1H, br s), 9.80 (1H, br s), 8.99 (2H, s), 8.20 (2H, s), 8.07 (1H, s), 8.02 ( 1H, s), 7.84 (2H, m), 7.09 (1H, d), 2.49 (3H, s).

Mass (m / e) = 380 [M ⁺ +1] ⁺

Example 3 4- [2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1-methylpyri Synthesis of Dinium Bromide.

30 mg (0.054 mmol) of the compound synthesized in Preparation Example 5 were tested in the same manner as in Example 1, to obtain 15 mg (yield 58%) of the title compound in the form of bromate.

¹ H NMR (DMSO-d ₆ , ppm): 10.94 (1H, s), 9.82 (1H, s), 9.17 (2H, d), 8.56 (2H, d), 8.12 (2H, d), 7.92 (1H) , s), 7.75 (1H, d), 7.10 (1H, d), 4.42 (3H, s), 2.53 (3H, s).

Mass (m / e) = 394 [M ⁺ ]

[Preparation Example 7] 2- (4-methoxyphenyl) -3-methoxy-6-methyl-8- (4-pyridinyl) -4 H - Synthesis of chromene-4-one.

500 mg (2.18 mmol) of 1- (3-bromo-2-hydroxy-5-methylphenyl) -1-ethanone and 0.36 g (1.2 molar equivalent) of 4-methoxybenzaldehyde were prepared in Preparation Example 1 and Preparation Example 2 The reaction was carried out in the same manner as in Example 3 and Preparation Example 4 to obtain 417 mg (51% in total yield) of the target compound.

¹ H NMR (CDCl ₃ , ppm): 8.74 (2H, d), 8.12 (1H, s), 7.92 (2H, d), 7.57 (2H, d), 7.50 (1H, s), 6.94 (2H, d ), 3.88 (3H, s), 3.85 (3H, s), 2.50 (3H, s)

Mass (m / e) = 374 [M ⁺ +1] ⁺

[Preparation Example 8] 2- (4-methoxyphenyl) -3-methoxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - Synthesis of chromene-4-one.

100 mg (0.268 mmol) of the compound obtained in Preparation Example 7 were reacted in the same manner as in Preparation Example 5 and Preparation Example 6, whereby 108 mg (total yield 85%) of the target compound was obtained in the form of iodide.

¹ H NMR (DMSO-d ₆ , ppm): 8.16 (2H, d), 7.86 (1H, s), 7.49 (1H, s), 6.97 (2H, d), 3.90 (3H, s), 3.89 (3H , s), 3.60 (3H, m), 2.91 (5H, m), 2.52 (3H, s), 2.22-2.20 (4H, m).

Mass (m / e) = 394 [M ⁺ +1] ⁺

Example 4 2- (4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - Synthesis of chromene-4-one.

50 mg (0.095 mmol) of the compound obtained in Preparation Example 8 were reacted in the same manner as in Example 1, to obtain 35 mg (yield 82%) of the title compound in the form of bromate.

¹ H NMR (CD ₃ OD, ppm): 8.16 (2H, d), 7.86 (1H, s), 7.49 (1H, s), 6.97 (2H, d), 3.67 (3H, m), 3.35 (2H, m), 2.91 (3H, s), 2.47 (3H, s), 2.26 (2H, m), 2.07 (2H, m).

Mass (m / e) = 366 [M ⁺ +1] ⁺

[Preparation Example 10] 2- (3-methyl-4-methoxyphenyl) -3-methoxy-6-methyl-8- (4-pyridinyl) -4 H - Synthesis of chromene-4-one.

500 mg (2.18 mmol) of 1- (3-bromo-2-hydroxy-5-methylphenyl) -1-ethanone and 0.4 g (1.2 molar equivalent) of 3-methyl-4-methoxybenzaldehyde were prepared in Preparation Example 1 In the same manner as in Example 2, Preparation Example 3, and Preparation Example 4, 252 mg (total yield 30%) of the target compound was obtained.

¹ H NMR (CDCl ₃ , ppm): 8.76 (2H, d), 8.13 (1H, s), 7.90 (1H, d), 7.76 (1H, s), 7.56 (2H, d), 7.52 (1H, s ), 6.87 (1H, d), 3.90 (3H, s), 3.89 (3H, s), 2.52 (3H, s)

Mass (m / e) = 388 [M ⁺ +1] ⁺

[Preparation Example 10] 2- (3-methyl-4-methoxyphenyl) -3-methoxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen-4 Synthesis of On.

252 mg (0.65 mmol) of the compound obtained in Preparation Example 9 were reacted in the same manner as in Preparation Example 5 and Preparation Example 6, to obtain 178 mg (yield 51%) of the target compound in the form of iodide.

¹ H NMR (CDCl ₃ + CD ₃ OD, ppm): 7.90 (1H, s), 7.86 (1H, d), 7.77 (1H, s), 7.40 (1H, s), 6.96 (1H, d), 3.89 (3H, s), 3.80 (3H, s), 3.64 (2H, m), 3.50 (1H, m), 2.91 (2H, m), 2.76 (3H, s), 2.42 (3H, s), 2.26 ( 3H, s), 2.22-2.20 (4H, m).

Mass (m / e) = 408 [M ⁺ +1] ⁺

Example 5 3-hydroxy-2- (4-hydroxy-3-methylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen-4-one Synthesis.

178 g (0.332 mmol) of the compound obtained in Preparation Example 10 were tested in the same manner as in Example 1, to obtain 115 mg (yield 75%) of the title compound in the form of bromate.

¹ H NMR (CD ₃ OD, ppm): 8.02 (1H, s), 7.96 (1H, d), 7.85 (1H, s), 7.50 (1H, s), 6.92 (1H, d), 3.69 (3H, m), 3.44 (2H, m), 2.98 (3H, s), 2.48 (3H, s), 2.30 (5H, m), 2.10 (2H, m).

Mass (m / e) = 380 [M ⁺ +1] ⁺

[Preparation Example 11] 2- (3-trifluoromethyl-4-methoxyphenyl) -3-methoxy-6-methyl-8- (4-pyridinyl) -4 H - chromen-4-one in synthesis.

500 mg (2.18 mmol) of 1- (3-bromo-2-hydroxy-5-methylphenyl) -1-ethanone and 0.4 g (1.2 molar equivalent) of 3-methyl-4-methoxybenzaldehyde were prepared in Preparation Example 1 In the same manner as in Example 2, Preparation Example 3, and Preparation Example 4, 252 mg (total yield 30%) of the target compound was obtained.

¹ H NMR (CDCl ₃ , ppm): 8.75 (2H, d), 8.20 (2H, m), 8.10 (1H, s), 7.50 (4H, m), 7.05 (1H, d), 3.95 (3H, s ), 3.92 (3H, s), 2.51 (3H, s).

Mass (m / e) = 442 [M ⁺ +1] ⁺

[Preparation Example 12] 2- (3-trifluoromethyl-4-methoxyphenyl) -3-methoxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen Synthesis of 4-one.

100 mg (0.226 mmol) of the compound obtained in Preparation Example 11 were reacted in the same manner as in Preparation Example 5 and Preparation Example 6, thereby obtaining 100 mg of the target compound (total yield 75%) in the form of iodide.

¹ H NMR (DMSO-d ₆ , ppm): 8.64 (1H, s), 8.25 (1H, s), 7.81 (1H, s), 7.45 (1H, s), 7.22 (1H, d), 4.00 (3H , s), 3.96 (3H, s), 3.65-3.30 (5H, m), 2.89 (3H, s), 2.46 (3H, s), 2.21 (2H, m), 1.94 (2H, m).

Mass (m / e) = 462 [M ⁺ +1] ⁺

Example 6 3-hydroxy-2- (4-hydroxy-3-trifluoromethylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen- Synthesis of 4-ones.

100 mg (0.169 mmol) of the compound obtained in Preparation Example 12 were tested in the same manner as in Example 1, to obtain 69 mg (yield 79%) of the title compound in the form of bromate.

¹ H NMR (DMSO-d ₆ , ppm): 9.60 (1H, br s), 9.39 (1H, s), 8.66 (1H, s), 8.30 (1H, d), 7.79 (1H, s), 7.46 ( 1H, s), 6.84 (1H, d), 4.39 (1H, m), 3.65-3.33 (4H, m), 2.92 (3H, s), 2.43 (3H, s), 2.32 (2H, m), 2.14 (2H, m).

Mass (m / e) = 434 [M ⁺ +1] ⁺

Preparation Example 13 4- [2- (3-Chloro-4-methoxyphenyl) -3-methoxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1- (2 Synthesis of methoxyethyl) pyridinium bromide.

50 mg (0.122 mmol) of the compound obtained in Preparation Example 4 were refluxed with 2 equivalents of 2-bromoethylmethyl ether in a 5 ml acetonitrile solvent for 3 hours. The solution was concentrated, treated with methanol, filtered and washed with methanol solvent to give 52 mg (yield 78%) of the title compound.

¹ H NMR (DMSO-d ₆ , ppm): 9.15 (2H, d), 8.57 (2H, d), 8.13 (1H, s), 7.97 (2H, m), 7.88 (1H, d), 7.31 (1H) , d), 4.41 (3H, s), 3.95 (6H, d), 3.85 (3H, s), 3.80 (2H, t), 2.54 (3H, s).

Mass (m / e) = 466 [M ⁺ ] ms = 545

Production Example 14 2- (3-Chloro-4-methoxyphenyl) -3-methoxy-8- [1- (2-methoxyethyl) -4-piperidinyl] -6-methyl-4 H -Synthesis of Chromium-4-one.

45 mg (0.082 mmol) of the compound obtained in Preparation Example 13 were reacted in the same manner as in Preparation Example 6, to obtain 30 mg of the target compound (yield 66%) in the form of bromate.

¹ H NMR (CDCl ₃ , ppm): 8.09 (1H, d), 7.95 (2H, s), 7.44 (1H, s), 7.10 (1H, d), 4.00 (3H, s), 3.93 (6H, d ), 3.80 (2H, t), 3.77 (4H, m), 3.57 (1H, m), 3.03 (2H, m), 2.85 (3H, s), 2.74 (2H, m), 2.46 (3H, s) , 2.23 (2H, m).

Mass (m / e) = 472 [M ⁺ +1] ⁺

Example 7 2- (3-Chloro-4-hydroxyphenyl) -3-hydroxy-8- [1- (2-hydroxy-ethyl) -4-piperidinyl] -6-methyl-4 Synthesis of H -chromen-4-one.

25 mg (0.045 mmol) of the compound obtained in Preparation Example 14 were reacted in the same manner as in Example 1, to obtain 15 mg (yield 65%) in the form of bromate.

¹ H NMR (DMSO-d ₆ , ppm): 9.60 (1H, br s), 8.23 (1H, s), 8.01 (1H, d), 7.77 (1H, s), 7.47 (1H, s), 7.19 ( 1H, d), 5.75 (1H, s), 3.61-3.42 (7H, m), 2.87 (2H, m), 2.44 (3H, s), 2.05 (2H, m), 1.90 (2H, m).

Mass (m / e) = 430 [M ⁺ +1] ⁺

Production Example 15 4- [2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1- [2 Synthesis of-(1,3-dioxo-1,3-dihydro- 2H -isoindol-2-yl) ethyl] pyridinium bromide.

50 mg (0.122 mmol) of the compound obtained in Preparation Example 4 were reacted with 2 equivalents of N- (2-bromoethyl) phthalimide in the same manner as in Preparation Example 13 to obtain 60 mg (yield 74%) of the title compound.

¹ H NMR (DMSO-d ₆ , ppm): 9.15 (2H, d), 8.57 (2H, d), 8.13 (1H, s), 7.97 (2H, m), 7.88 (1H, d), 7.55-7.40 (4H, m), 7.31 (1H, d), 4.41 (2H, m), 3.95 (3H, s), 3.85 (3H, s), 2.54 (3H, s), 2.23 (2H, t).

Mass (m / e) = 581 [M +] ms = 660

Production Example 16 2- (2- {4- [2- (3-chloro-4-methoxyphenyl) -3-methoxy-6-methyl-4-oxo- 4H -chromen-8-yl ] -1-piperidinyl} ethyl) -1 H -isoindole-1,3 ( 2H ) -dione.

50 mg (0.075 mmol) of the compound obtained in Preparation Example 15 were reacted in the same manner as in Preparation Example 6, to obtain 45 mg (yield 89%) of the target compound in the form of bromate.

¹ H NMR (DMSO-d ₆ , ppm): 8.09 (1H, d), 7.95 (2H, s), 7.58-7.47 (4H, m), 7.44 (1H, s), 7.10 (1H, d), 4.00 (3H, s), 3.93 (3H, s), 3.77 (2H, m), 3.57 (1H, m), 3.03 (2H, m), 2.85 (2H, s), 2.74 (2H, m), 2.46 ( 3H, s), 2.23 (4H, m).

Mass (m / e) = 587 [M ⁺ +1] ⁺

Production Example 17 2- (2- {4- [2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-4-oxo- 4H -chromen-8-yl ] -1-piperidinyl} ethyl) -1 H -isoindole-1,3 ( 2H ) -dione.

40 mg (0.060 mmol) of the compound obtained in Preparation Example 16 was reacted in the same manner as in Example 1, to obtain 25 mg (yield 65%) of the bromate form.

¹ H NMR (DMSO-d ₆ , ppm): 9.60 (1H, br s), 8.23 (1H, s), 8.01 (1H, d), 7.77 (1H, s), 7.62-7.49 (4H, m), 7.47 (1H, s), 7.19 (1H, d), 5.75 (1H, s), 3.61-3.42 (5H, m), 2.87 (2H, t), 2.44 (3H, s), 2.23 (2H, t) , 2.05 (2H, m), 1.90 (2H, m).

Mass (m / e) = 559 [M ⁺ +1] ⁺

[Example 8] 8- [1- (2-aminoethyl) -4-piperidinyl] -2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl -4 H - Synthesis of Chromen-4-one.

25 mg (0.040 mmol) of the compound obtained in Preparation Example 17 was added to 5 ml of ethanol, and 3 equivalents of hydrazine were added and heated at 60 ° C. for 1 hour. After cooling to room temperature, an aqueous solution of sodium carbonate was added thereto, and the produced solid was filtered to obtain 12 mg (yield 70%) of the title compound.

¹ H NMR (DMSO-d ₆ , ppm): 9.60 (1H, br s), 8.23 (1H, s), 8.01 (1H, d), 7.77 (1H, s), 7.47 (1H, s), 7.19 ( 1H, d), 5.75 (1H, s), 3.61-3.42 (5H, m), 2.87 (2H, t), 2.55 (2H, t), 2.44 (3H, s), 2.05 (2H, m), 1.90 (2H, m).

Mass (m / e) = 429 [M ⁺ +1] ⁺

Experimental Example 1) Inhibitory Activity of CDK2 and CDK4

Analysis of the enzymatic inhibitory effect of CDK2,4 was performed according to the methods of Kitagawa and Carson (Kitagawa, M. et al .; Oncogene, 9: 2549, 1994; Carlson, BA et al .; Cancer Research 56: 2473, 1996). Was performed. The activated CDK2 enzyme was expressed and purified as a combination of histidine-labeled human CDK2 protein and Cyclin A protein, and baculovirus expressing His-CDK2 gene and baculovirus expressing Cyclin A gene. Activators of 14 nmole / min / mg unit activity, 22 mM of Km for ATP, purified from insect cells infected with the virus at the same time, were used. The activated CDK4 enzyme was expressed and purified in insect cells as a combination of human CDK4 protein and CyclinD1 protein linked to GST (Glutathion-S-transferase), and the activity of 57 nmole / min / mg and 940 uM of KM for ATP was detected. Enzymes were used. Substrates of both enzymes were purified using the C-terminal region protein having the substrate specificity of human Rb protein, and the concentration of [gamma-32P] in the presence of a compound diluted by concentration, an appropriate amount of CDK2 / cyclinA or CDK4 / cyclinD1 and the substrate protein. labeled] ATP was added to the reaction and the substrate was separated to measure the radioactivity contained in the substrate.

According to the method described above, the inhibitory capacity of each enzyme activity of the inhibitor according to the present invention, which was measured for CDK2 and CDK4, was expressed as an IC50 value. The results are as shown in Table 1 below.

TABLE 1

Inhibitory Activity of CDK2 and CDK4

Table 1 continued)

Experimental Example 2) Acute Toxicity Test

In order to investigate the acute oral toxicity of the compounds obtained in Examples 1 and 3, a solution containing the compound in various concentrations was orally administered to male mice of the ICR line at a dose of 10 ml per kg. The mortality and symptoms for 7 days after oral administration were observed and the median lethal dose (LD50, mg / kg) was calculated according to the Litchfield-Wilcoxon method and the results are shown in Table 2.

TABLE 2

Test compound LD50 (mg / kg) Example 1 3,000 Example 3 3,000

Claims (4)

A compound of Formula 1, 2 or 3, a pharmaceutically acceptable salt, hydrate, solvate or isomer thereof: [Formula 1] [Formula 2] [Formula 3] In the above formula, R1 is hydrogen, straight or branched chain saturated hydrocarbon of 1 to 6 carbon atoms, amino alkyl or alkyl group of straight or branched chain saturated hydrocarbon of 1 to 6 carbon atoms, or hydroxy alkyl group of straight or branched chain saturated hydrocarbon of 1 to 6 carbon atoms Is, R 2 is hydrogen, methyl, trifluoromethyl or halogen, R 3 represents a straight or branched chain saturated hydrocarbon having 1 to 6 carbon atoms, an amino alkyl or alkyl group having 1 to 6 carbon atoms straight or branched chain saturated hydrocarbon, or a hydroxy alkyl group having 1 to 6 straight chain or branched chain saturated hydrocarbons. , X represents a halogen. The method of claim 1, 1. 2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one, 2. 2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (4-pyridinyl) -4 H -chromen-4-one, 3. 4- [2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl-4-oxo- 4H -chromen-8-yl] -1-methylpyridinium bromide, 4. 2- (4-hydroxyphenyl) -3-hydroxy-6-methyl-8- (1-methyl-4-piperidinyl) -4 H - chromen-4-one, 5. 3-hydroxy-2- (4-hydroxy-3-methylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one, 6. 3-hydroxy-2- (4-hydroxy-3-trifluoromethylphenyl) -6-methyl-8- (1-methyl-4-piperidinyl) -4 H -chromen-4-one , 7. 2- (3-Chloro-4-hydroxyphenyl) -3-hydroxy-8- [1- (2-hydroxyethyl) -4-piperidinyl] -6-methyl-4 H -chromen -4-one, and 8. 8- [1- (2-Aminoethyl) -4-piperidinyl] -2- (3-chloro-4-hydroxyphenyl) -3-hydroxy-6-methyl- 4H -chromen- Compound selected from the group consisting of 4-ons. An anticancer composition comprising a compound of the formula (1), (2) or (3) as defined in claim 1, a pharmaceutically acceptable salt, hydrate, solvate or isomer thereof as an active ingredient with a pharmaceutically acceptable carrier. Phenylethanone of Formula 4 is reacted with benzaldehyde of Formula 5 to obtain propenone of Formula 6, and the compound is cyclized to obtain a chromen of Formula 7, and the hydroxy group is protected from the compound to obtain a chromenone of Formula 8 , By reacting the compound with 4-bromopyridine to obtain the chromonone of formula 9, and then deprotecting to obtain the compound of formula 2, or by reacting the chromenone of formula 9 with halogenoalkanes to give the compound of formula 10 Obtaining, deprotecting to obtain a compound of formula 3, or reducing the compound of formula 10 and deprotecting to obtain a compound of formula 1, wherein the compound of formula 1, 2 or 3 according to claim 1 How to manufacture: [Formula 4] [Formula 5]] [Formula 6] [Formula 7] [Formula 8] [Formula 9] [Formula 10] [Formula 11] In the above formula, R1, R2, R3 and X are as defined in claim 1, R ', R ", R'" are each a hydroxy protecting group.