KR101713638B1 - Novel pyrrolo pyrimidine derivatives and composition for preventing or treating cancer comprising the same - Google Patents

Abstract

상기 식에서, R¹, R², R³, R⁴, R⁵ 및 X는 명세서 중에 정의한 바와 같다.The present invention relates to a novel pyrrolopyrimidine compound represented by the following formula 1, a pharmaceutically acceptable salt or hydrate thereof, a process for producing the same, and a pharmaceutical composition for preventing or treating cancer comprising the compound.
[Chemical Formula 1]

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and X are as defined in the specification.

Description

[0001] The present invention relates to novel pyrrolopyrimidine derivatives and their use for preventing or treating cancer,

The present invention relates to a novel pyrrolopyrimidine compound represented by the following formula 1, a pharmaceutically acceptable salt or hydrate thereof, a process for producing the same, and a pharmaceutical composition for preventing or treating cancer comprising the compound.

[Chemical Formula 1]

Heat shock proteins (Hsps) are molecular chaperons whose expression is increased by various protein toxic stressors such as heat, hypoxia, heavy metals and acid hypertrophy. They protect cells from damage caused by stress factors, Of the world economy. Heat shock proteins (Hsps) are molecular chaperones that support the folding and misfolding of normal proteins under normal conditions or the prevention of aggregation of unfolded proteins. They are responsible for the regulation of several key functions of cells, cell proliferation and cell apoptosis It is important.

Families of heat shock proteins (Hsps) are classified according to molecular weight, such as Hsp100, Hsp90, Hsp70, Hsp60 and Hsp27. Among them, the heat shock protein 90 kDa (Hsp90) is an ATP-dependent molecular chaperone that occupies 1-2% of the total cell protein mass under normal conditions, and Hsp90α and Hsp90β (cytoplasm), Grp94 (endoplasmic reticulum) and TRAP-1 (Mitochondria). Hsp90 plays an important role in promoting structural maturation, stabilization and activation of more than 300 proteins called client proteins, or playing an important role in managing client proteins through proteolysis and adapting to intracellular signaling and stress responses.

The Hsp90 client proteins include steroid hormone receptors (SHRs), protein kinases and the transcription factor p53, including more than 40 tumors such as ErbB2 / Her2, Braf, Akt / PKB, mutant p53, HIF-1α and hTERT Proteins, which are overexpressed in mutant forms in cancer and are responsible for cancer cell proliferation and survival. Hsp90 is overexpressed at levels 2-10 times higher in tumor cells than in normal cells. High activity of Hsp90 is due to overexpression or proliferation of mutant client proteins, and it participates in tumorigenesis processes such as mutation protein stabilization and metastasis . Cancer cells use the chaperone function of Hsp90 to protect against misfolding and degradation of mutated or overexpressed client proteins.

Inhibition of Hsp90 interrupts the interaction between the M-domain of Hsp90 and the client protein, leading to the degradation of the client protein through the ubiquitin proteasome pathway leading to inhibition of cell growth and loss of function. The Hsp90 inhibitor has a binding affinity of about 100 times higher than that of normal cells for Hsp90 derived from tumor cells and can specifically target the inhibition in tumor cells without inhibiting Hsp90 function in normal cells. As such, Hsp90 is attracting attention as a major target for the development of anticancer drugs.

Geldanamycin and radicicol were identified as inhibitors of Hsp90. However, geladinamycin had low solubility and high toxicity, and radicicol had low biological stability. In vivo, There is a problem that it does not show tumor activity. Also, based on the above geladamycin, geladamycin analog 17-AAG (tanespimycin, Kosan-BMS, WO 2005112952) has been synthesized, but there still remains a problem related to hepatotoxicity.

In addition, several Hsp90 inhibitors are in clinical trials, but there is no Hsp90 inhibitor that fully satisfies formulation, low toxicity, high water solubility, and stability, and it is time to develop more advanced Hsp90 inhibitors.

Under these circumstances, the present inventors have continued studies to solve the above problems, and as a result, they have found that, as an anticancer compound exhibiting excellent inhibitory activity against cancer cells and having a markedly low toxicity in normal cells, , A pharmaceutically acceptable salt thereof, or a hydrate thereof, a process for producing the same, and a pharmaceutical composition for the prevention or treatment of cancer comprising the same as an active ingredient.

It is an object of the present invention to provide a novel pyrrolopyrimidine compound, a pharmaceutically acceptable salt or hydrate thereof.

It is another object of the present invention to provide a novel pyrrolopyrimidine compound, a pharmaceutically acceptable salt or hydrate thereof.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating cancer comprising a novel pyrrolopyrimidine compound, a pharmaceutically acceptable salt or hydrate thereof as an active ingredient.

In order to achieve the above object, in a specific example of the present invention, the anticancer effect against cancer cells was verified, and as a result, it was confirmed that there was an effect of inhibiting the proliferation of excellent cancer cells for each cancer cell (Experimental Example 1).

In another embodiment of the present invention, the cytotoxicity of the pyrrolopyrimidine compound prepared in the present invention was measured in normal cells. As a result, it was confirmed that the normal cell did not show cytotoxicity (Experimental Example 2).

In another embodiment of the present invention, the degradation of the client protein and the Hsp70 inducing effect were confirmed, and it was confirmed that there was no change in the expression of Hsp90. Thus, it was confirmed that the compound of the present invention inhibits the function of Hsp90 to exhibit anticancer effect Experimental Example 3).

As described above, the novel pyrrolopyrimidine compound according to the present invention has a high inhibitory activity and antiproliferative effect on cancer cells as a function suppressing agent for Hsp90, and thus it can be used as a composition for preventing or treating cancer .

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention relates to a pyrrolopyrimidine compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof:

[Chemical Formula 1]

In Formula 1,

R ¹ is H; A halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine; Trifluoromethyl; C ₁ -C ₃ alkyl, and C ₁ -C ₅ alkoxy groups,

R ² , R ³ , R ⁴ and R ⁵ are each H; A halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine; A C ₁ -C ₆ alkyl group; A C ₁ -C ₅ alkoxy group and a dialkoxy group connected with a C ₁ -C 3 chain,

X is carbon or nitrogen,

When X is carbon, X is H; Or a halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine.

The C ₁ -C ₃ alkyl group or C ₁ -C ₆ alkyl group may be substituted by C ₁ -C ₃ alkyl; C ₁ -C ₃ alkoxy; C ₆ -C ₁₀ Aryl; A halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine; Phenyl unsubstituted or substituted with C ₁ -C ₃ alkoxy; And an aromatic or nonaromatic group selected from the group consisting of pyridine and pyridine.

In the definition of substituents in the present invention, the term " alkyl " means an aliphatic hydrocarbon radical. Alkyl may be a "saturated alkyl" containing no alkenyl or alkynyl moiety, or an "unsaturated alkyl" containing at least one alkenyl or alkynyl moiety. "Alkenyl" means a group comprising at least one carbon-carbon double bond, and "alkynyl" means a group comprising at least one carbon-carbon triple bond. Alkyl, when used alone or in combination with alkoxy, may be branched or straight chain, respectively.

The term " aryl ", alone or in combination with other radicals, refers to a carbocyclic ring containing 6 carbon atoms which may be further fused with a second 5 or 6 membered carbocyclic group which may be aromatic, Aromatic monocyclic group. Examples of aryl include phenyl, indanyl, 1-naphthyl, 2-naphthyl, tetrahydronaphthyl, and the like. The aryl may be connected to another group at an appropriate position on the aromatic ring, but is not limited thereto.

The term " alkoxy " means an alkyl group (i.e., -O-alkyl) attached to another group through an oxygen atom. The alkoxy group may be unsubstituted or substituted with one or more suitable substituents. Examples of the alkoxy group include a (C1-C6) alkoxy group such as -O-methyl, -O-ethyl, -O-propyl, -O-isopropyl, -0-2-methyl-1-butyl, -0-3-methyl-1-butyl, -0-2-methyl-3-butyl, -0-2,2-dimethyl- Propyl, -0-2-methyl-1-pentyl, 3-O-methyl-1-pentyl, -Methyl-2-pentyl, -O-4-methyl-2-pentyl, -0-2,2-dimethyl- butyl, -O-butyl, -O-isobutyl, -O-butyl, -O-pentyl, -O-isopentyl, -O-neopentyl and -O-hexyl.

Examples of the pyrrolopyrimidine compound of the formula (1) are as follows.

Pyrrolo [2,3-d] pyrimidin-2-amine (Compound (5-benzyl-4-chloro-7- One);

Pyrrolo [2,3-d] pyrimidin-2-amine (Compound (5-benzyl-4-chloro-7- 2);

(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methylbenzyl) -7H- pyrrolo [2,3- d] pyrimidine- 2-amine (Compound 3);

(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methoxybenzyl) -7H-pyrrolo [2,3- d] pyrimidine -2-amine (Compound 4);

(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3 -d] pyrimidin-2-amine (Compound 5);

(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (3- (trifluoromethyl) benzyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amine (Compound 6);

Pyrrolo [2,3-d] pyrimidin-2-ylmethyl) -7- (4-methoxy- 2-amine (Compound 7);

5,7-Dibenzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (Compound 8);

Pyrido [2,3-d] pyrimidin-2-amine (Compound 9) was prepared in accordance with the general method of Example 1 from 7- (benzo [d] [1,3] dioxol- ;

7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) -quinolin- -Amine (Compound 10); And

Yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) -Amine < / RTI > (Compound 11).

The pyrrolopyrimidine compound according to the present invention may be used in the form of a pharmaceutically acceptable salt or hydrate derived from an inorganic or organic acid. Examples of the salt include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, But are not limited to, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, , Cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, and toluenesulfonic acid. In addition, the hydrate may mean that the compound of formula (1) is formed by bonding with water molecules.

In another aspect, the present invention relates to a pyrrolopyrimidine compound represented by Formula 1, a pharmaceutically acceptable salt or hydrate thereof.

Hereinafter, a specific example of the method for producing the pyrrolopyrimidine compound represented by Formula 1 is described.

Production method: Pirolo  Preparation of pyrimidine compound

As illustrated below, the pyrrolopyrimidine compound of formula (I), a pharmaceutically acceptable salt or hydrate thereof, may be prepared by reacting a compound of formula (II) with a compound of formula (III).

(2)

In Formula 2, R ¹ is H; A halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine; Trifluoromethyl; C ₁ -C ₃ alkyl, and C ₁ -C ₅ alkoxy groups.

 (3)

In Formula 3,

R ² , R ³ , R ⁴ and R ⁵ are each H; A halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine; A C ₁ -C ₆ alkyl group; A C ₁ -C ₅ alkoxy group and a dialkoxy group connected with a C ₁ -C 3 chain,

X is carbon or nitrogen,

Z is chlorine, bromine or iodine,

When X is carbon, X is H; Or a halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine.

The step of reacting the compound of Formula 2 with the compound of Formula 3 can be carried out by the following Reaction Scheme 1.

[Reaction Scheme 1]

The process of Reaction Scheme 1 can be carried out by an alkylation reaction of NH in Formula 2 and Z (Cl, Br or I) in Formula 3 (International Patent Application Publication No. 2005/0113340).

The compound of formula 2 may be prepared by cyclisation of the amide carbonyl formed with a cyclization reaction with [alpha] -bromoaldehyde, as in the following references (U.S. Patent Application Publication No. US2011-0028487, International Patent Application Publication WO2005-028434) .

The compound of Formula 3 is produced in the same manner as in the following references (International Patent Application Publication WO2013-071169, International Patent Application Publication US 2005-0113340, Bogucki, DE et al . J. Org. Chem. 1995, 60, 588) can do.

In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising the pyrrolopyrimidine compound represented by the formula (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof as an active ingredient.

The pharmaceutical composition may be applied to an experimental animal such as a mouse, a rabbit, a rat, a guinea pig, or a hamster, or a primate including a human, but is not limited thereto, and is preferably applied to a primate including a human, more preferably a human.

As used herein, " treatment " is intended to include alleviation or amelioration of symptoms, reduction in the extent of disease, delay or alleviation of disease progression, improvement, alleviation or stabilization of disease states, partial or complete recovery, May be used to mean both. For example, the prevention or treatment of cancer in the present invention includes the treatment and / or amelioration of cancer metastatic and proliferative diseases.

In the present specification, treatment of cancer means treatment for all cancer cells, and cancer includes angiogenesis of endothelial cells and mitosis thereof (solid tumors, tumor metastases and benign tumors). For example, the term cancer refers to breast cancer, ovarian cancer, cervical cancer, prostate cancer, colon cancer, genitourinary cancer, esophageal cancer, laryngeal cancer, glioblastoma, gastric cancer, skin cancer, keratocellular carcinoma, lung cancer, squamous cell carcinoma, Cancer, kidney cancer, myelodysplastic syndrome, lymphoid cancer, colon cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, hepatocellular carcinoma, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, Cancer, colon cancer, colon cancer, colorectal cancer, brain cancer, central nervous system cancer, leukemia, hemangioma, trachoma or pyogenic granulomas. But are not limited thereto.

The pharmaceutically acceptable salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, A salt number of an inorganic or organic acid selected from the group consisting of ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, and toluenesulfonic acid have. In addition, the hydrate may mean that the compound of formula (1) is formed by bonding with water molecules.

Depending on the use of the pharmaceutical composition of the present invention and the method of use, the content of the compound represented by the general formula (1), its pharmaceutically acceptable salt or hydrate thereof as an active ingredient may be appropriately adjusted according to the selection of a person skilled in the art.

For example, the pharmaceutical composition may contain 0.1 to 10% by weight, more preferably 0.5 to 5% by weight, of the compound represented by Formula 1, its pharmaceutically acceptable salt, or hydrate, based on the total weight of the total composition As shown in FIG.

The pharmaceutically acceptable salts or hydrates of the compound represented by the above-mentioned formula (I) may be contained alone in the pharmaceutical composition, or may be contained together with other pharmacologically acceptable carriers, excipients, diluents or subcomponents.

Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , Dextrin, calcium carbonate, propylene glycol, liquid paraffin, and physiological saline. However, it is not limited to these, and any conventional carrier, excipient or diluent may be used. In addition, the pharmaceutical composition may further comprise at least one selected from the group consisting of conventional fillers, extenders, binders, disintegrants, anticoagulants, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, , Fragrances, emulsifiers, preservatives, and the like.

In addition, the pharmaceutical composition may further include one or more other anticancer agents or other therapeutic agents known to be effective for the treatment or prevention of cancer, in addition to the above-mentioned effective ingredients, and may be used as a combination therapy simultaneously or simultaneously. Other anticancer agents or other therapeutic agents which may be applied to the combination therapy include, for example, Gleevec (R), imatinib, Sutent (R), sunitinib, Herceptin (R), Trastuzumab, Velcade (R), Bortezomib, Dexamethasone, an Hsp90 inhibitor, an aromatase inhibitor, or a kinase inhibitor.

The pharmaceutical composition may be administered orally or parenterally. For example, the pharmaceutical composition may be administered through various routes including oral, transdermal, subcutaneous, intravenous, or muscular. In addition, the formulations of the compositions may vary depending on the method of use and may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal . In general, solid dosage forms for oral administration include tablets, pills, soft or hard capsules, pills, powders and granules, which may contain one or more Excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, Sweeteners, fragrances, preservatives, and the like. Forms for parenteral administration include creams, looses, ONITMENTS, PLASTERS, LIQUIDS AND SOULTIONS, AEROSOLS, FRUIDEXTRACTS, Or may be in the form of an isotonic solution (ELIXIR), INFUSIONS, SACHET, PATCH or INJECTIONS, and in case of injectable form, it may preferably be in the form of an isotonic aqueous solution or suspension .

The pharmaceutical composition may further contain a sterilizing agent, an antiseptic, a stabilizer, a wetting agent or an emulsifying accelerator, an auxiliary agent such as a salt and / or a buffer for controlling osmotic pressure, and other therapeutically useful substances, Or coating method, and may be formulated using other appropriate methods known in the art.

The dosage of the pharmaceutical composition may be determined in consideration of the administration method, the age and sex of the recipient, the severity of the patient, the condition, the degree of absorption of the active ingredient in the body, the inactivation rate, Can be administered separately. The active ingredient of the pharmaceutical composition is preferably administered orally or parenterally once daily or divided into mammals, including humans, in an amount of 0.001 to 100 mg / kg body weight, preferably 0.01 to 35 mg / kg body weight per day May be administered via an old route.

Another embodiment of the present invention provides a method for treating cancer, comprising administering a therapeutically effective amount of a compound represented by the following formula 1, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

Preferably, the method of treatment may further comprise identifying a patient in need of prevention or treatment of the cancer prior to the administration step.

"Therapeutically effective amount" means the amount of active ingredient for mammals effective for the prevention or treatment of cancer, and the therapeutically effective amount will vary depending on the type of disease, the severity of the disease, the active ingredient contained in the composition, May be adjusted depending on various factors including the content, the type of the formulation and the age, body weight, general health condition, sex and diet of the patient, administration time, administration route and blood clearance rate of the composition, May be administered orally or parenterally, once or divided once a day, in an amount of 0.001 to 100 mg / kg body weight, preferably 0.01 to 35 mg / kg body weight on a daily basis, as described above .

The pharmaceutical composition for prevention or treatment of the pyrrolopyrimidine compound of the present invention, its pharmaceutically acceptable salt or hydrate and cancer thereof as an active ingredient shows excellent cytotoxicity and exhibits excellent Hsp90 inhibitory activity and anti-cancer effect , Inhibition of cancer as well as cancer chemotherapy can be used.

FIG. 1 shows that when Compound 1 of the present invention was treated with MCF-7 cell lines at different concentrations (0, 1, 5, 10, 20 and 30 uM), Hsp90, a client protein Epidermal growth factor receptor (EGFR) factor receptor 2), Met, and Akt (protein kinase B). In FIG. 1, "0uM" represents 1% DMSO as a control, "GA" represents geladamycin, "HSP70" represents 70 kDa (70 kilodalton heat shock protein) and "B-Actin" represents Beta-actin.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  One. 5-Benzyl-4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -7H-pyrrolo [ 2,3-d] pyrimidine -2-amine (Compound 1)

[Compound 1]

[Total reaction formula]

Step 1. 2-Amino-5-benzyl-3,7- Dihydro -4H- Pyrrolo [2,3-d] pyrimidine -4-one

(5.00 g, 37.26 mmol) was dissolved in a solution of dichloromethane (149 mL) and 1,4-dioxane (15 mL) under an argon atmosphere, and then bromotrimethylsilane (7.38 mL, 55.90 mmol) Lt; / RTI > After stirring at room temperature for 1 hour, bromine (1.91 mL, 37.26 mmol) was added to the reaction solution and stirred for 1 hour. The completion of the reaction was confirmed by TLC (nucleic acid: ethyl acetate = 4: 1). The reaction mixture was poured into water, and the organic solvent layer extracted with dichloromethane (150 mL x 3) was washed again with a saturated sodium chloride solution (500 mL). The organic solvent layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the desired compound g, qaunt. Yield). Bromo-3-phenylpropanal (7.94 g, 37.26 mmol) synthesized in the above experiment and dissolved in acetonitrile (74 mL) under argon atmosphere was treated with 4-diamino-6-hydroxypyrimidine , 37.26 mmol), sodium acetate (6.11 g, 74.53 mmol) and distilled water (74 mL). The reaction mixture was stirred at room temperature for 13 hours, and the completion of the reaction was confirmed by TLC (dichloromethane: methanol = 10: 1). The resulting precipitate was filtered with acetonitrile to obtain the desired compound (2.46 g, 27% yield).

mp 282.4-283.7 C;

¹ H NMR (300 MHz, DMSO- d ₆ ) δ 10.70 (s, 1H), 10.12 (s, 1H), 7.30-7.20 (m, 4H), 7.14-7.09 1H), 6.00 (s, 2H), 3.92 (s, 2H);

¹³ C NMR (75 MHz, DMSO- d ₆ )? 159.8, 152.6, 151.6, 142.8, 129.0, 128.4, 125.9, 118.1, 114.3, 99.2, 32.2.

Step 2. 5-Benzyl-4- Chloro -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

2-amino-5-benzyl-3,7-dihydro-4H-pyrrolo [2,3-d] pyrimidin-4-one (2.46 g, 10.23 mmol) prepared in step (1) Was dissolved in acetic anhydride (11 mL) and refluxed for 3 hours. The reaction solution was concentrated under reduced pressure to remove the solvent. After concentrating the concentrate in acetonitrile (51 mL), benzyltriethylammonium chloride (4.66 g, 20.46 mmol), N, N-dimethylaniline (2.59 mL, 20.46 mmol) and phosphorus (V) oxychloride (11.97 mL, 61.38 mmol) was added thereto, followed by stirring at 100 ° C for 1 hour. The completion of the reaction was confirmed by TLC (dichloromethane: methanol = 10: 1). After the reaction solution was concentrated under reduced pressure to remove the solvent, the concentrate was poured into ice water and neutralized to pH 7 with 2N sodium hydroxide solution. The mixed aqueous solution was extracted with dichloromethane (20 mL × 3). The extracted organic solvent layer was washed with saturated sodium chloride solution (60 mL), and the organic solvent layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was dissolved in methanolic 3N HCl (34 mL) under argon atmosphere and stirred at 50 ° C for 2 hours. The completion of the reaction was confirmed by TLC (dichloromethane: methanol = 10: 1). The reaction solution was cooled to room temperature, neutralized to pH 7 with 2N sodium hydroxide solution, and the resulting precipitate was filtered to obtain the desired compound (2.44 g, 92% yield).

mp 255.0-256.6 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.25 (s, 1H), 7.29-7.16 (m, 6H), 6.79 (s, 1H), 6.44 (s, 2H), 4.04 (s, 2H) ;

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.6, 155.7, 151.3, 141.5, 128.8, 128.6, 126.2, 121.9, 113.2, 107.8, 31.8.

Step 3. 5-Benzyl-4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -7H- blood Lolo [ 2,3-d] pyrimidine -2- Amine  Produce

Benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (300 mg, 1.16 mmol) prepared in the above step (2) was dissolved in N, N- 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (268 mg, 1.21 mmol) and potassium carbonate (481 mg, 3.48 mmol) were added to the solution and the mixture was stirred at 45 ° C for 13 hours And the completion of the reaction was confirmed by TLC (dichloromethane: methanol = 10: 1). The reaction solution was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The concentrate was separated and purified by column chromatography (dichloromethane: methanol = 40: 1) to obtain the desired compound (146 mg, 31% yield).

mp 152.0-153.1 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.22 (s, 1H), 7.30-7.20 (m, 5H), 6.57 (s, 1H), 5.31 (s, 2H), 5.07 (s, 2H), 4.15 (s, 2H), 3.76 (s, 3H), 2.27 (s, 3H), 2.22 (s, 3H);

¹³ C NMR (75 MHz, CDCl ₃ ) δ 164.3, 158.3, 154.1, 154.0, 152.4, 149.5, 140.5, 128.6, 128.3, 126.0, 125.2, 124.5, 114.6, 109.3, 59.9, 47.4, 32.0, 13.3, 10.9.

Example  2. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (3- Methylbenzyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 2)

[Compound 2]

[Total reaction formula]

Step 1. Preparation of 3- (m-tolyl) Propanal  Produce

3-iodotoluene (1.18 mL, 9.17 mmol) was dissolved in N, N-dimethylformamide (16 mL) under an argon atmosphere and benzyltriethylammonium chloride (2.09 g, 9.17 mmol) and sodium bicarbonate (1.54 g, 18.35 mmol), allyl alcohol (1.25 mL, 18.35 mmol) and palladium (II) acetate (494 mg, 2.20 mmol) were added to the reaction mixture and stirred at 40 ° C for 12 hours. TLC (hexane: ethyl acetate = 2: 1). After cooling the reaction solution to room temperature, the reaction mixture was poured into ice water and extracted with dichloromethane (20 mL × 3). The extracted organic solvent layer was washed with saturated sodium chloride solution (60 mL), the organic solvent layer was dried over anhydrous sodium sulfate, To remove the solvent. The concentrate was separated and purified by column chromatography (hexane: ethyl acetate = 7: 1) to obtain the desired compound (912 mg, 67% yield).

^{1 H NMR (300 MHz, CDCl} 3) δ9.84 (s, 1H), 7.28 (t, J = 7.8 Hz, 2H), 7.13-7.08 (m, 2H), 2.99 (t, J = 7.5 Hz, 2H), 2.79 (t, J = 7.1 Hz, 2H).

^{13 C NMR (75 MHz, CDCl} 3) δ201.5, 140.5, 138.2, 129.2, 128.6, 127.1, 125.4, 45.3, 28.1, 21.4.

Step 2. 2-Amino-5- (3- Methylbenzyl ) -3,7- Dihydro -4H- Pyrrolo [2,3-d] pyrimidin- Preparation of pyrimidin-4-one

3- (m-Tolyl) propane (912 mg, 6.15 mmol), bromotrimethylsilane (1.22 mL, 9.23 mmol) prepared in the above step (1) and 2-bromo-3- (m-tolyl) propanal (1.40 g, qaunt. Yield) was obtained from 4- (4-amino-6-hydroxypyridin- The objective compound (648 mg, 41% yield) was obtained from the above-prepared sodium salt (6.15 mmol), sodium acetate (1.01 mg, 12.30 mmol) and 2-bromo-3- (m- ).

mp 286.1-287.4 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ10.69 (s, 1H), 10.12 (s, 1H), 7.13-7.08 (m, 4H), 6.93 (d, J = 6.7 Hz, 1H), 6.26 (s, 1 H), 6.00 (s, 2 H), 3.89 (s, 2 H), 2.24 (s, 3 H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.8, 152.6, 151.6, 142.6, 137.3, 129.6, 128.3, 126.5, 126.1, 118.2, 114.4, 99.2, 32.1, 21.5.

Step 3. 4- Chloro -5- (3- Methylbenzyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(3-methylbenzyl) -3,7-dihydro-4H-pyrrolo [2,3-d] pyridine prepared in the above step (2) ] Pyrimidin-4-one (563 mg, 2.21 mmol), acetic anhydride (2.5 mL), benzyltriethylammonium chloride (1.01 g, 4.43 mmol), N, N-dimethylaniline (0.56 mL, The desired compound (478 mg, 79% yield) was obtained from porous (V) oxychloride (2.59 mL, 13.28 mmol) and methanolic methanolic 3N HCl (7.4 mL).

mp 232.5-233.6 DEG C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.24 (s, 1H), 7.14 (t, J = 7.3 Hz, 2H), 7.01-6.98 (m, 3H), 6.77 (s, 1H), 6.45 (s, 2 H), 3.99 (s, 2 H), 2.24 (s, 3 H);

^{13 C NMR (75 MHz, DMSO-} d 6) δ159.6, 155.6, 151.3, 141.4, 137.6, 129.4, 128.5, 126.9, 125.9, 121.8, 113.3, 107.8, 31.7, 21.5.

Step 4. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (3-methylbenzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(3-methylbenzyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine prepared in Step (3) in the same manner as in Step (3) (292 mg, 63%) was obtained from 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (254 mg, 1.14 mmol) and potassium carbonate (456 mg, Yield).

mp 203.4-204.5 C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.23 (s, 1H), 7.17 (t, J = 6.7 Hz, 2H), 7.03-7.01 (m, 3H), 6.57 (s, 1H), 5.31 (s 2H), 5.09 (s, 2H), 4.11 (s, 2H), 3.78 (s, 3H), 2.32 (s, 3H), 2.27 (s, 3H), 2.21

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 158.3, 154.03, 153.97, 152.4, 149.4, 140.4, 137.8, 129.4, 128.2, 126.7, 126.0, 125.7, 125.2, 124.5, 114.8, 109.3, 59.9, 47.4, 31.8, 21.4, 13.3, 10.9.

Example  3. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (4- Methylbenzyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 3)

[Compound 3]

[Total reaction formula]

Step 1. Preparation of 3- (p-tolyl) Propanal  Produce

(2.00 g, 9.17 mmol), benzyltriethylammonium chloride (2.09 g, 9.17 mmol), sodium hydrogencarbonate (1.54 g, 18.35 mmol), and allyl chloride in the same manner as in step (1) The objective compound (1.11 g, 81% yield) was obtained from alcohol (1.25 mL, 18.35 mmol) and palladium (II) acetate (494 mg, 2.20 mmol).

^{1 H NMR (300 MHz, CDCl} 3) δ9.82 (s, 1H), 7.49 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 8.0 Hz, 2H), 2.95 (t, J = 7.5 Hz, 2H), 2.77 (t, J = 7.6 Hz, 2H), 2.43 (s, 3H);

^{13 C NMR (75 MHz, CDCl} 3) δ201.7, 137.3, 135.8, 129.3, 128.2, 45.4, 27.7, 21.0.

Step 2. 2-Amino-5- (4- Methylbenzyl ) -3,7- Dihydro -4H- Pyrrolo [2,3-d] pyrimidin- Preparation of pyrimidin-4-one

(1.11 g, 7.46 mmol), bromotrimethylsilane (1.48 mL, 11.19 mmol) prepared in the above step (1) in the same manner as in the step (1) 2-bromo-3- (p-tolyl) propanal (1.69 g, qaunt. Yield) was obtained from 4-diamino-6-hydroxypyrimidine (0.38 mL, (1.22 g, 64% yield) from the above-prepared sodium salt (1.22 g, 7.46 mmol) and sodium acetate (1.22 g, 14.91 mmol) and 2-bromo- ).

mp 294.1-295.1 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ10.67 (s, 1H), 10.11 (s, 1H), 7.16 (d, J = 7.7 Hz, 2H), 7.02 (d, J = 7.6 Hz, 2H ), 6.24 (s, IH), 5.99 (s, 2H), 3.87 (s, 2H), 2.23 (s, 3H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.7, 152.6, 151.5, 139.6, 134.6, 129.0, 128.9, 118.4, 114.2, 99.2, 31.8, 21.1.

Step 3. 4- Chloro -5- (4- Methylbenzyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-methylbenzyl) -3,7-dihydro-4H-pyrrolo [2,3-d] pyridine prepared in the above step (2) (1.06 g, 4.17 mmol), acetic anhydride (5 mL), benzyltriethylammonium chloride (1.90 g, 8.33 mmol), N, N-dimethylaniline The objective compound (243 mg, 21% yield) was obtained from porous (V) oxychloride (4.87 mL, 24.99 mmol) and methanolic methanolic 3N HCl (14 mL).

mp 247.4-248.5 DEG C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.23 (s, 1H), 7.07 (s, 4H), 6.76 (s, 1H), 6.44 (s, 2H), 3.99 (s, 2H), 2.25 (s, 3 H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.6, 155.7, 151.3, 138.4, 135.1, 129.2, 128.7, 121.8, 113.5, 107.8, 31.4, 21.1.

Step 4. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (4-methylbenzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-methylbenzyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine prepared in step (3) in the same manner as in step (3) (152 mg, 58%) was obtained from 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (144 mg, 0.65 mmol) and potassium carbonate (259 mg, 1.87 mmol) Yield).

mp 188.4-189.8 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.23 (s, 1H), 7.10 (s, 4H), 6.57 (s, 1H), 5.31 (s, 2H), 5.03 (s, 2H), 4.11 (s , 2H), 3.76 (s, 3H), 2.33 (s, 3H), 2.27 (s, 3H), 2.21 (s, 3H);

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 158.3, 154.1, 154.0, 152.4, 149.4, 137.5, 135.4, 129.0, 128.5, 126.0, 125.2, 124.4, 115.0, 109.3, 59.9, 47.4, 31.5, 21.0, 13.3, 10.9.

Example  4. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (4-methoxybenzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 4)

[Compound 4]

[Total reaction formula]

Step 1. Preparation of 3- (4- Methoxyphenyl ) Propanal  Produce

(1.00 g, 6.17 mmol) was dissolved in methanol (12 mL) under a hydrogen atmosphere. Palladium (131 mg, 0.20 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 2 hours. The completion of the reaction was confirmed by TLC (hexane: ethyl acetate = 2: 1). The reaction solution was filtered through a silicate pad with dichloromethane (20 mL x 3) and concentrated under reduced pressure to remove the solvent. The concentrate was separated and purified by column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired compound (619 mg, 66% yield).

^{1 H NMR (300 MHz, CDCl} 3) δ9.83 (s, 1H), 7.15 (d, J = 8.6 Hz, 2H), 6.88 (d, J = 8.6 Hz, 2H), 3.81 (s, 3H), 2.94 (t, J = 7.3 Hz, 2H), 2.76 (t, J = 7.3 Hz, 2H);

^{13 C NMR (75 MHz, CDCl} 3) δ201.8, 158.1, 132.4, 129.3, 114.0, 103.8, 55.3, 45.5, 27.3.

Step 2. 2-Amino-5- (4-methoxybenzyl) -3,7- Dihydro -4H- Pyrrolo [2,3-d] pyrimidine ≪ / RTI >

3- (4-methoxyphenyl) propane (1.25 g, 7.59 mmol), bromotrimethylsilane (1.50 mL, 11.38 mmol) prepared in the above step (1) 2-bromo-3- (4-methoxyphenyl) propanal (1.85 g, qaunt. Yield) was obtained from 4- (4-methoxyphenyl) Bromo-3- (4-methoxyphenyl) propane (1.85 g, 7.59 mmol), which was prepared in the above-mentioned experiment, from the compound (957 mg, 7.59 mmol) and sodium acetate 1.45 g, 71% yield).

mp 268.3-270.0 C;

^{+ isomer 1 H NMR (300 MHz} , DMSO- d 6) δ10.72 (s, 1H), 10.66 (s, 1H), 10.13 (s, 1H), 10.11 (s, 1H), 7.48 (s, 1H) , 7.26 (d, J = 8.4 Hz, 1H), 7.19 (d, J = 8.5 Hz, 2H), 6.97 (d, J = 8.5 Hz, 1H), 6.79 (d, J = 8.5 Hz, 2H), 6.35 (s, 3H), 3.78 (s, 3H), 3.78 (s, 2H), 3.99 (s, 3H);

^{13 C NMR (75 MHz, DMSO-} d 6) δ159.73, 159.70, 157.7, 153.8, 152.7, 152.6, 151.7, 151.6, 136.8, 134.7, 133.1, 129.9, 129.3, 118.7, 117.9, 114.4, 114.1, 113.9, 112.9, 110.5, 99.1, 99.0, 56.6, 55.4, 31.3, 30.9.

Step 3. 4- Chloro -5- (4-methoxybenzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-methoxybenzyl) -3,7-dihydro-4H-pyrrolo [2,3-c] pyridine prepared in step (2) in the same manner as in step (2) (1.14 g, 4.21 mmol), acetic anhydride (4.7 mL), benzyltriethylammonium chloride (1.92 g, 8.41 mmol), N, N-dimethylaniline (1.07 mL, 8.41 mmol) The objective compound (903 mg, 74% yield) was obtained from phosphorus (V) oxychloride (4.92 mL, 25.24 mmol) and methanolic methanolic 3N HCl (14 mL).

mp 237.5-238.7 DEG C;

^{+ isomer 1 H NMR (300 MHz} , DMSO- d 6) δ11.28 (s, 1H), 7.37 (s, 1H), 7.18-7.10 (m, 2H), 7.00 (d, J = 8.4 Hz, 1H) , 6.84 (s, 2H), 6.74 (s, 1H), 6.47 (s, 2H), 3.97 (s, 3H), 3.79 (s, 3H), 3.70

^{13 C NMR (75 MHz, DMSO-} d 6) δ159.7, 159.6, 157.9, 155.7, 154.0, 151.3, 151.2, 135.4, 133.3, 132.9, 129.8, 129.2, 122.0, 121.6, 114.1, 113.8, 113.1, 113.0, 110.7, 107.7, 107.6, 56.6, 55.4, 31.0, 30.5.

Step 4. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (4-methoxybenzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-methoxybenzyl) -7H-pyrrolo [2,3-d] pyrimidine-2-carbaldehyde prepared in the above step (3) in the same manner as in step (3) (214 mg, 47%) was obtained from 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (242 mg, 1.09 mmol) and potassium carbonate (431 mg, % Yield).

mp 191.6-192.8 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.22 (s, 1H), 7.13 (d, J = 8.4 Hz, 2H), 6.82 (d, J = 8.4 Hz, 2H), 6.54 (s, 1H), 2H), 5.07 (s, 2H), 4.08 (s, 2H), 3.80 (s, 3H), 3.76 (s, 3H), 2.27 (s, 3H), 2.21

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 158.3, 157.9, 154.1, 154.0, 152.4, 149.5, 149.4, 132.6, 129.6, 126.0, 125.2, 124.3, 115.2, 113.7, 59.9, 55.3, 47.4, 31.1, 13.3, 10.9.

Example  5. 5- (4- Bromo -3-methoxybenzyl) -4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 5)

[Compound 5]

[Total reaction formula]

Step 1. Preparation of 3- (3- Methoxyphenyl ) Propanal  Produce

3-iodoanisole (1.02 mL, 8.55 mmol), benzyltriethylammonium chloride (1.95 g, 8.55 mmol), sodium hydrogen carbonate (1.44 g, 17.09 mmol) The objective compound (904 mg, 64% yield) was obtained from allyl alcohol (1.16 mL, 17.09 mmol) and palladium (II) acetate (1.16 mL, 17.09 mmol).

^{1 H NMR (300 MHz, CDCl} 3) δ9.72 (s, 1H), 7.19 (t, J = 8.1 Hz, 1H), 6.77-6.73 (m, 3H), 3.74 (s, 3H), 2.88 (t, J = 7.4 Hz, 2H), 2.69 (t, J = 7.4 Hz, 2H);

^{13 C NMR (75 MHz, CDCl} 3) δ201.4, 159.8, 142.2, 129.6, 120.6, 114.2, 111.5, 55.0, 45.0, 28.1.

Step 2. 2-Amino-5- (4- Bromo -3-methoxybenzyl) -3,7- Dihydro -4H-pyrrolo [ 2,3-d] pyrimidine -4-one

3- (3-methoxyphenyl) propane (904 mg, 5.50 mmol), bromotrimethylsilane (1.09 mL, 8.26 mmol) prepared in the above step (1) Bromo-3- methoxyphenyl) propane (1.34 g, 75% yield) was obtained from 2-bromo-3- (694 mg, 5.50 mmol), sodium acetate (903 mg, 11.01 mmol) and 2-bromo-3- (4-bromo-3-methoxyphenyl) propanel 1.34 g, 5.50 mmol), the desired compound (1.50 g, 75% yield) was obtained.

mp 288.1-289.3 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ10.76 (s, 1H), 10.21 (s, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.03 (d, J = 2.9 Hz, 1H ), 6.71 (dd, J = 8.7 Hz, 3.0 Hz, 1H), 6.28 (s, 1H), 6.04 (s, 2H), 3.99 (s, 2H), 3.67 (s, 3H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.7, 159.1, 152.7, 151.7, 142.5, 133.1, 117.3, 115.8, 114.9, 114.5, 113.6, 99.2, 55.6, 32.5.

Step 3. Preparation of 5- (4- Bromo -3-methoxybenzyl) -4- Chloro -7H- Pyrrolo [2,3-d] pyrimidine -2-amine

(4-bromo-3-methoxybenzyl) -3,7-dihydro-4H-pyrrolo [2,3-d] pyrimidin- (1.20 g, 3.44 mmol), acetic anhydride (4 mL), benzyltriethylammonium chloride (1.57 g, 6.87 mmol) and N, N-dimethylaniline (0.87 mL , 6.87 mmol), phosphorus (V) oxychloride (4.02 mL, 20.62 mmol) and methanolic methanolic 3N HCl (12 mL).

mp 225.6-226.6 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.29 (s, 1H), 7.51 (d, J = 8.7 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 6.66 (s, 1H ), 6.64 (s, IH), 6.49 (s, IH), 4.07 (s, 2H), 3.65 (s, 3H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.7, 159.2, 155.7, 155.6, 151.3, 141.5, 133.4, 122.2, 122.0, 117.0, 114.6, 113.7, 111.3, 107.7, 55.7, 32.5.

Step 4. Preparation of 5- (4- Bromo -3-methoxybenzyl) -4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

3-methoxybenzyl) -4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-ylamine obtained in step (3) (300 mg, 0.82 mmol) and 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (189 mg, 0.85 mmol) and potassium carbonate (338 mg, (152 mg, 27% yield).

mp 232.3-233.3 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.21 (s, 1H), 7.45 (d, J = 9.4 Hz, 1H), 6.64 (s, 2H), 6.55 (s, 1H), 5.32 (s, 2H ), 5.01 (s, 2H), 4.19 (s, 2H), 3.76 (s, 3H), 3.71 (s, 3H), 2.27 (s, 3H), 2.21

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 159.0, 158.4, 154.1, 153.9, 152.5, 149.4, 141.1, 133.1, 126.0, 125.2, 124.9, 116.4, 115.0, 113.1, 112.7, 109.3, 59.9, 55.3, 47.5, 32.5, 13.3, 10.9.

Example  6. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (3- ( Trifluoromethyl ) Benzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 6)

[Compound 6]

[Total reaction formula]

Step 1. Preparation of 3- (3- ( Trifluoromethyl ) Phenyl ) Propanal  Produce

3-iodobenzotrifluoride (1.06 mL, 7.35 mmol), benzyltriethylammonium chloride (1.68 g, 7.35 mmol) and sodium bicarbonate (1.24 g, 14.71 mmol) were obtained in the same manner as in step (1) ), The objective compound (561 mg, 38% yield) was obtained from allyl alcohol (1.00 mL, 14.71 mmol) and palladium (II) acetate (396 mg, 1.77 mmol).

^{1 H NMR (300 MHz, CDCl} 3) δ9.85 (s, 1H), 7.37-7.28 (m, 1H), 7.16 (d, J = 7.6 Hz, 1H), 7.08 (s, 2H), 3.00 (t , J = 7.4 Hz, 2H), 2.83 (t, J = 7.1 Hz, 2H);

¹³ C NMR (75 MHz, CDCl ₃ )? 200.7, 149.4, 142.8, 129.9, 126.8, 120.9, 118.7, 44.9, 27.7.

Step 2. 2-Amino-5- (3- ( Trifluoromethyl ) Benzyl) -3,7- Dihydro -4H-pyrrolo [ 2,3-d] pyrimidine -4-one

3- (3- (trifluoromethyl) phenyl) propane (1.14 g, 5.65 mmol), bromotrimethylsilane (1.12 g, 5.65 mmol) prepared in the above step (1) bromo-3- (4-chlorophenyl) propanal (1.59 g, qaunt. yield) was obtained from 4-diamino-6-hydroxynaphthalene Bromo-3- (4-chlorophenyl) propanal (1.59 g, 5.65 mmol) prepared in the above-mentioned experiment was dissolved in methyl ethyl ketone (525 mg, 30% yield).

mp 264.9-266.2 DEG C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ10.76 (s, 1H), 10.20 (s, 1H), 7.39-7.31 (m, 3H), 7.11 (d, J = 6.7 Hz, 1H), 6.40 (s, 1 H), 6.04 (s, 2 H), 3.97 (s, 2 H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.7, 152.7, 151.7, 148.8, 145.8, 130.3, 128.1, 125.6, 122.3, 121.3, 118.9, 118.3, 117.1, 115.5, 114.6, 99.0, 31.8.

Step 3. 4- Chloro -5- (3- ( Trifluoromethyl ) Benzyl) -7H- Pyrrolo [2,3-d] pyrimidin- Pyrimidin-2- Amine  Produce

(Trifluoromethyl) benzyl) -3,7-dihydro-4H-pyrrolo [2,3-c] pyridine prepared in the above step (2) in the same manner as in step (2) N, N-dimethylaniline (0.43 mL, 3.40 mmol), benzyltriethylammonium chloride (774 mg, 3.40 mmol) The objective compound (324 mg, 56% yield) was obtained from methanolic methanolic 3N HCl (6 mL) and phosphorus (V) oxychloride (2.0 mL, 10.19 mmol).

mp 203.6-204.5 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.31 (s, 1H), 7.40 (t, J = 8.2 Hz, 1H), 7.23 (d, J = 7.8 Hz, 2H), 7.17 (s, 1H ), 6.89 (s, 1 H), 6.48 (s, 2 H), 4.10 (s, 2 H);

^{13 C NMR (75 MHz, DMSO-} d 6) δ159.7, 155.7, 151.2, 148.8, 144.6, 130.5, 127.9, 122.2, 121.1, 118.7, 112.2, 107.6, 31.4.

Step 4. 4- Chloro -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl ) -5- (3- ( triple Methyl) benzyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

Pyrrole [2,3-d] pyrimidin-2-ylamine obtained in the above step (3) in the same manner as in step (3) Amine (200 mg, 0.61 mmol) and 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (141 mg, 0.64 mmol) and potassium carbonate (254 mg, 1.84 mmol) 208 mg, 71% yield).

187.4-188.2 DEG C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.22 (s, 1H), 7.30 (t, J = 7.6 Hz, 1H), 7.15 (d, J = 7.5 Hz, 1H), 7.07 (s, 1H), 2H), 3.76 (s, 3H), 2.27 (s, 3H), 2.22 (s, 2H) (s, 3 H);

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 158.4, 154.1, 153.8, 152.4, 149.5, 149.3, 143.0, 129.5, 127.0, 126.1, 125.2, 124.6, 121.1, 118.4, 113.5, 109.1, 59.9, 47.4, 31.7, 13.3, 10.9.

Example  7. 4- Chloro -5- (4- Chlorobenzyl ) -7 - ((4- Methoxy -3,5-dimethylpyridin-2-yl) methyl) -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 7)

[Compound 7]

[Total reaction formula]

Step 1. Preparation of 3- (4- Chlorophenyl ) Propanal  Produce

(2.00 g, 8.39 mmol), benzyltriethylammonium chloride (1.91 g, 8.39 mmol), sodium hydrogencarbonate (1.41 g, 16.78 mmol), and diisopropylethylamine in the same manner as in the step (1) (695 mg, 49% yield) was obtained from allyl alcohol (1.14 mL, 16.78 mmol) and palladium (II) acetate (452 mg, 2.01 mmol).

^{1 H NMR (300 MHz, CDCl} 3) δ9.84 (s, 1H), 7.29 (d, J = 8.4 Hz, 2H), 7.16 (d, J = 8.3 Hz, 2H), 2.96 (t, J = 7.4 Hz, < / RTI > 2H), 2.80 (t, J = 6.9 Hz, 2H);

^{13 C NMR (75 MHz, CDCl} 3) δ201.0, 138.9, 132.1, 129.7, 128.7, 45.1, 27.4.

Step 2. 2-Amino-5- (4- Chlorobenzyl ) -3,7- Dihydro -4H- Pyrrolo [2,3-d] pyrimidine ≪ / RTI >

3- (4-chlorophenyl) propane (695 mg, 4.12 mmol), bromotrimethylsilane (0.82 mL, 6.19 mmol) prepared in the above step (1) 2-bromo-3- (4-chlorophenyl) propane (1.02 g, qaunt. Yield) was obtained from 4- bromo-4-methoxybenzaldehyde (0.21 mL, 4.12 mmol) bromo-3- (4-chlorophenyl) propane (1.02 g, 4.13 mmol), which was prepared in the above-mentioned experiment, and the desired compound (496 mg, 44% yield).

mp 289.0-290.0 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ10.73 (s, 1H), 10.13 (s, 1H), 7.32-7.25 (m, 4H), 6.33 (s, 1H), 6.02 (s, 2H) , ≪ / RTI > 3.91 (s, 2H);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 159.7, 152.7, 151.7, 141.9, 130.8, 130.5, 128.3, 117.5, 114.5, 99.1, 31.5.

Step 3. 4- Chloro -5- (4- Chlorobenzyl ) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-chlorobenzyl) -3,7-dihydro-4H-pyrrolo [2,3-d] pyridine prepared in the above step (2) ] Pyrimidin-4-one (399 mg, 1.45 mmol), acetic anhydride (2.5 mL), benzyltriethylammonium chloride (661 mg, 2.90 mmol), N, N-dimethylaniline The desired compound (236 mg, 55% yield) was obtained from porous (V) oxychloride (1.70 mL, 8.71 mmol) and methanolic methanolic 3N HCl (5 mL).

mp 272.3-273.3 [deg.] C;

^{1 H NMR (300 MHz, DMSO-} d 6) δ11.29 (s, 1H), 7.31 (d, J = 7.9 Hz, 2H), 7.21 (d, J = 8.1 Hz, 2H), 6.84 (s, 1H ), 6.47 (s, 2 H), 4.03 (s, 2 H);

^{13 C NMR (75 MHz, DMSO-} d 6) δ159.7, 155.7, 151.2, 140.7, 130.8, 130.6, 128.6, 122.1, 112.6, 107.6, 31.1.

Step 4. 4- Chloro -5- (4- Chlorobenzyl ) -7 - ((4- Methoxy -3,5- Dimethylpyridine Yl) methyl) -7H- Pyrrolo [2,3-d] pyrimidine -2- Amine  Produce

(4-chlorobenzyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine prepared in Step (3) in the same manner as in Step (3) (129 mg, 43%) was obtained from 2-chloromethyl-4-methoxy-3,5-dimethylpyridine (156 mg, 0.70 mmol) and potassium carbonate (279 mg, Yield).

mp 178.0-179.2 DEG C;

^{1 H NMR (300 MHz, CDCl} 3) δ8.22 (s, 1H), 7.24 (d, J = 8.3 Hz, 2H), 7.14 (d, J = 8.3 Hz, 2H), 6.58 (s, 1H), 3H), 2.28 (s, 3H), 2.23 (s, 3H);

^{13 C NMR (75 MHz, CDCl} 3) δ164.3, 158.4, 154.1, 153.8, 152.4, 149.5, 139.1, 131.7, 130.0, 128.4, 126.0, 125.2, 124.6, 114.1, 109.1, 59.9, 47.3, 31.4, 13.3, 10.9.

Example  8. 5,7- Dibenzyl -4- Chloro -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 8)

[Compound 8]

[Total reaction formula]

Benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (0.14 mL, (239 mg, 59% yield) was obtained from benzyl chloride (0.14 mL, 1.21 mmol) and potassium carbonate (481 mg, 3.48 mmol).

mp 150.0-150.9 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ7.33 (t, J = 7.5 Hz, 6H), 7.25 (d, J = 7.1 Hz, 2H), 7.18 (d, J = 6.8 Hz, 2H), 6.46 ( s, 1 H), 5.23 (s, 2 H), 5.07 (s, 2 H), 4.19 (s, 2 H);

^{13 C NMR (75 MHz, CDCl} 3) δ158.5, 154.3, 152.6, 140.3, 137.0, 128.8, 128.7, 128.4, 127.8, 127.3, 126.1, 124.3, 115.2, 109.4, 47.6, 32.1.

Example  9. 7- ( Benzo [d] [1,3] dioxole -5- Yl methyl ) -5-benzyl-4- Chloro -7H- Pyrrolo [2,3-d] pyrimidine -2-amine (Compound 9)

[Compound 9]

[Total reaction formula]

Step 1. 5- ( Chloromethyl ) Benzo [d] [1, 3] Dioxol  Produce

Piperenyl alcohol (2.00 g, 13.15 mmol) and triethylamine (1.94 mL, 13.93 mmol) were dissolved in dichloromethane (44 mL) under an argon atmosphere and then cooled to 0 ° C. Oleyl (1.44 mL, 19.72 mmol) was added dropwise. The reaction solution was stirred at room temperature for 2 hours, and the completion of the reaction was confirmed by TLC (hexane: ethyl acetate = 2: 1). The reaction solution was cooled to 0 ° C and washed with distilled water (65 mL), 2N aqueous hydrochloric acid chloride solution (10 mL) and saturated sodium bicarbonate solution (40 mL), and the organic solvent layer extracted with dichloromethane (15 mL x 3) The organic solvent layer was washed with saturated solution (25 mL), dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to remove the solvent. The concentrate was separated and purified by column chromatography (hexane: ethyl acetate = 3: 1) to obtain the title compound (2.11 g, 94% yield).

^{1 H NMR (300 MHz, CDCl} 3) δ6.93 (s, 1H), 6.89 (d, J = 7.9 Hz, 1H), 6.82 (d, J = 7.9Hz, 1H), 5.99 (s, 2H), 4.57 (s, 2H);

¹³ C NMR (75 MHz, CDCl ₃ )? 148.0, 147.9, 131.4, 122.4, 109.2, 108.3, 101.4, 46.7.

Step 2. 7- ( Benzo [d] [1,3] dioxole -5- Yl methyl ) -5-benzyl-4- Chloro -7H-pyrrolo [ 2,3-d] pyrimidine -2- Amine  Produce

Benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (300 mg, 1.16 mmol) prepared in the step (2) (274 mg, 60%) was obtained from 5- (chloromethyl) benzo [d] [1,3] dioxole (206 mg, 1.21 mmol) and potassium carbonate (481 mg, 3.48 mmol) Yield).

mp 175.8-177.1 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ7.34-7.24 (m, 5H), 6.76 (s, 1H), 6.70 (s, 2H), 6.45 (s, 1H), 5.96 (s, 2H), 5.10 (s, 2 H), 5.07 (s, 2 H), 4.17 (s, 2 H);

¹³ C NMR (75 MHz, CDCl ₃ )? 158.5, 154.2, 152.6, 148.1, 147.3, 140.3, 130.8, 128.7, 128.4, 126.1, 124.0, 120.9, 115.2, 109.4, 108.4, 108.1, 101.2, 47.4, 32.1.

Example  10. Preparation of 5-benzyl-7 - ((6- Bromobenzo [d] [1,3] dioxole -5 days) methyl )-4- Chloro -7H-P 2,3-d] pyrimidine 2-amine (Compound j)

[Compound 10]

[Total reaction formula]

Step 1. 5- Bromo -6- ( Chloromethyl ) Benzo [d] [1, 3] Dioxol  Produce

(Chloromethyl) benzo [d] [1,3] dioxole (2.29 g, 13.42 mmol) prepared in step (1) of Example 9 was dissolved in acetonitrile -Bromosuccinimide (2.39 g, 16.11 mmol) was added. The reaction solution was stirred at room temperature for 13 hours, and the completion of the reaction was confirmed by TLC (hexane: ethyl acetate = 2: 1). The reaction mixture was poured into water and the organic solvent layer extracted with dichloromethane (10 mL x 3) was washed again with a saturated sodium chloride solution (50 mL). The organic solvent layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to remove the solvent Respectively. The concentrate was separated and purified by column chromatography (hexane: ethyl acetate = 5: 1) to obtain the desired compound (3.32 g, 99% yield).

^{1 H NMR (300 MHz, CDCl} 3) δ7.02 (s, 1H), 6.94 (s, 1H), 6.01 (s, 2H), 4.64 (s, 2H);

¹³ C NMR (75 MHz, CDCl ₃ )? 148.7, 147.6, 129.7, 115.2, 112.9, 110.4, 102.2, 46.6.

Step 2. Preparation of 5-benzyl-7 - ((6- Bromobenzo [d] [1,3] dioxole -5 days) methyl )-4- Chloro -7H- blood Lolo [ 2,3-d] pyrimidine -2- Amine  Produce

Benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (300 mg, 1.16 mmol) prepared in the step (2) (301 mg, 1.21 mmol) and potassium carbonate (481 mg, 3.48 mmol), which was prepared in step (1) above, to give the title compound 304 mg, 56% yield).

mp 210.3-210.8 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ7.35-7.22 (m, 5H), 7.05 (s, 1H), 6.53 (s, 1H), 6.45 (s, 1H), 5.98 (s, 2H), 5.21 (s, 2 H), 5.01 (s, 2 H), 4.19 (s, 2 H);

^{13 C NMR (75 MHz, CDCl} 3) δ158.5, 154.3, 152.7, 148.1, 147.8, 140.2, 129.4, 128.7, 128.4, 126.2, 124.3, 115.3, 113.3, 112.8, 109.3, 109.0, 101.9, 47.5, 32.1.

Example  11. 5-Benzyl- Chloro -7 - ((6- Iodobenzo [d] [1,3] dioxole -5 days) methyl ) -7H-P 2,3-d] pyrimidine 2-amine (Compound 11)

[Compound 11]

[Total reaction formula]

Step 1. (6- Iodobenzo [d] [1,3] dioxole -5-yl) methanol

Pyrrolyl alcohol (2.50 g, 16.43 mmol) and silver trifluoroacetate (4.36 g, 19.72 mmol) were dissolved in dichloromethane (51 mL) under an argon atmosphere and then iodine (5.00 g , 19.72 mmol) was slowly added dropwise at 0 ° C. The reaction solution was stirred at room temperature for 14 hours, and the completion of the reaction was confirmed by TLC (hexane: ethyl acetate = 2: 1). After the reaction, the resulting solid is filtered through silyl chloride with dichloromethane and concentrated under reduced pressure to remove the solvent. The concentrate was extracted with dichloromethane (20 mL x 3), and the extracted organic solvent layer was washed with a saturated sodium metabisulfate solution (20 mL) and a saturated sodium chloride solution (50 mL). The organic solvent layer was dried over anhydrous magnesium sulfate, The solvent was removed by concentration under reduced pressure. The concentrate was separated and purified by column chromatography (hexane: ethyl acetate = 6: 1) to obtain the desired compound (4.57 g, quant.

^{1 H NMR (300 MHz, CDCl} 3) δ7.27 (s, 1H), 7.02 (s, 1H), 6.01 (s, 2H), 5.34 (s, 1 H), 4.62 (s, 2 H);

¹³ C NMR (75 MHz, CDCl ₃ )? 148.7, 147.9, 136.3, 118.6, 109.1, 101.7, 85.4, 69.3.

Step 2. 5- ( Chloromethyl ) -6- Iodobenzo [d] [l, 3] dioxole  Produce

(6-Iodobenzo [d] [1,3] dioxol-5-yl) methanol (300 mg, 1.16 mmol) prepared in the above step (1) in the same manner as in step (1) The desired compound (4.87 g, quant. Yield) was obtained from triethylamine (2.43 mL, 17.42 mmol) and thionyl chloride (1.80 mL, 24.65 mmol).

^{1 H NMR (300 MHz, CDCl} 3) δ7.30 (s, 1H), 7.01 (s, 1H), 6.03 (s, 2H), 4.66 (s, 2H);

¹³ C NMR (75 MHz, CDCl ₃ )? 148.73, 148.67, 133.2, 118.9, 110.1, 102.0, 88.2, 51.4.

Step 3. 5-Benzyl-4- Chloro -7 - ((6- Iodobenzo [d] [1,3] dioxole -5 days) methyl ) -7H-pyrrolo [ 2,3-d] pyrimidine -2- Amine  Produce

Benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine (205 mg, 0.79 mmol) prepared in the step (2) (245 mg, 0.83 mmol) and potassium carbonate (329 mg, 2.38 mmol), which was prepared in step (1), to obtain the title compound 220 mg, 53% yield).

mp 189.5-190.3 [deg.] C;

^{1 H NMR (300 MHz, CDCl} 3) δ7.35-7.22 (m, 6H), 6.51 (s, 1H), 6.41 (s, 1H), 5.97 (s, 2H), 5.16 (s, 2H), 5.02 (s, 2 H), 4.19 (s, 2 H);

^{13 C NMR (75 MHz, CDCl} 3) δ158.5, 154.3, 152.7, 148.9, 148.1, 140.2, 132.6, 128.7, 128.4, 126.2, 124.2, 118.7, 115.4, 109.3, 108.7, 101.8, 85.8, 52.3, 32.1.

Experimental Example  1. Anticancer effect of cancer cell inhibitor ( MCF -7, HCT116 , SK - BR -3)

Cell lines used in the MTT assay were MCF-7 (human breast cancer), SK-BR-3 (human breast cancer) and HCT116 (human colon carcinoma cancer). The culture medium was RPMI 1640 medium containing 10% fetal bovine serum and incubated at 37 ° C in a constant temperature and humidity 5% CO ₂ incubator. Cells were cultured in a 96-well microplate for 3 x 10 ³ cells per well. After culturing for 24 h in a CO ₂ incubator, media was removed and 100 μl of each diluted inhibitor solution was added for 72 h Lt; / RTI > 15 μl of MTT solution (Promega, CellTiter 96) was added and incubated for 4 hours. Stop solution (Promega, CellTiter 96) to dissolve blue formazan. After standing for 18 hours or longer, the absorbance was measured at 570 nm to obtain GI ₅₀ (concentration at which cancer cells were inhibited by 50%). The inhibitory activity against the cancer cells and the GI ₅₀ were measured at 72 hours after the compounds 1 to 11 were treated or not treated at a concentration or concentration of 10 μM, respectively, and the results are shown in Table 1 for each compound added .

The following Table 1 quantifies the activity of cancer cells at the concentration of the compound, with the cancer cell activity of the compound not treated (negative control group) as 100%. A positive control was BIIB021 (6-chloro-9 - [(4-methoxy-3,5-dimethylpyridin-2- -17-demethoxygeldanamycin, anti-cancer substance) was used. Both BIIB021 and 17-AAG are clinical anticancer substances, BIIB021 was synthesized directly in the laboratory and 17-AAG was purchased from LC Laboratroies®.

compound MCF -7 SK - BR -3 HCT116 % Inhibition
10 μ M GI ₅₀ ,
μ M % Inhibition
10 μ M GI ₅₀ ,
μ M % Inhibition
10 μ M GI ₅₀ ,
μ M One 85.61 0.35
± 0.003 69.9 0.69
± 0.021 88.52 0.52
± 0.031 2 86.53 0.69
± 0.02 87.15 0.83
± 0.11 94.18 0.87
± 0.01 3 83.20 1.09
± 0.09 81.04 1.48
± 0.01 71.33 1.72
± 0.16 4 80.06 1.35
± 0.08 80.89 1.49
± 0.07 69.43 1.95
± 0.24 5 62.01 4.44
± 0.10 78.70 3.94
± 0.09 80.24 3.65
± 0.21 6 79.09 4.95
± 0.77 21.58 2.55
± 0.57 89.77 1.51
± 0.26 7 87.97 1.05
± 0.04 72.59 1.53
± 0.23 90.09 1.08
± 0.09 8 26.4 - 80.54 - 30.57 - 9 54.03 - 4.32 - 48.75 - 10 69.94 20.80
± 2.40 40.92 8.34
± 0.67 70.41 5.15
± 0.15 11 74.04 6.40
± 0.14 63.8 7.55
± 0.27 74.49 5.05
± 0.10 BIIB021 84.81 0.392
± 0.03 86.20 0.227
± 0.015 71.94 0.347
± 0.023 17- AAG 79.74 0.498
± 0.06 88.89 0.205
± 0.009 74.44 0.140
± 0.012

As shown in Table 1, when the cancer cells (%) of the negative control (control, DMSO-treated cells) were set to 100%, the inhibitory effect of the compounds according to Examples 1 to 11 of the present invention on cancer cells was confirmed by MCF- 7 cancer cells at a concentration of 10 [mu] M from at least about 26.4% up to 87.97%. In HCT116 cancer cells, the inhibition was reduced from at least about 30.57% to 94.18%, and in SK-BR-3 cancer cells at least about 4.32% to 87.15%. GI ₅₀ was measured for compounds exhibiting high% inhibition. As a result, Compound 1 was found to have a similar inhibitory activity to BIIB021 at a concentration of about 0.35 ± 0.003 μM in MCF-7 cancer cells and higher than the positive control 17-AAG, BR-3 and HCT116 showed the best activities in the compounds synthesized at about 0.69 ± 0.021 and 0.52 ± 0.031 μM. Compound 2 also showed excellent cancer cell inhibitory activity at the concentrations of about 0.69 ± 0.02, 0.83 ± 0.11 and 0.87 ± 0.01 μM in cancer cells of MCF-7, SK-BR-3 and HCT116.

Experimental Example  2. Cytotoxicity measurement in normal cells

Compound 1, which had a good cancer cell inhibitory activity, was treated with 10 uM of HFF-1 (Human Fibroblast) normal cell line (ATCC®, SCRC-1041) in the same manner as in Experimental Example 1, The% inhibitory activity against normal cells was measured at one time and the results are shown in Table 2 below.

As a result of cytotoxicity measurement, Compound 1 having the most excellent anticancer activity showed 46.74% inhibition effect at 10 uM concentration on normal cells, which is lower than that of the control substances (BIIB021 and 17-AAG) Compared with 28 times less toxic.

compound HFF-1 % Inhibition
10 uM One 46.74 BIIB021 51.28 17-AAG 65.49

Therefore, the compound prepared according to the present invention showed a low toxicity to normal cells and an excellent effect of selectively inhibiting cancer cells, and it was confirmed that even when using an excess amount exceeding the effective dose, the risk is very low and safe to use.

Experimental Example  3. MCF -7 < / RTI > Hsp70  Measure induction effect

The compound 1 of the present invention was tested for the degradation of the Hsp90 client proteins EGFR, Her2, Met and Akt in the MCF-7 medium, and the inhibition of Hsp90 was examined by observing the induction of Hsp70.

Compounds of Example 1 were treated with MCF-7 cells at 30, 20, 10, 5, 1, and 0 mM for 24 hours. Expression levels of the client proteins, Hsp90, and Hsp70 were measured by Western blotting. MCF-7 cells were inoculated into 60 mm culture dishes at a concentration of 1 × 10 ⁶ cells / dish, and cells were adhered to the floor for one day. Compounds were cultured for 24 hrs after treatment with different concentrations of cells. Cells were treated with 1% DMSO or gelatinamycin (1 mM, Enzo Life Sciences) for 24 hours as a control. Cells were extracted with cold lysis buffer (25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% NP40, 1% sodium deoxycholate, 0.1% SDS) and 20 mg lysates were separated by SDS-PAGE. After transferring to a PVDF membrane (Bio-Rad), the membranes were blotted onto TBST with 5% skim milk and incubated with antibodies (purchased from Cell Signaling Technology) against EGFR, Her2, Met, Akt, Hsp90, Hsp70, Lt; / RTI > After binding of the second antibody linked to the enzyme (Horseradish peroxidase), the protein was measured by SuperSignal® West Dura Extended Eating Substrate (Thermo Scientific, USA) and the results are shown in FIG.

As shown in FIG. 1, it was confirmed that the compound of Example 1 decreased the levels of EGFR, Her2, MET, and Akt proteins and maximally decreased the levels of Her2 and MET proteins at the concentration of 5 mM. In addition, it was confirmed that the expression of Hsp70 was induced, and that the expression of Hsp90 was not changed. Thus, the compound of the present invention inhibits the function of Hsp90, thereby decreasing the expression level of the client protein and confirming the anti-cancer effect I could.

Thus, the compounds of the present invention are identified as anticancer compositions as inhibitors of Hsp90 and have demonstrated the possibility of being used as the backbone of the compounds to be developed in the future.

Claims (11)

A pyrrolopyrimidine compound represented by the following formula (1), a pharmaceutically acceptable salt or hydrate thereof:
[Chemical Formula 1]

In Formula 1,
R < ¹ > is one or more substituents each independently selected from the group consisting of H; Goat; bromine; Trifluoromethyl; C ₁ -C ₃ alkyl or C ₁ -C ₃ alkoxy group,
R ² , R ³ , R ⁴ and R ⁵ are each independently H; A C ₁ -C ₃ alkyl group; A C ₁ -C ₃ alkoxy group or a dialkoxy connected by a C ₁ -C ₃ chain,
X is carbon or nitrogen,
When X is carbon, X is H; bromine; ≪ / RTI > and iodine. delete The method according to claim 1,
A pyrrolopyrimidine compound selected from the group consisting of the following compounds, their pharmaceutically acceptable salts or hydrates:
5-Benzyl-4-chloro-7 - ((4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine;
5-Benzyl-4-chloro-7 - ((4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methylbenzyl) -7H- pyrrolo [2,3- d] pyrimidine- 2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methoxybenzyl) -7H-pyrrolo [2,3- d] pyrimidine Amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3 -d] pyrimidin-2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (3- (trifluoromethyl) benzyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amine;
Pyrrolo [2,3-d] pyrimidin-2-ylmethyl) -7- (4-methoxy- 2-amine;
5,7-Dibenzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine;
7- (benzo [d] [1,3] dioxol-5-ylmethyl) -5-benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine;
7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) -quinolin- - amine; And
Yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) - Amines.
The method according to claim 1,
The pharmaceutically acceptable salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, Which is a salt of an inorganic or organic acid selected from the group consisting of ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid and toluenesulfonic acid,
A pyrrolopyrimidine compound, a pharmaceutically acceptable salt thereof, or a hydrate thereof.
A pyrrolopyrimidine compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof,
Pharmaceutical composition for preventing or treating cancer:
[Chemical Formula 1]

In Formula 1,
R < ¹ > is one or more substituents each independently selected from the group consisting of H; Goat; bromine; Trifluoromethyl; C ₁ -C ₃ alkyl or C ₁ -C ₃ alkoxy group,
R ² , R ³ , R ⁴ and R ⁵ are each independently H; A C ₁ -C ₃ alkyl group; A C ₁ -C ₃ alkoxy group or a dialkoxy connected by a C ₁ -C ₃ chain,
X is carbon or nitrogen,
When X is carbon, X is H; bromine; ≪ / RTI > and iodine. delete 6. The method of claim 5,
A pharmaceutical composition for preventing or treating cancer comprising as an active ingredient a pyrrolopyrimidine compound selected from the group consisting of the following compounds, a pharmaceutically acceptable salt thereof or a hydrate thereof:
5-Benzyl-4-chloro-7 - ((4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine;
5-Benzyl-4-chloro-7 - ((4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methylbenzyl) -7H- pyrrolo [2,3- d] pyrimidine- 2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (4-methoxybenzyl) -7H-pyrrolo [2,3- d] pyrimidine Amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2,3 -d] pyrimidin-2-amine;
(4-methoxy-3,5-dimethylpyridin-2-yl) methyl) -5- (3- (trifluoromethyl) benzyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amine;
Pyrrolo [2,3-d] pyrimidin-2-ylmethyl) -7- (4-methoxy- 2-amine;
5,7-Dibenzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine;
7- (benzo [d] [1,3] dioxol-5-ylmethyl) -5-benzyl-4-chloro-7H-pyrrolo [2,3-d] pyrimidin-2-amine;
7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) -quinolin- - amine; And
Yl) methyl) -7H-pyrrolo [2,3-d] pyrimidin-2 (1 H) - Amines.
6. The method of claim 5,
The pharmaceutically acceptable salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, Which is a salt of an inorganic or organic acid selected from the group consisting of ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid and toluenesulfonic acid,
A pharmaceutical composition for preventing or treating cancer.
6. The pharmaceutical composition according to claim 5, wherein the pyrrolopyrimidine compound represented by Formula 1, a pharmaceutically acceptable salt thereof, or a hydrate thereof is contained in an amount of 0.5 to 10% by weight based on the total weight of the total pharmaceutical composition.
A pharmaceutical composition for preventing or treating cancer.
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