Pharmaceutical composition for the prevention, alleviation, or treatment of cancer

Abstract

The compounds according to the present invention exhibit excellent anti-cancer effects against triple-negative breast cancer cell line MDA-MB-231 and liver cancer cell lines, as well as common cancer cell lines, such as A549, SK-OV-3, SK-MEL-2, HCT15, etc., and therefore can be advantageously used as anti-cancer agents.

Description

Technical Field

[0001] Background of the Invention 1. Field of the Invention The present invention relates to a pharmaceutical composition for the prevention, alleviation, or treatment of cancer.

Background Art

[0002] 2. Description of the Related Art Despite the fact that the incidence of cancer is increasing with the development of civilization, the treatment of cancer patients still relies on surgery, radiotherapy, and chemotherapy by administering highly cytotoxic anticancer drugs. However, these treatments are generally limited to patients with early-stage cancer or specific types of cancer and cause various side effects. Therefore, there is a need to develop anticancer drugs that are effective and have fewer side effects while having a safety profile.

[0003] In particular, the breast cancer market is known to be a field where medical needs are not met due to a lack of high efficacy and safety of anticancer drugs.

[0004] Regarding breast cancer treatment, various treatment mechanisms targeting patients with different disease characteristics, such as hormone therapy, chemotherapy, and targeted therapy, have been introduced. However, there is a certain clinical need for treatments with excellent anticancer efficacy and safety. Among these, triple-negative breast cancer (TNBC), which accounts for 16% of all breast cancers, is difficult to treat due to its poor prognosis after treatment, and there are not enough available treatments targeting TNBC (Non-Patent Document 1, Rev Peru Med Exp Salud Publica. Oct-Dec 2013;30(4):649-56).

[0005] Currently, treatments for breast cancer include therapies that inhibit the human epidermal growth factor receptor 2 (hereinafter referred to as "HER2") gene involved in tumor growth, and anti-hormonal drugs are widely used. However, in the case of TNBC, all of the HER2 receptor, estrogen receptor, and progesterone receptor are negative, and thus do not respond to existing anti-cancer drugs. As a result, there is an urgent need for new targeted treatments that can treat safely and efficiently.

[0006] Therefore, the present inventors have studied and completed the pharmaceutical composition of the present invention that can be used as a safe and effective anti-cancer drug using various cancer cell lines.

Summary of the Invention

[0007] Summary of the Invention It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by Formula 1 or 2 described herein as an active ingredient. It is another object of the present invention to provide an N-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine derivative compound. It is an object of the present invention to provide a health functional food composition for preventing or treating cancer, comprising a compound represented by Formula 1 or 2 described herein as an active ingredient.

[0008] It is another object of the present invention to provide a method for treating cancer, the method comprising the step of administering a compound represented by Formula 1 or 2 described herein to a subject in need thereof. It is another object of the present invention to provide a compound represented by Formula 1 or 2 described herein for use in the prevention or treatment of cancer. It is another object of the present invention to provide the use of a compound represented by Formula 1 or 2 described herein for the preparation of a medicament for use in the prevention or treatment of cancer.

[0009] In order to achieve the above object, in an aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer, which contains, as an active ingredient, a compound represented by the following formula 1 or 2, its stereoisomer, its solvate, its hydrate, or its pharmaceutically acceptable salt.

[0010] [Formula 1] [Chemical formula] In the above formula 1, R is a 5- to 10-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S, a 5- to 10-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O, and S, or C 6~10 aryl;

[0011] [Formula 2] [Chemical formula] In the above formula 2, X 1 is halogen, C 1~3 alkoxy, C 1~3 alkyl, cyano, hydroxy, amino, or piperidine-substituted pyrazolyl, and X 2 is halogen or methyl-substituted piperazinyl.

[0012] In another aspect of the present invention, the present invention provides a compound represented by the following formula 1, its stereoisomer, its solvate, its hydrate, or its pharmaceutically acceptable salt.

[0013] [Formula 1] [Chemical formula] In the above formula 1, R is a 5- to 10-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O and S, a 5- to 10-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O and S, or C 6~10 is aryl.

[0014] In another aspect of the present invention, the present invention provides a health functional food composition for preventing or recovering cancer, comprising a compound represented by the above formula 1 or 2 as an active ingredient. In another aspect of the present invention, the present invention provides a method for treating cancer, said method comprising the step of administering a compound represented by the above formula 1 or 2 to a subject in need thereof.

[0015] In another aspect of the present invention, the present invention provides a compound represented by the above formula 1 or 2 for use in the prevention or treatment of cancer. In another aspect of the present invention, the present invention provides the use of a compound represented by the above formula 1 or 2 for the preparation of a medicament for use in the prevention or treatment of cancer.

[0016] Advantageous effects The compounds according to the present invention exhibit excellent anti-cancer effects against triple-negative breast cancer cell line MDA-MB-231 as well as common cancer cell lines such as A549, SK-OV-3, SK-MEL-2, HCT15, etc., and can thus be advantageously used as anti-cancer agents. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Brief description of the drawings

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[0021] Description of Preferred Embodiments The present invention is described in detail below. Embodiments of the present invention can be modified into various other forms, and the scope of the present invention is not limited to the embodiments described below. It will be fully understood by those skilled in the art having average knowledge in this field that the embodiments of the present invention are provided to more accurately interpret the present invention.

[0022] In addition, the "inclusion" of elements throughout this specification does not exclude other elements and may also include other elements unless specifically specified otherwise.

[0023] In an aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising, as an active ingredient, a compound represented by the following Formula 1 or 2, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0024] [Formula 1] [Chemical Formula] In the above Formula 1, R is a 5- to 10-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S, a 5- to 10-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O, and S, or C 6~10 aryl;

[0025] [Formula 2] [Chemical Formula] In the above formula 2, X 1 is halogen, C 1~3 alkoxy, C 1~3 alkyl, cyano, hydroxy, amino, or piperidine-substituted pyrazolyl, and X 2 is halogen, or methyl-substituted piperazinyl.

[0026] In a preferred aspect, X 1 may be bromo or piperidine-substituted pyrazolyl, and X 2 may be fluoro or methyl-substituted piperazinyl. In a further preferred aspect, the following: <1> N-(4-(Pyrrolidin-1-yl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine; <2> N-(5-Bromo-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide; and <3> 4-Fluoro-N-(5-(1-(Piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzamide There is provided a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the above formula 1 or 2 selected from the group consisting of the group of compounds, its stereoisomers, its solvates, its hydrates, or its pharmaceutically acceptable salts.

[0027] In another aspect of the present invention, the pharmaceutical composition can be used in a treatment in combination with an anti-cancer agent. The anti-cancer agent can be at least one selected from the group consisting of cisplatin, sorafenib, opdivo, tesentriq, keytruda, imfinzi, OKN-007, gefitinib, doxorubicin, vinblastine, taxol, etoposide, 5-FU, and ifosfamide. In another aspect of the present invention, the present invention provides a compound represented by the following formula 1, its stereoisomers, its solvates, its hydrates, or its pharmaceutically acceptable salts.

[0028] [Formula 1] [Chemical formula] In the above formula 1, R is a 5- to 10-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S, a 5- to 10-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O, and S, or C 6~10 aryl.

[0029] In a more preferred aspect, R may be a 5- to 6-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S, a 5- to 6-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O, and S, or phenyl; or R may be a 5-membered heterocycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S, a 5-membered heteroaryl containing one or more heteroatoms selected from the group consisting of N, O, and S, or phenyl; or R may be a 5-membered heterocycloalkyl containing one N, a 5-membered heteroaryl containing at least one N; or R may be a 5-membered heterocycloalkyl containing one N.

[0030] Most preferably, the compound represented by the above formula 1 may be a compound represented by the following formula 3. [Formula 3] [Chemical formula] The compound represented by the above formula 3 corresponds to the compound synthesized in <Example 1> described below.

[0031] In the present invention, the term "pharmaceutically acceptable salt" refers to salts commonly used in the pharmaceutical industry, such as inorganic ionic salts made from calcium, potassium, sodium, and magnesium; inorganic acid salts made from hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, and sulfuric acid; organic acid salts made from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc.; sulfonic acid salts made from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; amino acid salts made from glycine, arginine, lysine, etc.; and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but these salts do not limit the types of salts of the present invention.

[0032] In the present invention, the term "isomer" refers to compounds of the present invention or salts thereof that have the same chemical formula or molecular formula but are structurally or stereochemically different. Isomers include structural isomers such as tautomers and stereoisomers, and stereoisomers include both R isomers or S isomers (optical isomers, enantiomers) having an asymmetric carbon center and geometric isomers (trans, cis). In the present invention, all stereoisomers of the compounds represented by Formula 1 or 2 and mixtures thereof are also included within the scope of the present invention.

[0033] In the present invention, the term "hydrate" refers to the compound represented by the above Formula 1 and water bound by non-covalent intermolecular forces, and may include stoichiometric water or non-stoichiometric amounts of water. Specifically, the hydrate may contain water in a ratio of about 0.25 mol to about 10 mol based on 1 mol of the active ingredient, and more specifically, may contain about 0.5 mol, about 1 mol, about 1.5 mol, about 2 mol, about 2.5 mol, about 3 mol, about 5 mol, etc.

[0034] In the present invention, the term "solvate" refers to a compound represented by the above formula 1 and a solvent other than water bonded by non-covalent intermolecular forces, and may include stoichiometric water or non-stoichiometric amounts of water. Preferred solvents are volatile, non-harmful, and can be administered in trace amounts to humans. Specifically, the solvate may contain water in a ratio of about 0.25 moles to about 10 moles based on 1 mole of the active ingredient, and more specifically, may contain about 0.5 moles, about 1 mole, about 1.5 moles, about 2 moles, about 2.5 moles, about 3 moles, about 5 moles, and so on.

[0035] In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising, as an active ingredient, a compound represented by the above formula 1 or 2, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0036] In the present invention, the term "comprising as an active ingredient" means being included within a dosage range that brings about an effect of preventing, recovering, or treating cancer, and the dosage range may vary depending on the severity and formulation, and the number of applications may also vary depending on the age, weight, and build of the subject. In one embodiment of the present invention, in the pharmaceutical composition of the present invention, the compound represented by formula 1 is included, for example, in an amount of 0.001 mg / kg or more, preferably 0.1 mg / kg or more, more preferably 10 mg / kg or more, more preferably 100 mg / kg or more, more preferably 250 mg / kg or more, and most preferably 0.1 g / kg or more. The upper limit of the amount of the compound represented by formula 1 included in the pharmaceutical composition of the present invention may be selected within an appropriate range by those skilled in the art.

[0037] The pharmaceutical composition according to the present invention may contain, alone, an effective amount of the compound represented by formula 1 or 2, or may also include one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0038] Examples of carriers, excipients, and diluents include, but are not necessarily limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. Additionally, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may also be added and included.

[0039] The pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a subject. The pharmaceutical composition can be formulated in the form of an oral preparation, an injection preparation, or a topical preparation. Oral preparations can be selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, and lyophilized preparations, but are not necessarily limited thereto. Additionally, topical preparations can be selected from the group consisting of creams, gels, ointments, emulsions, suspensions, sprays, and transdermal patches, but are not necessarily limited thereto. The pharmaceutical composition of the present invention can be administered through various routes including oral administration, transdermal administration, subcutaneous administration, intravenous administration, or intramuscular administration.

[0040] In another aspect of the present invention, the present invention provides a health functional food composition for preventing or recovering from cancer, comprising, as an active ingredient, a compound represented by the above formula 1 or 2, its stereoisomer, its solvate, its hydrate, or its pharmaceutically acceptable salt.

[0041] In the present invention, the term "recovery" refers to the alleviation, prevention, or treatment of cancer symptoms by administering, ingesting, or applying the pharmaceutical composition or food composition of the present invention to a subject suffering from cancer. In the present invention, the term "prevention" refers to inhibiting or blocking cancer symptoms by administering, ingesting, or applying the pharmaceutical composition or food composition of the present invention to a subject who does not have cancer. In the present invention, the term "treatment" refers to complete cure of cancer symptoms, as well as partial cure, improvement, and alleviation of cancer symptoms, as a result of administering the pharmaceutical composition of the present invention to a subject suffering from cancer.

[0042] In the present invention, the term "subject" refers to any animal, including humans, that has already developed cancer or is at risk of developing cancer. The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.

[0043] In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment or improvement. The effective dosage level depends on factors including the type and severity of the subject, age, sex, drug activity, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concomitant drugs, and other factors well known in the medical field.

[0044] In the present invention, the term "administration" means providing a substance to a subject or patient by any appropriate method. The substance can be administered parenterally (for example, intravenous injection, subcutaneous injection, intraperitoneal injection, or local injection) or orally according to the desired method. The dosage range varies depending on the weight, age, sex, health status, diet, time of administration, method of administration, excretion rate, and severity of the disease of the patient. Specifically, in the present invention, the term "parenteral administration" refers to a method of administering subcutaneously, intramuscularly, intravenously, or intraperitoneally using a tube, excluding oral administration. In addition, in the present invention, the term "oral administration" refers to a method of administering an injection into the mouth to recover pathological symptoms.

[0045] For parenteral dosage forms, sterile suspensions, liquids, water-insoluble excipients, suspensions, emulsions, eye drops, eye ointments, syrups, suppositories, topical preparations such as aerosols, and sterile injection solutions can be prepared by conventional methods. Preferably, pharmaceutical compositions of creams, gels, patches, sprays, ointments, plasters, lotions, medicated dressings, eye ointments, eye drops, pastes, or poultices can be prepared, but are not necessarily limited thereto. Water-insoluble excipients and suspensions can contain, in addition to the active compound(s), propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injection esters such as ethylolate, and the like. Suppositories can contain, in addition to the active compound(s), witepsol, macrogol, tween 61, cacao butter, laurin fat, glycerogelatin, and the like.

[0046] In the present invention, the cancer can be any one selected from the group consisting of lung cancer, non-small cell lung cancer (NSCL), bronchial alveolar cell lung cancer, ovarian cancer, colorectal cancer, melanoma, gastric cancer, gastrointestinal cancer, liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, melanoma of the skin or eye, uterine cancer, rectal cancer, colon cancer, breast cancer, uterine sarcoma, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, esophageal cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, multiple myeloma, and chronic or acute leukemia. Preferably, the cancer can be liver cancer, ovarian cancer, lung cancer, colorectal cancer, melanoma, or breast cancer.

[0047] The food composition according to the present invention can be formulated in the same manner as a pharmaceutical composition and can be used as a functional food or added to various foods. Foods to which the food composition of the present invention can be added include beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice cream, and hangover relievers (drinks, low-viscosity gel types, pills, tablets, capsules, etc.), vitamin complexes, health supplements, and the like.

[0048] The food composition of the present invention contains, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, and may also include ingredients generally added during food product formulation, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the above-mentioned carbohydrates include common sugars including monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural sweeteners (thaumatin, stevia extract such as rebaudioside A, glycyrrhizin, etc.) and synthetic sweeteners (saccharin, aspartame, etc.) can be included as sweeteners. When the food composition of the present invention is formulated as a drink or beverage, in addition, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be included in addition to the compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0049] The present invention provides a health functional food or a health supplement food containing a food composition comprising, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. In the present invention, the term "health functional food or health supplement food" refers to a food manufactured and processed using raw materials or ingredients having useful functional characteristics for the human body in accordance with the Act on Health Functional Food. The term "functional" food refers to a food consumed for the purpose of obtaining useful health effects (such as regulating nutrients for the structure and function of the human body or physiological effects, etc.). In the present invention, the functional food is a food product prepared by adding a compound represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof to food materials such as beverages, tea, spices, gums, and confectioneries, or by encapsulation, powdering, or suspension. The above functional food has specific health effects when consumed, but since it is manufactured using food as a raw material, unlike general drugs, it has the advantage of not having any side effects that may occur when taking drugs continuously for a long period. The health functional food or health supplement food of the present invention thus obtained can be taken daily and is extremely useful. The amount of the compound represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof in such a health functional food or health supplement food depends on the type of the targeted health functional food, but is added within a range that does not impair the original taste of the food, and is generally in the range of 0.01 to 50 wt%, preferably 0.1 to 20 wt% of the targeted food, so it cannot be formulated uniformly. In addition, in the case of a health functional food or a health supplement food in the form of pills, granules, tablets, or capsules, it is generally added in the range of 0.1 to 100 wt%, preferably 0.5 to 80 wt%. In one aspect of the present invention, the health functional food or health supplement food of the present invention may be in the form of pills, tablets, capsules, or beverages.

[0050] The food composition of the present invention may contain conventional food additives. The suitability as a "food additive" is determined by the standards and criteria applicable to the item in question, in accordance with the general rules and general test methods for food additives approved by the Ministry of Food and Drug Safety, unless otherwise specified.

[0051] Items listed in the "Korean Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon color, licorice extract, crystalline cellulose, sorghum color, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents for noodles, preservative preparations, and tar dye preparations.

[0052] In addition, the food composition of the present invention can be prepared and processed into forms such as tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing and / or recovering from cancer.

[0053] For example, a health functional food in tablet form can be prepared by granulating in a conventional manner a mixture of a food composition containing, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, an excipient, a binder, a disintegrant, and other additives, and then compression molding the mixture with a lubricant or the like or directly compression molding the mixture. In addition, a health functional food in tablet form may optionally contain a flavors enhancer or the like and may optionally be coated with a suitable coating agent.

[0054] Among health - functional foods in capsule form, hard capsules can be prepared by filling a mixture of a food composition containing, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, and additives such as excipients, or granular materials thereof or coated granular materials thereof, into conventional hard capsules. Soft capsules can be prepared by filling a mixture of a food composition containing, as an active ingredient, a compound represented by Formula 1, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, and additives such as excipients, into a capsule base such as gelatin. Soft capsules may contain, if necessary, plasticizers such as glycerin or sorbitol, coloring agents, preservatives, and the like.

[0055] Health - functional foods in pill form can be prepared by molding, in any suitable manner, a mixture of a food composition containing, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, and excipients, binders, disintegrants, etc. If necessary, they can be coated with white sugar or another suitable coating agent, or can be coated with starch, talc, or other suitable substances.

[0056] Health - functional foods in granular form can be prepared by using, in any suitable manner, a mixture of a food composition containing, as an active ingredient, a compound represented by Formula 1 or 2, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, and excipients, binders, disintegrants, etc. If necessary, they may contain flavoring agents, flavor enhancers, and the like. When a particle - size test was conducted using No.12 (1680μm), No.14 (1410μm), and No.45 (350μm) sieves, the entire amount of the health - functional food in granular form passed through the No.12 sieve, less than 5.0% of the total amount remained on the No.14 sieve, and less than 15.0% of the total amount passed through the No.45 sieve.

[0057] There is no specific limitation on the type of food, which includes all health - functional foods in the conventional sense. Matters referred to in the pharmaceutical and food compositions of the present invention are applied in the same manner, provided they are not mutually inconsistent.

[0058] In another aspect of the present invention, the present invention provides a method for preventing or treating cancer, comprising the step of administering to a subject in need thereof a compound represented by formula 1 or 2 described herein, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0059] In another aspect of the present invention, the present invention provides a method for preventing or treating cancer, comprising the step of co-administering to a subject in need thereof a compound represented by formula 1 or 2 described herein, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, and radiation or an anticancer agent.

[0060] In another aspect of the present invention, the present invention provides the use of a compound represented by formula 1 or 2 described herein, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for use in the prevention or treatment of cancer.

[0061] In another aspect of the present invention, the present invention provides the use of a compound represented by formula 1 or 2 described herein, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, and a radiation or anticancer agent to be administered in combination, for the preparation of a medicament for use in the prevention or treatment of cancer. The detailed description of the pharmaceutical composition described above may be applied to the above methods or uses.

[0062] The compounds according to the present invention exhibit excellent anticancer effects against triple-negative breast cancer cell line MDA-MB-231 and common cancer cell lines such as MDA-MB-231 A549, SK-OV-3, SK-MEL-2, HCT15, etc., and can thus be advantageously used as anticancer agents, which is supported by the following examples and experimental examples.

[0063] In the following, the present invention will be described in detail by the following examples and experimental examples. However, the following examples and experimental examples are only for explaining the present invention, and the content of the present invention is not limited thereto.

[0064] Sample Preparation [Table A]

[0065] Example 1: N-(4-(Pyrrolidin-1-yl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [Reaction Formula 1] [Chemical Formula]

[0066] DIPEA (9.78 mL, 56.0 mmol, N,N - diisopropylethylamine) was added to a solution of 4 - chloro - 7H - pyrrolo[2,3 - d]pyrimidine (4.30 g, 28.0 mmol) and 4 - pyrrolidin - 1 - ylaniline (5.00 g, 30.8 mmol) dissolved in 1 - butanol (86 mL). Under a nitrogen atmosphere, the mixed solution was stirred at 115 °C for 24 hours. The suspension was cooled to room temperature, stirred for 1 hour, filtered, and washed with 1 - butanol (10 mL). The resulting solid was treated with EtOAc (100 mL) and water (100 mL) and stirred for 18 hours. The suspension was filtered, washed with water (10 mL) and EtOAc (10 mL), and dried under reduced pressure to give the title compound (4.41 g, 56%) as a brown solid. The MS spectrum of the compound prepared in Example 1 is shown in Figure 1, and the 1H - NMR spectrum is shown in Figure 2. As shown in Figures 1 and 2, it was confirmed that the compound of Example 1 was synthesized sufficiently.

[0067] The NMR spectra were recorded at room temperature on a Bruker BioSpin 400 MHz (AVANCE III HD 400). When collecting samples, each sample was dissolved in DMSO-d6 and transferred to an NMR tube (5-mm o.d., Wilmad, WG-1007-7).

[0068] LC-MS was performed on an Ion-trap Mass Spectrometer on an Accela-Thermo Finnigan LCQ Fleet operating in APCI(+) ionization mode.

[0069] The Agilent 1100 series was used for HPLC, and the conditions were as follows.

Table B

[0070] Example 2: N-(5-Bromo-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)benzamide [Formula 4]

Chemical formula

[0071] The compound of Example 2 was prepared with reference to Korean Patent No. 10-2129114.

[0072] Example 3: 4-Fluoro-N-(5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzamide [Formula 5]

Chemical formula

[0073] The compound of Example 3 was prepared with reference to Korean Patent No. 10-2129114.

[0074] Example 3: Evaluation of cytotoxicity against various cancer cell lines The cytotoxicity of the compound of Example 1 against various cancer cell lines was evaluated. Etoposide is sold as an anticancer drug and was used as a control drug. Specifically, the following cell viability assay was performed.

[0075] The A549, SK-OV-3, SK-MEL-2, and HCT15 cell lines were seeded in a 96-well plate at a density of 5×10 3 cells per well and cultured for 24 hours. Subsequently, the plates were treated with the compound of Example 1 or etoposide at different concentrations, followed by culturing for 24 and 48 hours. At the end of the culture, 100 μl of WST-8 solution was added to each well of the plate, and the absorbance was measured using a microplate reader with a 450 nm optical filter to measure the formazan produced by the reducing factors in the living cells. - A549: Human non-small cell lung cancer cells - SK-OV-3: Human ovarian cancer cells - SK-MEL-2: Human melanoma cells - HCT15: Human colorectal adenocarcinoma cells

[0076] The results are shown in Figure 3. Figure 3 is a series of graphs showing the results of confirming the toxicity of the compound of Example 1 of the present invention against various cancer cell lines. As shown in Figure 3, the compound of Example 1 according to the present invention exhibits excellent toxicity dose-dependently against the cancer cell lines A549, SK-OV-3, SK-MEL-2, and HCT15, and thus it can be effectively used as an active ingredient in anticancer agents.

[0077] From the above results, the compound of Example 1 according to the present invention exhibits excellent toxicity against various cancer cell lines and is confirmed to be equal to or more effective than etoposide, a topoisomerase inhibitor widely used in lung cancer, ovarian cancer, colorectal cancer, melanoma, etc. Therefore, the compound of the present invention can be effectively used as an active ingredient in various anticancer agents.

[0078] Experimental Example 2: Evaluation of breast cancer growth inhibition An experiment was conducted to confirm the tumor inhibition of the compound of Example 1, which is the active ingredient of the pharmaceutical composition of the present invention, against xenograft tumor samples of breast cancer cells (MBA-MB-231), a human-derived TNBC cell line.

[0079] Specifically, using 80 female 5-week-old xenograft Balb / c nude mice, an experiment was conducted to confirm the effect of the compound of Example 1, which is the active ingredient of the pharmaceutical composition provided by the present invention, on the inhibition of breast cancer cell growth. The experimental xenograft Balb / c nude mice lack T cells, which are immune cells, and were selected as a suitable animal model for transplanting human cancer cells. Animals within ±20% of the total average body weight at the time of administration were selected, tumor growth was checked twice a week from their surface, and the tumor size was measured with calipers for 3 weeks. In addition, the compound of Example 1 was intravenously administered at a concentration of 25 mpk, and 30% PEG was used as the solvent.

[0080] The results are shown in FIGS. 4a to 4c. FIGS. 4a to 4c are graphs showing the results of evaluating the growth inhibitory activity of the compound of Example 1 against the MDA-MB-231 TNB cell line.

[0081] From FIG. 4a showing the change in body weight following the administration of the compound of Example 1, it was confirmed that the compound of Example 1 had no biological toxicity and was safe. In particular, the cachexia phenomenon commonly observed following the administration of anticancer agents was not observed.

[0082] From FIG. 4b showing the change in tumor size following the administration of the compound of Example 1, it was confirmed that in the group administered with the compound of Example 1, tumor growth was significantly suppressed compared to the group not administered with the compound of Example 1 (CTL).

[0083] From FIG. 4c showing the change in tumor weight following the administration of the compound of Example 1, it was confirmed that in the group administered with the compound of Example 1, tumor growth was significantly suppressed compared to the group not administered with the compound of Example 1 (CTL).

[0084] From the above results, as already demonstrated in <Experimental Example 1>, the compound of Example 1 according to the present invention was found to be highly active in suppressing the growth of triple-negative breast cancer as well as various common cancer cell lines.

[0085] Experimental Example 3: Comparison of the number of mitotic figures An experiment was conducted to evaluate the tumor necrosis effect of the compound of Example 1 by measuring the number of apoptotic bodies in tumors and the H&E staining of tumor tissues in BALB / c-nude mice injected with cancer cell lines. Specifically, as shown in FIGS. 4a to 4c, to measure the number of apoptotic bodies in the tumor tissues of the T1 (twice / week) and T2 (thrice / week) groups of <Experimental Example 2>, five areas where the tumor tissues were growing were randomly selected, and the number of apoptotic bodies was measured at a magnification of x400. After calculating the sum of the measured values and expressing them as individual values, statistical comparative analysis was performed using GraphPad Prism 5.

[0086] As a result of observing apoptotic bodies, it was observed that the cells shrank and divided (blebs) and that voids (hollows) surrounded the periphery. In addition, an increase in cytoplasmic eosinophilicity was observed, and the nuclei were small, condensed or fragmented, and were phagocytosed by surrounding cells. The frequency of mitotic figures in tumor tissues is related to the dividing ability of tumor cells, which can reflect the degree of malignancy of the tumor.

[0087] The results are shown in Table 1 below and FIG. 5. FIG. 5 is a graph showing the results of comparing the number of mitotic figures in tumor tissues according to the administration of the compound of Example 1. [Table 1]

Table 1

[0088] According to the number of apoptotic bodies confirmed in Table 1 and FIG. 5, the number was dose-dependently higher in the T1 and T2 groups treated with the compound of Example 1, which is the active ingredient of the pharmaceutical composition of the present invention, compared to NC (negative control). Thus, it was confirmed that the compound of Example 1 of the present invention has a tumor-suppressing effect by increasing apoptosis of cancer cells.

[0089] Experimental Example 4: Evaluation of the inhibition of breast cancer cell lines by the compounds of Example 2 and Example 3 An experiment was conducted to confirm the growth inhibitory effect of the compounds of Examples 1 and 2, which are the active ingredients of the pharmaceutical composition of the present invention, on breast cancer cell lines through a cell viability assay.

[0090] Specifically, the following cell viability assay was performed. Human-derived breast cancer cell line BT20 and TNBC cell line MDA-MB-231 were each seeded in a 96-well plate at a density of 5×10 3 cells / well and cultured for 24 hours. Then, I-bet 762 and the compounds were treated with them at various concentrations and cultured for 48 hours. At the end of the culture, 100 μl of WST-8 solution was added to each well of the plate, and the absorbance was measured using a microplate reader with a 450 nm optical filter to measure the formazan produced by the reducing factors in living cells.

[0091] As shown in FIGS. 6 to 9, as a result of analyzing the formation of tumor-like masses by culturing the human-derived breast cancer cell line BT20 and the TNBC cell line MDA-MB-231, it was confirmed that the compound of Example 2, which is the active ingredient of the pharmaceutical composition of the present invention, suppressed the cell viability in a dose-dependent manner not only in general breast cancer cell lines but also in TNBC cell lines.

[0092] FIG. 10 shows the cell viability of MDA-MB-231, MDA-MB-453, and BT-20 after 24 hours of treatment with the compound of Example 3. As shown in FIG. 10, it was confirmed that the cell viability was suppressed in a dose-dependent manner not only in general breast cancer cell lines but also in TNBC cell lines.

[0093] Experimental Example 5: Evaluation of efficacy in combination with other anticancer agents <5-1>Evaluation of efficacy in combination with cisplatin Human ovarian cancer cell lines (SKOV-3 and OVCAR-3) were treated with control (no treatment), 0.1 μM of the compound of Example 1, 10 μM of cisplatin, and 0.1 μM of the compound of Example 1 / 10 μM of cisplatin, and the cell viability of SKOV-3 and OVCAR-3 was confirmed. The results are shown in FIG. 11.

[0094] When 0.1 μM of the compound of Example 1 or 1 μM of cisplatin was administered to SKOV-3 cell line and OVCAR-3 cell line, it was confirmed that the cell viability was suppressed. In addition, when 0.1 μM of the compound of Example 1 and 10 μM of cisplatin were administered in combination, it was also confirmed that the cell viability was further inhibited (synergistic effect).

[0095] <5-2>Evaluation of efficacy in combination with sorafenib Human liver cancer cell lines (SK Hep1 and Huh-7) were treated with control (no treatment), 5 μM of the compound of Example 1, 0.1 μM of sorafenib, and 5 μM of the compound of Example 1 / 0.1 μM of sorafenib, and the cell viability of SK Hep1 and Huh-7 was confirmed. The results are shown in Figure 12.

[0096] When 5 μM of the compound of Example 1 or 0.1 μM of sorafenib was administered to SK Hep1 cell line and Huh-7 cell line, it was confirmed that the cell viability was suppressed. In addition, when 5 μM of the compound of Example 1 and 0.1 μM of sorafenib were administered in combination, it was also confirmed that the caspase-3 activity was further inhibited (synergistic effect).

Claims

1. The following formula 1: [Formula 1] 【Chemical 1】 In the above formula 1, R is a 5- to 6-membered heterocycloalkyl containing at least 1 N, A pharmaceutical composition for use in the prevention or treatment of cancer, comprising as an active ingredient a compound represented by, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition is A pharmaceutical composition according to claim 1, comprising as an active ingredient a compound represented by formula 1 which is N-(4-(pyrrolidin-1-yl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

3. The cancer is selected from the group consisting of non-small cell lung cancer (NSCL), ovarian cancer, colorectal cancer, melanoma, breast cancer, and liver cancer, and the pharmaceutical composition according to claim 1.

4. The breast cancer is triple negative breast cancer, and the pharmaceutical composition according to claim 3.

5. The pharmaceutical composition according to claim 1, which is for use in combination therapy with an anticancer agent.

6. The anticancer agent is at least one selected from the group consisting of cisplatin, sorafenib, opdivo, tesentriq, keytruda, imfinzi, OKN-007 (Oklahoma nitron-007), gefitinib, doxorubicin, vinblastine, taxol, etoposide, 5-FU (5-fluorouracil), and ifosfamide, and the pharmaceutical composition according to claim 5.

7. The following formula 1: [Formula 1] [[Chemical 2]] In the above formula 1, R is a 5- to 6-membered heterocycloalkyl containing at least 1 N, A health functional food composition for use in the prevention or recovery of cancer, comprising as an active ingredient a compound represented by, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

8. For the preparation of a medicament for use in the prevention or treatment of cancer, formula 1: [Formula 1] 【Chemical Formula 3】 In the above formula 1, R is a 5- to 6-membered heterocycloalkyl containing at least 1 N, Use of a compound represented by, a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

9. For the preparation of a medicament for use in the prevention or treatment of cancer, formula 1: [Formula 1] [Chemical Formula 4] In the above formula 1, R is a 5- to 6-membered heterocycloalkyl containing at least 1 N, Use of a compound represented by, its stereoisomers, its solvates, its hydrates, or its pharmaceutically acceptable salts, in combination with an anti-cancer agent administered therewith.

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