N4-(indolin-7-yl)-n2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivatives

Abstract

The present invention relates to N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivatives, a method for preparing same, and a pharmaceutical composition for the prevention or treatment of cancer comprising same as an active ingredient. The pyrimidine derivatives of the present invention exhibit high inhibitory activity against EGFR mutation and HER2 mutation and thus can be usefully employed in the treatment of cancers involving EGFR or HER2 mutations.

Description

N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative

[0001] The present invention relates to an N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative, a method for preparing the same, and a pharmaceutical composition for preventing or treating cancer comprising the same as an active ingredient.

[0002]

[0003] The development of cancer is related to various environmental factors including chemicals, radiation, and viruses, as well as changes in oncogenes, tumor suppressor genes, and genes involved in apoptosis and DNA repair. Recently, understanding the molecular mechanisms of cancer has made new treatments, such as targeted anticancer therapy, possible.

[0004] Targeted therapies are generally designed to exert their effects by targeting molecules unique to cancer cells. These molecular targets include genes involved in cancer cell signal transduction pathways, angiogenesis, the cell matrix, cell cycle regulators, and apoptosis. Currently, key targeted therapies used in treatment include signal transduction pathway inhibitors, including tyrosine kinase inhibitors, and angiogenesis inhibitors.

[0005] Protein tyrosine kinases have been shown to play an important role in many malignant tumors, and in particular, the epidermal growth factor receptor (EGFR), a receptor tyrosine kinase of the erbB family, is abnormally activated in many epithelial tumors, including non-small cell lung carcinoma (NSCLC), breast cancer, glioma, squamous cell carcinoma of the head and neck, colon cancer, rectal carcinoma, head and neck cancer, gastric cancer, and prostate cancer, and activation of the EGFR-tyrosine kinase is known to cause sustained cell proliferation, invasion of surrounding tissues, distant metastasis, angiogenesis, and increased cell survival.

[0006] Specifically, the EGFR is one of the ErbB tyrosine kinase receptors family (EGFR, HER-2, ErbB-3, ErbB-4), and is a transmembrane tyrosine kinase that has an intracellular domain including an extracellular ligand-binding domain and a tyrosine kinase domain. When a ligand binds to a receptor that forms a homodimer or heterodimer, intracellular tyrosine kinase is activated, and the signal stimulated by EGFR activates the phosphatidylinositol 3-kinase (PI3K) / AKT / mTOR, RAS / RAF / MAPK, JAK / STAT) signaling pathway (Nat Rev Cancer 2007;7:169-81).

[0007] EGFR, in particular, is overexpressed in more than half of non-small cell lung cancers (NSCLC), and many studies have been conducted as a therapeutic target. EGFR tyrosine kinase inhibitors (TKIs) that suppress EGFR tyrosine kinase activity have been developed, and representative drugs include gefitinib (IRESSA™), erlotinib (TARCEVA™), lapatinib (TYKERB™, TYVERB™), and osimertinib (Tagrisso™). In particular, osimertinib is a third-generation EGFR TKI that is being used clinically. However, resistance to osimertinib has been shown when the C797S mutation, in which cysteine ​​at position 797 in EGFR exon 20 is substituted with serine, additionally occurs.

[0008] Intracellular human EGFR2 (HER2), also known as HER2 / neu or ErbB2, is a tyrosine kinase receptor belonging to the human epidermal growth factor receptor (HER / EGFR / ERBB) family, which normally participates in signal transduction pathways to induce cell growth and differentiation. HER2 shares high structural similarity with the other three EGFR family members (HER1, HER3, and HER4).

[0009] However, unlike other anticancer targets, the ligand that binds to Her2 (Human epidermal growth factor 2) is not yet known, and it partners with other HER receptors that bind to the ligand to form a heterodimer, and is involved in cell cycle increase, cell proliferation control, differentiation, and survival through various signaling pathways. In the case of antibodies targeting Her2, IgG2 type antibody therapeutics have been developed and are on the market, and the main therapeutic effect is neutralizing activity due to the antibody, not antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC).

[0010] Meanwhile, as a prior study on EGFR and / or HER2 mutant activity inhibitors, US Patent Publication No. US RE48687 E1 discloses a pyridinylaminopyrimidine derivative compound, and discloses that the compound inhibits EGFR mutant activity and cancer cell proliferation.

[0011] Accordingly, while striving to develop a cancer treatment agent that inhibits EGFR mutation and HER2 mutation, the present invention has been completed by finding that the N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative according to the present invention exhibits high inhibitory activity against EGFR mutation and HER2 mutation, and thus can be usefully used for the prevention or treatment of cancer.

[0012]

[0013] One object of the present invention is to provide a novel N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative having anticancer activity.

[0014] Another object of the present invention is to provide a method for producing an N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative.

[0015] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which contains a novel N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative having anticancer activity as an active ingredient.

[0016] Another object of the present invention is to provide a health functional food for preventing or improving cancer containing an N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative as an active ingredient.

[0017]

[0018] To achieve the above purpose,

[0019] According to one aspect of the present invention, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof is provided.

[0020] [Chemical Formula 1]

[0021]

[0022] In the above chemical formula 1,

[0023] A 1 Silver C 1-6 Alkylcarbonyl or C 1-6 alkylsulfonyl;

[0024] A 2 C is unsubstituted or substituted with one or more halogens. 1-6 alkyl;

[0025] A 3 is halogen;

[0026] R 1 is -NR b1 R b2 And,

[0027] R b1 Silver C 1-6 alkyl;

[0028] R b2is DC 1-6 C substituted with alkylamino 1-6 alkyl;

[0029] R 2 is acrylamido.

[0030] In addition, the present invention, as shown in the following reaction scheme 1,

[0031] A method for producing a compound represented by Chemical Formula 1 is provided, comprising the step of producing a compound represented by Chemical Formula 1 by reacting a compound represented by Chemical Formula 2 with a compound represented by Chemical Formula 3:

[0032] [Reaction Formula 1]

[0033]

[0034] (A in the above reaction scheme 1 1 , A 2 , A 3 , R 1 and R 2 is as defined in Chemical Formula 1 of Article 1.)

[0035] According to another aspect of the present invention, a pharmaceutical composition for preventing or treating cancer is provided, which contains a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0036] According to another aspect of the present invention, a health functional food for preventing or improving cancer is provided, which contains a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0037]

[0038] The N4-(indolin-7-yl)-N2-(2-alkoxypyridin-3-yl)pyrimidine-2,4-diamine derivative of the present invention exhibits high inhibitory activity against EGFR mutations and HER2 mutations, and thus can be usefully used in the treatment of cancer in which EGFR mutations and HER2 mutations have occurred.

[0039]

[0040] Hereinafter, the present invention will be described in detail.

[0041] One aspect of the present invention is:

[0042] A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof is provided.

[0043] [Chemical Formula 1]

[0044]

[0045] In the above chemical formula 1,

[0046] A 1 Silver C 1-6 Alkylcarbonyl or C 1-6 It is alkylsulfonyl;

[0047] A 2 C is unsubstituted or substituted with one or more halogens. 1-6 It is alkyl;

[0048] A 3 is a halogen;

[0049] R 1 is -NR b1 R b2 And,

[0050] R b1 Silver C 1-6 It is alkyl;

[0051] R b2 is DC 1-6 C substituted with alkylamino 1-6 It is alkyl;

[0052] R 2 is acrylamido.

[0053] A above 2 C is unsubstituted or substituted with one or more F 1-6 It may be alkyl, and the above A 3 can be Cl.

[0054] In another embodiment of the present invention,

[0055] A 1 is methylcarbonyl or methylsulfonyl;

[0056] A 2is CF3CH2, CH3CH2, CF3CH2CH2 or CH3;

[0057] A 3 is Cl;

[0058] R 1 is -NR b1 R b2 And,

[0059] R b1 Silver methyl;

[0060] R b2 is 2-(dimethylamino)ethyl;

[0061] R 2 is acrylamido.

[0062] Examples of the compound represented by the chemical formula 1 according to the present invention include the following compounds.

[0063] <1> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide;

[0064] <2> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide;

[0065] <3> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide;

[0066] <4> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide;

[0067] <5> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide;

[0068] <6> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide;

[0069] <7> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide;

[0070] <8> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide.

[0071] In the present invention, alkyl means a saturated hydrocarbon, and includes all straight-chain or branched-chain saturated hydrocarbons unless specifically limited. Branched-chain saturated hydrocarbons refer to saturated hydrocarbons having three or more hydrocarbon atoms. Examples of alkyl include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, and hexyl.

[0072] The compound represented by the above chemical formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and as a salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The acid addition salt is obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc.; non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc.; organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, Includes phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, etc.

[0073] The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the derivative of chemical formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., adding an organic acid or inorganic acid, filtering and drying the resulting precipitate, or by distilling the solvent and an excess acid under reduced pressure, drying, and crystallizing in the presence of an organic solvent.

[0074] Additionally, pharmaceutically acceptable metal salts can be prepared using bases. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. Among the metal salts, sodium, potassium, or calcium salts are pharmaceutically suitable. Furthermore, the corresponding salts can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable anion salt (e.g., silver nitrate).

[0075] Another aspect of the present invention is, as shown in the following reaction scheme 1,

[0076] A method for producing a compound represented by chemical formula 1 of claim 1 is provided, which comprises a step of producing a compound represented by chemical formula 1 by reacting a compound represented by chemical formula 2 with a compound represented by chemical formula 3.

[0077] [Reaction Formula 1]

[0078]

[0079] In the above reaction formula 1, A, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 is as defined in chemical formula 1 above.

[0080] In the above reaction formula, the reaction of Chemical Formula 2 and Chemical Formula 3 is a nucleophilic substitution reaction, and the reaction conditions are carried out under normal organic solvent conditions, and include organic solvents such as toluene, ethanol, dichloromethane, trifluoroacetic acid, acetonitrile, methanol, n-BuOH, tetrahydrofuran, DMF, DMSO, acetone, and chloroform, and the reaction temperature is not particularly limited, but may be carried out in the range of 0 to 100°C, 10 to 50°C, and may be carried out at room temperature.

[0081] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0082] At this time, the cancer is pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, At least one selected from the group consisting of heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid tumor, vaginal cancer, spinal cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, and thymic cancer, wherein the cancer is EGFR, It may be a cancer in which a mutation is expressed for one or more selected from the group consisting of HER2, ALK, FAK, FLT3, JAK3, KIT, and PLK4, and in one embodiment, a cancer in which a mutation is expressed for EGFR or HER2.

[0083] In addition, the compound or a pharmaceutically acceptable salt thereof can inhibit EGFR (epidermal growth factor receptor) mutations, and through such inhibition, has an activity of preventing, improving, or treating cancer. At this time, the EGFR mutations may include EGFR del19, EGFR L858R, EGFR A763_Y764insFHEA, EGFR V769_D770insASV, and EGFR D770_N771insSVD. At this time, the cancer is the same as described above.

[0084] In addition, the compound, its isomer, its solvate, its hydrate or its pharmaceutically acceptable salt can inhibit HER2 mutation and, through such inhibition, have an activity for preventing, improving or treating cancer. In this case, the HER2 mutation can include HER2 A775_G776insYVMA, etc. In this case, the cancer is the same as described above.

[0085] The compound or a pharmaceutically acceptable salt thereof has growth inhibitory activity against mutant cancer cells, which are cells expressing EGFR mutations and / or HER2 mutations as described above, and in one embodiment, may be cancer cells expressing one of EGFR del19, EGFR L858R, EGFR V769_D770insASV, EGFR D770_N771insSVD, and HER2 A775_G776insYVMA.

[0086] In addition, a pharmaceutical composition for preventing or treating cancer containing the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient can be administered as an individual therapeutic agent or used in combination with other anticancer agents in use.

[0087] In addition, a pharmaceutical composition for preventing or treating cancer containing the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient can enhance the anticancer effect by co-administering it with an anticancer agent.

[0088] The compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof can be administered in various oral and parenteral dosage forms during clinical administration. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, and emulsions. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0089] A pharmaceutical composition containing the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient can be administered parenterally, and parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

[0090] At this time, in order to formulate a dosage form for parenteral administration, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof is mixed with a stabilizer or buffer in water to prepare a solution or suspension, which can be prepared in an ampoule or vial unit dosage form. The composition may be sterilized and / or contain auxiliary agents such as preservatives, stabilizers, wetting agents or emulsifying promoters, salts for osmotic pressure control and / or buffers, and other therapeutically useful substances, and may be formulated according to conventional mixing, granulation or coating methods.

[0091] Oral dosage forms include, for example, tablets, pills, hard / soft capsules, solutions, suspensions, emulsions, syrups, granules, elixirs, and troches, which contain, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine), lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). Tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidine, and, if desired, disintegrating agents or effervescent mixtures such as starch, agar, alginic acid or its sodium salt, and / or absorbents, coloring agents, flavoring agents, and sweetening agents.

[0092] Another aspect of the present invention provides a health functional food for preventing or improving cancer, comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. In this case, the cancer is the same as described above.

[0093] The compound represented by the above chemical formula 1 according to the present invention can be added directly to food or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient mixed can be appropriately determined depending on its purpose of use (prevention or improvement). Generally, the amount of the compound in a health food can be added in an amount of 0.1 to 90 parts by weight based on the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can also be used in an amount exceeding the above range.

[0094] Another aspect of the present invention provides a method for preventing, improving or treating cancer, comprising administering to a subject in need thereof a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0095] Another aspect of the present invention provides a method for preventing, improving or treating cancer, comprising administering to a subject in need thereof a pharmaceutical composition or health functional food containing a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0096] Another aspect of the present invention provides the use or use of a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof in the prevention, improvement or treatment of cancer.

[0097] The compound represented by the above chemical formula 1 of the present invention or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered in a “pharmaceutically effective amount.” 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, and the effective dosage level can be determined according to factors including the subject type and severity, age, sex, activity of the drug, sensitivity to the drug, administration time, administration route and excretion rate, treatment period, concurrently used drugs, and other factors well known in the medical field. For example, effective amounts of 0.001 mg / kg to 1000 mg / kg, 0.01 mg / kg to 100 mg / kg, or 0.1 to 20 mg / kg, or 0.1 to 500 mg / kg are included. The amount of the pharmaceutical composition of the present invention can be selected and implemented within an appropriate range by a person skilled in the art.

[0098]

[0099] Hereinafter, the present invention will be described in detail through examples and experimental examples.

[0100] However, the following examples and experimental examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples and experimental examples.

[0101] The following discloses a manufacturing example for the preparation of the example compound.

[0102]

[0103] <Manufacturing Examples 1 to 4> were manufactured according to the scheme below.

[0104]

[0105] <Manufacturing Example 1> Preparation of 6-chloro-3-nitro-2-(2,2,2-trifluoroethoxy)pyridine

[0106] To a stirred solution of 2,6-dichloro-3-nitropyridine (5.0 g, 25.9 mmol, 1.0 eq) in toluene (50 mL) were added sodium hydride (1.3 g, 31.0 mmol, 1.2 eq) and trifluoroethanol (2.0 mL, 28.5 mmol, 1.1 eq) at 0 °C, and the resulting mixture was stirred at room temperature for 12 h. The mixture was quenched by the addition of water, extracted with ethyl acetate, and the organic layer was washed with water and dried over Na2SO4. The obtained crude material was concentrated under reduced pressure and the desired Preparation Example 1 was obtained (2.1 g, quantitative) without further purification.

[0107] 1 H NMR (300 MHz, CDCl3) δ 6.95 (d,J= 7.9 Hz, 1H), 6.85 (d,J= 7.9 Hz, 1H), 4.79 (q,J= 8.4 Hz, 2H), 3.85 (s, 2H).

[0108]

[0109] <Manufacturing Example 2> Preparation of 6-chloro-2-(2,2,2-trifluoroethoxy)pyridin-3-amine

[0110] To a stirred solution of Preparation Example 1 (5.8 g, 22.8 mmol, 1.0 eq) in ethanol (40 mL) and water (15 mL) were added ammonium chloride (6.0 g, 114 mmol, 5.0 eq) and iron powder (6.4 g, 114 mmol, 5.0 eq) at room temperature. The resulting mixture was heated to 80°C overnight. The reaction solution was filtered through Celite and concentrated to remove ethanol, and the residue was extracted with ethyl acetate. The organic layer was dried over sodium sulfate, concentrated, and purified by column chromatography using Hex / EA (1:1) as an eluent to give the desired product Preparation Example 2 as a tan solid (3.4 g, 67%).

[0111] 1H NMR (300 MHz, CDCl3) δ 6.95 (d,J= 7.9 Hz, 1H), 6.85 (d,J= 7.9 Hz, 1H), 4.79 (q,J= 8.4 Hz, 2H), 3.85 (s, 2H).

[0112]

[0113] <Manufacturing Example 3> Preparation of N-(6-chloro-2-(2,2,2-trifluoroethoxy)pyridin-3-yl)acetamide

[0114] Acetyl chloride (1.6 mL, 22.5 mmol, 1.5 eq) was slowly added to a stirred solution of Preparation Example 2 (3.4 g, 15.0 mmol, 1.0 eq) and diisopropylethylamine (3.9 mL, 22.5 mmol, 1.5 eq) in dichloromethane (30 mL) at 0°C. The resulting mixture was stirred for 3 hours. The reaction mixture was washed sequentially with water, sodium bicarbonate, and saturated sodium chloride solution, dried over sodium sulfate, and concentrated to obtain a crude product, which was purified by silica gel column chromatography using Hex / EA (6:4) as an eluent to obtain the desired product Preparation Example 3 as a yellow-brown solid (2.6 g, 65%).

[0115] 1 H NMR (300 MHz, CDCl3) δ 8.69 (d,J= 8.3 Hz, 1H), 7.05 (d,J= 8.3 Hz, 1H), 4.84 (q,J= 8.3 Hz, 2H), 2.27 (s, 3H).

[0116]

[0117] <Manufacturing Example 4> Preparation of N-(6-chloro-5-nitro-2-(2,2,2-trifluoroethoxy)pyridin-3-yl)acetamide

[0118] Fuming nitric acid (0.31 mL, 10.6 mmol, 1.1 eq) was slowly added dropwise to a stirred solution of Preparation Example 3 (2.6 g, 9.6 mmol, 1.0 eq) dissolved in trifluoroacetic anhydride (15 mL) at -10°C. The reaction mixture was stirred overnight. The reaction mixture was slowly added to crushed ice, and the precipitated solid was filtered. The obtained product was dried at 60°C and triturated with ethyl acetate to give the desired product Preparation Example 4 as a yellowish-brown solid (1.8 g, 60%).

[0119] 1 H NMR (300 MHz, CDCl3) δ 9.41 (s, 1H), 4.95 (q,J= 8.1 Hz, 2H), 2.32 (s, 3H).

[0120]

[0121] <Manufacturing Examples 5 to 8> were manufactured according to the scheme below.

[0122]

[0123] <Manufacturing Example 5> Preparation of N-(6-((2-(dimethylamino)ethyl)(methyl)amino)-5-nitro-2-(2,2,2-trifluoroethoxy)pyridin-3-yl)acetamide

[0124] To a stirred solution of Preparation Example 4 (870 mg, 2.7 mmol, 1.0 eq) dissolved in acetonitrile (10 mL) was added N 1 ,N 1 ,N 2 -Trimethylethane-1,2-diamine (633 μL, 4.89 μmol, 1.2 eq) was added and heated at 80°C for 3 hours. The reaction solution was concentrated under reduced pressure to approximately 1 / 3 of the volume, and 5.0 mL of ethyl acetate was added to precipitate a solid. The solid was filtered and dried to obtain the desired product, Preparation Example 5, as a yellow solid (750 mg, 71%).

[0125] 1H NMR (300 MHz, DMSO) δ 9.54 (s, 1H), 8.61 (s, 1H), 5.16 (q,J= 9.0 Hz, 2H), 4.01 (s, 2H), 2.83 (d,J= 5.8 Hz, 9H), 2.07 (s, 3H).

[0126] <Manufacturing Example 6> Preparation of N-(5-amino-6-((2-(dimethylamino)ethyl)(methyl)amino)-2-(2,2,2-trifluoroethoxy)pyridin-3-yl)acetamide

[0127] To a stirred solution of Preparation Example 5 (670 mg, 4.0 mmol) dissolved in methanol (15 mL) was added 10% Pd / C (67 mg) under an argon atmosphere. The resulting mixture was stirred at room temperature for 14 hours under 1 atm hydrogen gas pressure. The reaction mixture was filtered through Celite and concentrated under reduced pressure to yield the desired Preparation Example 6 as a reddish brown oil (670 mg, quantitative).

[0128] 1 H NMR (300 MHz, CDCl3) δ 8.11 (s, 1H), 7.34 (s, 1H), 4.74 (q,J= 8.6 Hz, 2H), 3.51 (t,J= 5.8 Hz, 2H), 3.11 (t,J= 5.9 Hz, 2H), 2.79 (d,J= 15.8 Hz, 9H), 2.22 (s, 3H).

[0129] <Manufacturing Example 7> Preparation of N-(5-acetamido-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide

[0130] Acryloyl chloride (208 mg, 2.3 mmol, 1.2 eq) was added to a stirred solution of Preparation Example 6 (670 mg, 1.9 mmol, 1.0 eq) and triethylamine (0.4 mL, 2.8 mmol, 1.5 eq) in DCM (10 mL) at 0°C. The reaction mixture was slowly warmed to room temperature and maintained for 3 h. The reaction mixture was diluted with DCM and quenched with an aqueous sat. NaHCO3 solution. The organic layer was separated, washed with water and brine, dried over Na2SO4, and evaporated under reduced pressure. The obtained crude material was purified by column chromatography using DCM / MeOH (90:10) as an eluent to give the desired product Preparation Example 7 as a tan solid (300 mg, 39%).

[0131] 1 H NMR (300 MHz, DMSO) δ 10.04 (s, 1H), 9.31 (s, 1H), 8.20 (d,J= 2.7 Hz, 1H), 6.69 (s, 1H), 6.27 (dd,J= 17.0, 2.0 Hz, 1H), 5.83 - 5.67 (m, 1H), 5.00 (q,J= 9.2 Hz, 2H), 3.51 (s, 2H), 3.32 (s, 3H), 2.79 (s, 3H), 2.60 (s, 5H), 2.05 (s, 3H).

[0132] <Manufacturing Example 8> Preparation of N-(5-amino-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide

[0133] To a stirred solution of Preparation Example 7 (150 mg, 0.386 mmol) dissolved in methanol (3 mL) was added concentrated hydrochloric acid (0.15 mL) and heated at 60°C for 3 hours. After completion of the reaction, the crude material was concentrated under reduced pressure, and the desired Preparation Example 8 was obtained without further purification.

[0134] MS(ESI) calculated for C 15 H 25N5O2361.3, found 362.3 [M+H + ].

[0135] <Manufacturing Example 9> Preparation of N-(5-amino-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide

[0136]

[0137] Compound of Manufacturing Example 9 was obtained in a similar manner to Manufacturing Examples 1 to 8.

[0138] MS(ESI) calculated for C 15 H 25 N5O2307.4, found 308.4 [M+H + ].

[0139] <Manufacturing Example 10> Preparation of N-(5-amino-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide

[0140]

[0141] Compound 10 of Manufacturing Example was obtained in a similar manner to Manufacturing Examples 1 to 8.

[0142] MS(ESI) calculated for C 16 H 24 F3N5O2375.1, found 376.1 [M+H + ].

[0143] <Manufacturing Example 11> Preparation of N-(5-amino-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide

[0144]

[0145] Compound 11 of Manufacturing Example was obtained in a similar manner to Manufacturing Examples 1 to 8.

[0146] MS(ESI) calculated for C15H25N5O2 307.4, found 308.4 [M+H+].

[0147]

[0148] <Example 1> Preparation of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide

[0149]

[0150] Preparation Example 8 (41.3 mg 0.11 mmol) was added to a stirred solution of (1-(7-((2,5-dichloropyrimidin-4-yl)amino)indolin-1-yl)ethan-1-one (37.0 mg 0.11 mmol) [KR 10-2023-0022393] dissolved in 1 N TFA n-BuOH (1.0 mL) at room temperature. The resulting mixture was heated at 90°C for 16 h. The residue was diluted with CH2Cl2 and neutralized by adding saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol, and filtration gave the pure product. Example 1 was obtained as a yellowish brown solid (25 mg, 30%).

[0151] 1H NMR (300 MHz, DMSO) δ 9.81 (d,J= 17.0 Hz, 1H), 8.22 (s, 1H), 8.06 (d,J= 7.6 Hz, 2H), 7.56 (d,J= 7.5 Hz, 1H), 7.02 - 6.84 (m, 2H), 6.47 (dd,J= 17.2, 10.2 Hz, 1H), 6.22 (dd,J= 17.1, 2.1 Hz, 1H), 5.77 (dd,J= 10.0, 2.2 Hz, 1H), 4.85 (q,J= 9.1 Hz, 2H), 4.11 (t,J= 7.6 Hz, 2H), 3.32 (s, 3H), 3.18 (s, 2H), 3.03 (t,J= 7.7 Hz, 2H), 2.84 (s, 2H), 2.31 (s, 2H), 2.23 (s, 5H); MS(ESI) calculated for C 29 H 33 ClF3N9O3647.2, found 648.2 [M+H + ].

[0152] <Example 2> Preparation of N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide

[0153]

[0154] To a stirred solution of N-(2,5-dichloropyrimidin-4-yl)-1-(methylsulfonyl)indolin-7-amine (61 mg, 0.16 mmol) [KR 10-2023-0022393] in 1 N TFA dissolved in BuOH (1.2 mL) was added Preparation 8 (61 mg, 0.16 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to obtain pure Example 2 as a yellow-brown solid (30 mg, 26%).

[0155] 1 H NMR (300 MHz, DMSO) δ 9.84 (s, 1H), 8.95 (s, 1H), 8.41 (s, 1H), 8.11 (s, 1H), 8.09 - 8.02 (m, 1H), 7.74 (d,J= 7.9 Hz, 1H), 7.04 (dd,J= 17.5, 7.3 Hz, 2H), 6.47 (dd,J= 17.0, 10.1 Hz, 1H), 6.22 (dd,J= 17.0, 2.1 Hz, 1H), 5.81 - 5.71 (m, 1H), 4.86 (q,J= 9.0 Hz, 2H), 4.04 (t,J= 7.4 Hz, 2H), 3.32 (s, 3H), 3.17 (s, 2H), 3.09 (d,J= 7.4 Hz, 1H), 3.04 (s, 3H), 2.86 (d,J= 3.0 Hz, 2H), 2.21 (s, 6H); MS(ESI) calculated for C 28 H 33 ClF3N8O4S 683.2, found 684.2 [M+H + ].

[0156] <Example 3> Preparation of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide

[0157]

[0158] To a stirred solution of (1-(7-((2,5-dichloropyrimidin-4-yl)amino)indolin-1-yl)ethan-1-one (75.0 mg, 0.2 mmol) in 1 N TFA dissolved in n-BuOH (1.1 mL) was added Preparation 9 (71.3 mg, 0.2 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to give pure Example 3 as a tan solid. Obtained (28 mg, 20%).

[0159] 1H NMR (300 MHz, DMSO) δ 9.80 (d,J= 12.3 Hz, 2H), 8.15 - 8.05 (m, 2H), 7.90 (s, 1H), 7.62 (d,J= 7.8 Hz, 1H), 6.98 (d,J= 6.6 Hz, 2H), 6.51 (dd,J= 17.1, 9.7 Hz, 1H), 6.23 (d,J= 16.9 Hz, 1H), 5.78 (d,J= 8.5 Hz, 1H), 4.31 (q,J= 7.0 Hz, 2H), 4.12 (t,J= 7.7 Hz, 2H), 3.04 (t,J= 7.6 Hz, 2H), 2.78 (s, 3H), 2.31 (s, 7H), 1.28 (t,J= 7.0 Hz, 3H); MS(ESI) calculated for C 29 H 36 ClN9O3593.2, found 594.2 [M+H + ].

[0160] <Example 4> Preparation of N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide

[0161]

[0162] To a stirred solution of N-(2,5-dichloropyrimidin-4-yl)-1-(methylsulfonyl)indolin-7-amine (69.4 mg, 0.1 mmol) in 1 N TFA and n-BuOH (1.2 mL) was added Preparation 9 (59.3 mg, 0.1 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to obtain pure Example 4 as a yellowish brown solid (26 mg, 21%).

[0163] 1 H NMR (300 MHz, DMSO) δ 9.79 (s, 1H), 8.96 (s, 1H), 8.12 (d,J= 5.0 Hz, 2H), 7.79 (d,J= 7.4 Hz, 1H), 7.06 (q,J= 8.4 Hz, 2H), 6.52 (dd,J= 17.1, 10.0 Hz, 1H), 6.22 (d,J= 16.8 Hz, 1H), 5.83 - 5.71 (m, 1H), 4.31 (q,J= 6.9 Hz, 2H), 4.05 (t,J= 7.2 Hz, 2H), 3.18 (d,J= 5.1 Hz, 2H), 3.09 (d,J= 7.0 Hz, 5H), 2.80 (d,J= 3.8 Hz, 3H), 2.31 (d,J= 18.0 Hz, 5H), 1.27 (t,J= 6.9 Hz, 3H); MS(ESI) calculated for C 28 H 36 ClN9O4S 629.2, found 630.2 [M+H + ].

[0164] <Example 5> Preparation of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide

[0165]

[0166] To a stirred solution of (1-(7-((2,5-dichloropyrimidin-4-yl)amino)indolin-1-yl)ethan-1-one (80.5 mg, 0.2 mmol) in 1 N TFA dissolved in n-BuOH (1.1 mL) was added Preparation 10 (70.0 mg, 0.2 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to give pure Example 5 as a yellowish brown solid. It was obtained as a solid (30 mg, 21%).

[0167] 1H NMR (400 MHz, DMSO) δ 9.85 - 9.77 (m, 2H), 8.17 (s, 1H), 8.10 - 8.00 (m, 1H), 7.90 (s, 0H), 7.59 (d,J= 5.3 Hz, 1H), 6.98 - 6.92 (m, 2H), 6.48 (dd,J= 17.2, 10.2 Hz, 1H), 6.22 (d,J= 17.2 Hz, 1H), 5.77 (d,J= 10.1 Hz, 1H), 4.46 (t,J= 6.2 Hz, 2H), 4.11 (t,J= 7.7 Hz, 2H), 3.31 (s, 3H), 3.17 - 3.13 (m, 2H), 3.03 (t,J= 7.8 Hz, 2H), 2.80 (s, 3H), 2.80 - 2.65 (m, 3H), 2.31 (s, 3H), 2.24 (s, 6H); MS(ESI) calculated for C 30 H 35 ClF3N9O3661.2, found 662.2 [M+H + ].

[0168] <Example 6> Preparation of N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide

[0169]

[0170] To a stirred solution of N-(2,5-dichloropyrimidin-4-yl)-1-(methylsulfonyl)indolin-7-amine (89.0 mg, 0.2 mmol) in 1 N TFA and n-BuOH (1.2 mL) was added Preparation 10 (93.0 mg, 0.2 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to obtain pure Example 6 as a yellowish brown solid (35 mg, 20%).

[0171] 1 H NMR (400 MHz, DMSO) δ 9.80 (s, 1H), 8.96 (s, 1H), 8.16 - 8.08 (m, 4H), 7.76 (d,J= 8.0 Hz, 1H), 7.10 - 6.97 (m, 3H), 6.47 (dd,J= 17.0, 10.3 Hz, 1H), 6.25 - 6.16 (m, 1H), 5.76 (d,J= 10.2 Hz, 1H), 4.45 (t,J= 6.3 Hz, 3H), 4.04 (t,J= 7.4 Hz, 3H), 3.31 (s, 3H), 3.19 - 3.15 (m, 2H), 3.11 - 3.03 (m, 5H), 2.82 (s, 3H), 2.77 - 2.68 (m, 2H), 2.26 - 2.22 (m, 6H); MS(ESI) calculated for C 29 H 35 ClF3N9O4S 629.1, found 630.1 [M+H + ].

[0172] <Example 7> Preparation of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide

[0173]

[0174] To a stirred solution of (1-(7-((2,5-dichloropyrimidin-4-yl)amino)indolin-1-yl)ethan-1-one (191 mg, 0.5 mmol) in 1 N TFA dissolved in n-BuOH (2.0 mL) was added Preparation 11 (158 mg, 0.5 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to give pure Example 7 as a tan solid. Obtained (28 mg, 20%).

[0175] 1H NMR (300 MHz, DMSO) δ 9.83 - 9.73 (m, 2H), 8.14 - 8.01 (m, 3H), 7.67 - 7.58 (m, 1H), 7.03 - 6.93 (m, 2H), 6.46 (dd,J= 16.9, 10.2 Hz, 1H), 6.21 (dd,J= 17.1, 2.2 Hz, 1H), 5.75 (dd,J= 10.1, 2.2 Hz, 1H), 4.11 (t,J= 7.7 Hz, 2H), 3.80 (s, 3H), 3.46 - 3.35 (m, 2H), 3.19 - 2.90 (m, 4H), 2.81 (d,J= 7.8 Hz, 3H), 2.50 (s, 2H), 2.31 (s, 3H), 2.26 - 2.17 (m, 6H); MS(ESI) calculated for C 28 H 34 ClN9O3579.2, found 580.2 [M+H + ].

[0176] <Example 8> Preparation of N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide

[0177]

[0178] To a stirred solution of N-(2,5-dichloropyrimidin-4-yl)-1-(methylsulfonyl)indolin-7-amine (210 mg, 0.5 mmol) in 1 N TFA and n-BuOH (2.0 mL) was added Preparation 11 (156 mg, 0.5 mmol) at room temperature. The resulting mixture was heated to 90°C for 16 h. The reaction mixture was cooled and concentrated under reduced pressure. The residue was diluted with CH2Cl2 and neutralized by the addition of saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with CH2Cl2. The organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography using 1–15% methanol in DCM as the eluent. Further purification was performed by trituration with methanol and filtration to obtain pure Example 8 as a yellowish brown solid (26 mg, 21%).

[0179] 1 H NMR (300 MHz, DMSO) δ 9.77 (s, 1H), 8.96 (s, 2H), 8.25 (s, 1H), 8.15 - 8.02 (m, 2H), 7.81 (d,J= 7.8 Hz, 2H), 7.10 - 7.02 (m, 2H), 6.49 (dd,J= 16.9, 10.1 Hz, 1H), 6.22 (d,J= 17.2 Hz, 1H), 5.81 - 5.71 (m, 1H), 4.05 (t,J= 7.2 Hz, 2H), 3.83 (s, 3H), 3.15 - 2.96 (m, 6H), 2.89 - 2.74 (m, 4H), 2.40 - 2.18 (m, 6H); MS(ESI) calculated for C 27 H 34 ClN9O4S 615.2, found 616.2 [M+H + ].

[0180] The structural formulas of compounds of Examples 1 to 8 manufactured according to the present invention are summarized and shown in Table 1 below.

[0181]

[0182] <Experimental Example 1> Measurement of the inhibitory ability of the compound represented by Chemical Formula 1 according to the present invention against EGFR mutant enzyme

[0183] To confirm the inhibitory ability of the compound represented by Chemical Formula 1 according to the present invention against the EGFR mutant enzyme, the following experiment was performed. The results are shown in Table 2 below.

[0184] The activity measurement for the EGFR mutant enzyme of the compound of the present invention was performed as follows using the HTRF system sold by Cisbio. As the EGFR mutant enzyme, the EGFR del19 enzyme was purchased as a recombinant protein provided by Carna Biosciences, and the EGFR A763_Y764insFHEA enzyme was purchased as a recombinant protein provided by SignalChem.

[0185] The composition of the assay buffer used for activity measurement was 50 mM Tris-HCl pH 7.5, 100 mM NaCl, 7.5 mM MgCl2, 3 mM KCl, 0.01% Tween 20, 0.1% BSA, and 1 mM DTT. The enzyme reaction was performed using 50 mM ATP and 0.5 mM biotin-labeled peptide substrate. The analysis of the compound's inhibitory effect on EGFR activity was performed according to the following assay reaction recipe.

[0186] Component 1: 4 uL of EGFR mutant enzyme

[0187] Component 2: 2 uL of compound solution

[0188] Component 3: 4 uL ATP and biotin-labeled peptide

[0189] The enzyme reaction is started by mixing component 1 and component 2 first and then adding component 3. After 2 hours of reaction at 37°C, 10 mL of a measuring solution consisting of streptavidin-XL665 and europium-labeled anti-phosphotyrosine antibody provided by Cisbio is added to the enzyme reaction solution and reacted at room temperature for 1 hour. Finally, the enzyme activity is quantitatively measured by obtaining the ratio of the fluorescence values ​​at 615 nm and 665 nm using an Envision device from Perkin-Elmer and the inhibitory activity of the compound is confirmed. The measured values ​​at seven compound concentrations are analyzed using the Prism program (version 5.01, Graphpad Software, Inc.) and the inhibitory activity index of the compound, IC 50 (uM) value was calculated.

[0190] Example EGFR del19 IC 50 (uM)EGFR A763_Y764insFHEA IC 50 (uM)Osimertinib<0.0010.0071<0.0010.0022<0.001<0.0013<0.0010.01 24<0.0010.0025<0.0010.0046<0.001<0.0017<0.0010.0048<0.001<0.001

[0191] As shown in Table 2, it was confirmed that the example compound according to the present invention exhibited high inhibitory ability against various EGFR mutations including EGFR del19 and EGFR A763_Y764insFHEA mutations.

[0192]

[0193] <Experimental Example 2> Measurement of the inhibitory ability of the compound represented by Chemical Formula 1 according to the present invention against HER2 mutation

[0194] To confirm the inhibitory ability of the compound represented by Chemical Formula 1 according to the present invention against HER2 mutations, the following experiments were performed. The results are shown in Table 3 below.

[0195] The activity measurement for the HER mutant enzyme of the compound of the present invention was performed as follows using the HTRF system sold by Cisbio. As the HER2 mutant enzyme, the HER2 A775_G776insYVMA mutant enzyme was purchased as a recombinant protein provided by Carna Biosciences.

[0196] The composition of the assay buffer used for activity measurement was 50 mM Tris-HCl pH 7.5, 100 mM NaCl, 7.5 mM MgCl2, 3 mM KCl, 0.01% Tween 20, 0.1% BSA, and 1 mM DTT. The enzyme reaction was performed using 50 mM ATP and 0.5 mM biotin-labeled peptide substrate. The activity inhibitory effect of the compound on the HER2 A775_G776insYVMA mutant was analyzed according to the following assay reaction recipe.

[0197] Component 1: 4 uL of HER2 A775_G776insYVMA mutant enzyme

[0198] Component 2: 2 uL of compound solution

[0199] Component 3: 4 uL ATP and biotin-labeled peptide

[0200] The enzyme reaction is started by mixing component 1 and component 2 first and then adding component 3. After 2 hours of reaction at 37°C, 10 mL of a measuring solution consisting of streptavidin-XL665 and europium-labeled anti-phosphotyrosine antibody provided by Cisbio is added to the enzyme reaction solution and reacted at room temperature for 1 hour. Finally, the enzyme activity is quantitatively measured by obtaining the ratio of the fluorescence values ​​at 615 nm and 665 nm using an Envision device from Perkin-Elmer and the inhibitory activity of the compound is confirmed. The measured values ​​at seven compound concentrations are analyzed using the Prism program (version 5.01, Graphpad Software, Inc.) and the inhibitory activity index of the compound, IC 50 (uM) value was calculated.

[0201] Example HER2 A775_G776insYVMA IC 50 (uM)Osimertinib1.01<0.0012<0.0013<0.0014<0.00150.01160.00370.01580.005

[0202] As shown in Table 3, it was confirmed that the example compound according to the present invention exhibited high inhibitory ability against various HER2 mutations including the HER2 A775_G776insYVMA mutation.

[0203]

[0204] <Experimental Example 3> Measurement of the EGFR mutant cell growth inhibition ability of the compound represented by chemical formula 1 according to the present invention

[0205] To confirm the inhibitory ability of the compound represented by Chemical Formula 1 according to the present invention against EGFR mutant cell growth, the following experiment was performed. The results are shown in Table 4 below.

[0206] The activity of the compounds of the present invention against PC9 EGFR del19 and YU-1196 EGFR D770_N771insSVD mutant cell lines was measured using the CellTiter-Glo system sold by Promega as follows. The CellTiter-Glo assay is a method for confirming cell viability by measuring ATP present in cells in a cell culture state. PC9 cells were purchased from Cellosaurus and used, and YU-1163 (D770_N771insSVD) cells were established using patient-derived cells at Yonsei Medical Center in Sinchon, Seoul. All mutant cell lines were cultured in RPMI containing 10% FBS and 1% penicillin-streptomycin with 1 ug / ml of puromycine at 37℃ in a 5% CO2 incubator.

[0207] The analysis of the compound's inhibitory effect on EGFR mutant cell growth was conducted according to the following assay reaction recipe. 2000 or 2500 cells / 100 μL were subcultured in a 96-well cell culture plate, and after 24 hours, the compound represented by Chemical Formula 1 was treated at concentrations of 0, 0.1, 0.3, 1, 3, 10, 30, 100, 300, and 1000 (nM). After 72 hours of reaction, the compound-treated plate was incubated at room temperature for 30 minutes, then 100 μL of the reagent was additionally treated and shaken at room temperature for 10 minutes. Finally, the ratio of the fluorescence value at 570 nm was obtained using equipment to quantitatively measure and confirm the compound's inhibitory effect on cell growth. The measured values ​​at 10 compound concentrations were analyzed using the Prism program (version 5.01, Graphpad Software, Inc.) and IC, an indicator of the cell growth inhibition ability of the compound, was calculated. 50 (nM) values ​​were calculated.

[0208] Example PC9 EGFR del19 IC 50(nM)YU-1163 EGFR D770_N771insSVD IC 50 (nM)Osimertinib6.320011.82421.51632.02641.92352.66162.22672.42781.923

[0209] As shown in Table 4, it was confirmed that the example compound according to the present invention exhibited high inhibitory ability against cell growth of various EGFR mutants including PC9 EGFR del19 and YU-1163 EGFR D770_N771insSVD mutants.

[0210]

[0211] <Experimental Example 4> Measurement of the inhibitory ability of the compound represented by chemical formula 1 according to the present invention against HER2 mutation in Ba / F3 cell line

[0212] In order to confirm the inhibitory effect of the compound represented by chemical formula 1 according to the present invention on HER2 mutant cell growth in the Ba / F3 cell line, the following experiment was performed. The results are shown in Table 5 below.

[0213] The activity of the compound of the present invention against the Ba / F3 HER2 A775_G776insYVMA mutant cell line was measured using the CellTiter-Glo system sold by Promega as follows. The CellTiter-Glo assay is a method for confirming cell viability by measuring ATP present in cells in a cell culture state. The Ba / F3 HER2 A775_G776insYVMA mutant cell line was purchased and used as a cell line provided by Signosis. The Ba / F3 HER2 A775_G776insYVMA mutant cell line was cultured in a CO2 incubator at 37°C with 1 ug / ml of puromycine added to RPMI containing 10% FBS and 1% penicillin-streptomycin.

[0214] The analysis of the compound's inhibitory effect on HER2 mutant cell growth was conducted according to the following assay reaction recipe. 2000 or 2500 cells / 100 μL were passaged in a 96-well cell culture plate and cultured, and after 24 hours, the compound represented by Chemical Formula 1 was treated at concentrations of 0, 0.1, 0.3, 1, 3, 10, 30, 100, 300, and 1000 (nM). After 72 hours of reaction, the compound-treated plate was incubated at room temperature for 30 minutes, then 100 μL of the reagent was additionally treated and shaken at room temperature for 10 minutes. Finally, the ratio of the fluorescence value at 570 nm was obtained using equipment to quantitatively measure and confirm the compound's inhibitory effect on cell growth. The measured values ​​at 10 compound concentrations were analyzed using the Prism program (version 5.01, Graphpad Software, Inc.) and IC, an indicator of the cell growth inhibition ability of the compound, was calculated. 50 (nM) values ​​were calculated.

[0215] Example Ba / F3 HER2 A775_G776insYVMA IC 50 (nM)Osimertinib29011128.6315416522622719816

[0216] As shown in Table 5, it was confirmed that the example compound according to the present invention exhibits a high inhibitory ability against the cell growth of various HER2 mutations, including the Ba / F3 HER2 A775_G776insYVMA mutation, which is a HER2 mutation. Therefore, it was confirmed that the compound represented by Chemical Formula 1 according to the present invention not only exhibits a high inhibitory ability against EGFR mutations, but also has a high growth inhibitory ability against cells expressing EGFR mutations, and from this, it can be usefully used in the treatment of cancer expressing EGFR mutations such as EGFR del19 and EGFR A763_Y764insFHEA, PC9 EGFR del19, and YU-1196 EGFR D770_N771insSVD.

[0217] In addition, it was confirmed that the compound represented by chemical formula 1 according to the present invention not only exhibits high inhibitory ability against HER2 mutations, but also has high growth inhibitory ability against cells expressing HER2 mutations, and thus can be usefully used in the treatment of cancer expressing HER2 mutations such as HER2 A775_G776insYVMA.

[0218] Through the above results, it can be seen that the pyrimidine derivative compound according to the present invention can effectively suppress EGFR and HER2 mutations, and thus can be usefully used as a pharmaceutical composition for preventing or treating cancer.

Claims

1. A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, A 1 Silver C 1-6 Alkylcarbonyl or C 1-6 alkylsulfonyl; A 2 C is unsubstituted or substituted with one or more halogens. 1-6 alkyl; A 3 is halogen; R 1 is -NR b1 R b2 And, R b1 Silver C 1-6 alkyl; R b2 is DC 1-6 C substituted with alkylamino 1-6 alkyl; R 2 is acrylamido.

2. In paragraph 1, A 1 Silver C 1-3 Alkylcarbonyl or C 1-3 alkylsulfonyl; A 2 C is unsubstituted or substituted with one or more halogens. 1-3 alkyl; A 3 is halogen; R 1 is -NR b1 R b2 And, R b1 Silver C 1-3 alkyl; R b2 is DC 1-3 C substituted with alkylamino 1-3 alkyl; R 2 is acrylamidoin, A compound or a pharmaceutically acceptable salt thereof.

3. In paragraph 1, A 2 C is unsubstituted or substituted with one or more F 1-6 alkyl, A compound or a pharmaceutically acceptable salt thereof.

4. In paragraph 1, A 3 is Cl, A compound or a pharmaceutically acceptable salt thereof.

5. In paragraph 1, A 1 is methylcarbonyl or methylsulfonyl; A 2 is CF3CH2, CH3CH2, CF3CH2CH2 or CH3; A 3 is Cl; R 1 is -NR b1 R b2 And, R b1 Silver methyl; R b2 is 2-(dimethylamino)ethyl; R 2 is acrylamidoin, A compound or a pharmaceutically acceptable salt thereof.

6. In paragraph 1, the compound is selected from the following or a pharmaceutically acceptable salt thereof: <1> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide; <2> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(2,2,2-trifluoroethoxy)pyridin-3-yl)acrylamide; <3> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide; <4> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-ethoxypyridin-3-yl)acrylamide; <5> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide; <6> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-(3,3,3-trifluoropropoxy)pyridin-3-yl)acrylamide; <7> N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide; <8> N-(5-((5-chloro-4-((1-(methylsulfonyl)indolin-7-yl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-6-methoxypyridin-3-yl)acrylamide.

7. As shown in the following reaction formula 1, A method for producing a compound represented by chemical formula 1 of claim 1, comprising the step of producing a compound represented by chemical formula 1 by reacting a compound represented by chemical formula 2 with a compound represented by chemical formula 3: [Reaction Formula 1] A in the above reaction scheme 1 1 , A 2 , A 3 , R 1 and R 2 is as defined in Chemical Formula 1 of Article 1.

8. A pharmaceutical composition for preventing or treating cancer, containing a compound represented by Chemical Formula 1 of Article 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

9. A pharmaceutical composition characterized in that the compound represented by chemical formula 1 of paragraph 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof inhibits EGFR (epidermal growth factor receptor) mutation or HER2 mutation.

10. In paragraph 9, The above EGFR mutations may include del19, L858R, V769_D770insASV or D770_N771insSVD, A pharmaceutical composition characterized in that the above HER2 mutation may include A775_G776insYVMA.

11. In paragraph 8, The above cancers are pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, A pharmaceutical composition characterized in that it is at least one selected from the group consisting of duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid tumor, vaginal cancer, spinal cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsillar cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, and thymic cancer.

12. A health functional food composition for preventing or improving cancer, containing a compound represented by Chemical Formula 1 of Article 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

13. A method for treating cancer, comprising administering a therapeutically effective amount of a compound according to paragraph 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

14. Use of a compound according to paragraph 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer.

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