WO2021075691A1 - Pyrimidine derivative, method of preparing same, and pharmaceutical composition for preventing or treating cancer, comprising same as effective component - Google Patents

Abstract

The present invention relates to a pyrimidine derivative, a method of preparing same, and a pharmaceutical composition for preventing or treating cancer, comprising the pyrimidine derivative as an effective component, wherein the derivative exhibits a relatively weak inhibitory effect on EGFR activity with respect to wild-type EGFR while exhibiting a high inhibitory effect with respect to EGFR mutations, and thus can be effectively used for the treatment of cancer with EGFR mutations.

Description

Pyrimidine derivative, preparation method thereof, and pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient

The present invention relates to a pharmaceutical composition for preventing or treating cancer, and specifically to a pharmaceutical composition for preventing or treating cancer containing a pyrimidine derivative as an active ingredient.

Cancer incidence is related to changes in various environmental factors including chemicals, radiation, and viruses, oncogenes, tumor suppressor genes, and genes related to apoptosis and DNA repair. By understanding the mechanism, a new treatment, targeted chemotherapy, became possible.

Targeted therapies are generally made to target molecules characteristic of cancer cells to exert their effects, and molecular targets are cancer cell signal transduction pathways, angiogenesis, and interstitial cells. They are genes related to (matrix), cell cycle regulator, and apoptosis. Currently used as important target therapeutics in treatment include tyrosine kinase inhibitors, signaling pathway inhibitors, and angiogenesis inhibitors.

Protein tyrosine kinase has been shown to play an important role in many malignancies. In particular, epidermal growth factor receptor (EGFR), a receptor tyrosine kinase of the erbB family, is a non-small cell lung cancer. It is abnormally activated in many epithelial cell tumors, including 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. It has been known that activation causes continuous cell proliferation, invasion of surrounding tissues, distant metastasis, and blood vessel formation, and increases cell survival.

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

In particular, EGFR is overexpressed in more than half of non-small cell lung cancer (NSCLC), and many studies have been conducted as a target of treatment. EGFR TKI (tyrosine kinase inhibitor), which inhibits EGFR tyrosine kinase activity, has been developed, and representative drugs include gefitinib (IRESSA™), erlotinib (TARCEVA™), and lapatinib (TYKERB™, TYVERB™).

Meanwhile, in 2004, it was reported that the activating mutation of EGFR correlates with the response to gefitinib therapy in non-small-cell lung cancer (NSCLC) (Science [2004] Vol.304, 1497- 500 and New England Journal of Medicine [2004] Vol. 350, 2129-39).

Specifically, the EGFR mutation is largely classified into a sensitizing mutation and a resistant mutation. The deletion of exon 19 and the L858R point mutation of exon 21 are the most important sensitive mutations. %, and exon 19 del mutations are known to be more sensitive to TKI. On the other hand, the T790M point mutation of exon 20 is the most important resistance mutation and is known to be found in more than 50% of acquired resistance patients (Clin Cancer Res 2006;12:6494-6501.).

Somatic mutations identified so far include in-frame deletions within exon 19 or insertions in exon 20, as well as point mutations in which a single nucleic acid residue is modified in the expressed protein (e.g., L858R, G719S, G719C, G719A, L861Q) ( Fukuoka et al. JCO 2003; Kris et al JAMA 2003 and Shepherd et al NEJM 2004).

Despite the initial clinical effects of gefitinib/erlotinib in NSCLC patients with EGFR mutations, most patients eventually develop advanced cancer during therapy with these agents. In an initial study of relapsed specimens, a secondary EGFR mutation, T790M, was identified that renders gefitinib and erlotinib ineffective inhibitors of EGFR kinase activity (Kobayashi et al NEJM 2005 and Pao et al PLOS Medicine 2005). It was demonstrated in subsequent studies that the EGFR T790M mutation was found in approximately 50% (24/48) of tumors from patients who acquired resistance to gefitinib or erlotinib (Kosaka et al CCR 2006; Balak et al CCR 2006 and Engelman et al Science 2007). This secondary genetic modification results from a position similar to the'gatekeeper' residue and the secondary resistance allele associated with it in patients treated with kinase inhibitors (e.g., T315I in ABL in imatinib resistant CML).

It has long been known that EGFR mutations, EGFR_del19 or EGFR_L858R, are the main causes of non-small cell lung cancer and head and neck cancer, and their therapeutic drugs, Iressa and Taseba, have been developed and are currently used in clinical practice. However, when these drugs were used in patients, acquired resistance, which caused the EGFR secondary mutation based on the structure of the drug, was observed, and it was also found that this is the main cause of actual drug resistance. When the first generation EGFR inhibitor is used for an average of 10 months, the acquired resistance of the T790M mutation located at the gatekeeper of the EGFR kinase occurs, and the first generation EGFR inhibitors do not take effect. In other words, EGFR_del19_T790M or EGFR_L858R_T790M double mutation occurs, preventing the existing treatment from showing efficacy.

Based on these facts, the need for the development of 2nd and 3rd generation drugs with excellent drug efficacy and new structures is emerging.

In addition, there is no suitable therapeutic agent for rare EGFR mutations other than EGFR_del19 or EGFR_L858R mutations. In particular, there is no suitable therapeutic agent for EGFR Exon20insertion mutation. There is a wide variety of EGFR Exon20insertion mutations, including EGFR D770delinsGY, EGFR H773_V774insH, EGFR Y764_V765insHH, EGFR D770insASV and EGFR D770_N771insSVD.

Accordingly, there is a need for the development of inhibitors that exhibit relatively low inhibition against wild-type (WT) EGFR and higher inhibition against specific activated or resistant mutant forms of EGFR.

Accordingly, while trying to develop a cancer therapeutic agent that inhibits EGFR multiple mutations, the pyrimidine derivatives according to the present invention exhibit relatively low inhibition against wild-type EGFR, while exhibiting high inhibitory ability against EGFR mutations, thereby preventing cancer. Alternatively, the present invention was completed by finding that it can be usefully used for treatment.

It is an object of the present invention to provide a pyrimidine derivative, an isomer thereof, or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preparing the pyrimidine derivative.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer containing the pyrimidine derivative, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving cancer containing the pyrimidine derivative, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating diseases related to EGFR mutations containing the pyrimidine derivative, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

To achieve the above object,

According to an aspect of the present invention, there is provided a compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Formula 1,

R is -OR ^a or -NR ^b1 R ^b2 ,

R ^a is -(CH ₂ ) _m -NR ^b1 R ^b2 , where m is 1 to 3,

R ^b1 and R ^b2 are each independently hydrogen or unsubstituted or straight or branched chain C _1-6 ^{alkyl substituted with NR c1} R ^c2 , or R ^b1 and R ^b2 are N, O together with the nitrogen to which they are attached. _{And to form an unsubstituted or straight or branched chain C 1-6} alkyl containing one or more heteroatoms selected from the group consisting of S or a 5 to 7 membered heterocycloalkyl substituted with ^{NR c1} R ^c2,

R ^c1 and R ^c2 are each independently hydrogen or straight or branched chain C _1-6 alkyl, or, R ^c1 and R ^c2 are selected from the group consisting of N, O and S together with the nitrogen to which they are attached. 5 to 7 membered heterocycloalkyl substituted with unsubstituted or straight or branched C _{1-6 alkyl containing one or more atoms.}

According to another aspect of the present invention, as shown in Scheme 1 below,

It provides a method for preparing a compound represented by Formula 1, comprising the step of preparing a compound represented by Formula 1 by reacting a compound represented by Formula 2 with a compound represented by Formula 3:

[Scheme 1]

In Reaction Scheme 1, R is as defined in Chemical Formula 1.

According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating cancer containing the compound represented by Chemical Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

According to another aspect of the present invention, there is provided a health functional food for preventing or improving cancer containing the compound represented by Formula 1, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating diseases related to EGFR mutations containing the compound represented by Formula 1, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

According to another aspect of the present invention, cancer or EGFR comprising administering to a subject in need a pharmaceutical composition or a dietary supplement composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient It provides a method of preventing or treating mutation-related diseases.

According to another aspect of the present invention, use of a pharmaceutical composition or a dietary supplement composition containing a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in the prevention or treatment of cancer or EGFR mutation-related diseases Provides.

The pyrimidine derivative according to the present invention exhibits a relatively weak inhibitory effect on EGFR activity against wild-type EGFR, and exhibits a high inhibitory ability against EGFR mutations, so it can be usefully used in the treatment of cancer in which EGFR mutations have occurred.

1 is a graph showing the results of measuring the cell growth inhibitory ability of the existing EGFR TKI (tyrosine kinase inhibitor) drug osimertinib against the Ba/F3 EGFR exon20 insertion mutation.

2 is a graph showing the results of measuring the cell growth inhibitory ability of the compound represented by Example 1 against the Ba/F3 EGFR exon20 insertion mutation.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R is -OR ^a or -NR ^b1 R ^b2 ,

R ^a is -(CH ₂ ) _m -NR ^b1 R ^b2 , where m is 1 to 3,

R ^b1 and R ^b2 are each independently hydrogen or unsubstituted or straight or branched chain C _1-6 ^{alkyl substituted with NR c1} R ^c2 , or R ^b1 and R ^b2 are N, O together with the nitrogen to which they are attached. _{And to form an unsubstituted or straight or branched chain C 1-6} alkyl containing one or more heteroatoms selected from the group consisting of S or a 5 to 7 membered heterocycloalkyl substituted with ^{NR c1} R ^c2,

R ^c1 and R ^c2 are each independently hydrogen or straight or branched chain C _1-6 alkyl, or, R ^c1 and R ^c2 are selected from the group consisting of N, O and S together with the nitrogen to which they are attached. 5 to 7 membered heterocycloalkyl substituted with unsubstituted or straight or branched C _{1-6 alkyl containing one or more atoms.}

In Formula 1,

R is

,

,

,

,

,

,

,

or

Can be

Examples of the compound represented by Formula 1 according to the present invention include the following compounds:

<1> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino) Ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide;

<2> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(4-(dimethylamino)pi Peridin-1-yl)-4-methoxyphenyl)acrylamide;

<3> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(4- (4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide;

<4> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-morpholino Phenyl)acrylamide;

<5> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(4- Methylpiperazin-1-yl)phenyl)acrylamide;

<6> to N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(2-(dimethylamino) Oxy)-4-methoxyphenyl)acrylamide;

<7> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(methyl( 2-morpholinoethyl)amino)phenyl)acrylamide;

<8> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(methyl( 2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide; And

<9> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amido)pyrimidin-2-yl)amino)-4-methoxy-2-(2 -Morpholinoethoxy)phenyl)acrylamide.

The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. It is obtained from non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids, etc., acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, i. Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Rate, sebacate, fumarate, maleate, butine-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, 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, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate formed by dissolving the derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid It can be prepared by filtration and drying, or it can be prepared by distilling a solvent and an excess of acid under reduced pressure and then drying to crystallize under an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be made using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).

Furthermore, the present invention includes not only the compound represented by Formula 1 and its pharmaceutically acceptable salts, but also solvates, isomers, hydrates, etc. that may be prepared therefrom.

The term "solvate" refers to a compound of the present invention or a salt thereof containing a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Preferred solvents therefor include volatile, non-toxic, and/or suitable solvents for administration to humans. In this case, when the solvent is water, it is referred to as "hydrate".

The term "isomer" refers to a compound of the present invention or a salt thereof having the same chemical formula or molecular formula, but structurally or sterically different. Such isomers include structural isomers such as tautomers, R or S isomers having an asymmetric carbon center, stereoisomers such as geometric isomers (trans, cis), and optical isomers. All these isomers and mixtures thereof are also included within the scope of the present invention.

In addition, the present invention, as shown in the following Scheme 1,

It provides a method for preparing a compound represented by Formula 1, comprising the step of preparing a compound represented by Formula 1 by reacting a compound represented by Formula 2 with a compound represented by Formula 3.

[Scheme 1]

In Reaction Scheme 1, R is as defined in Chemical Formula 1.

Hereinafter, the manufacturing method represented by Scheme 1 will be described in detail.

In the manufacturing method represented by Scheme 1 according to the present invention, step 1 is a step of preparing a compound represented by Formula 1 by reacting a compound represented by Formula 2 with a compound represented by Formula 3. Specifically, this is a step in which the compound represented by Formula 1 is formed by reacting the halogen of the compound represented by Formula 2 with the primary amine of the compound represented by Formula 3.

The reaction of Scheme 1 is not particularly limited as long as it is a condition capable of forming an amine bond by combining a halogen and an amine.

In the present invention, an acid condition was used, and trifluoroacetic acid (TFA) was used as the acid, but the present invention is not limited thereto. In addition, the concentration of the acid may be 1 N, but is not limited thereto.

The solvent usable in Reaction Scheme 1 is not particularly limited, but lower alcohols including isopropanol, methanol, ethanol, propanol and butanol; Tetrahydrofuran (THF); Dioxane; Ether solvents including ethyl ether and 1,2-dimethoxyethane; Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), methylene chloride, dichloroethane, water, acetonase sulfonate, toluene sulfonate, chlorobenzene sulfonate, xylene sulfonate, ethyl acetate, phenyl acetate, phenyl propionate , Phenylbutyrate, Cycrate, lactate, hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, acetonite There are reels and the like, and these may be used alone or in combination. In the present invention, n-butanol, which is a lower alcohol, was used, but the present invention is not limited thereto.

In addition, the reaction temperature in Scheme 1 may be 80° C. to 95° C., but is not limited thereto, and the reaction temperature may be appropriately adjusted according to the degree to which the reaction proceeds.

In addition, the reaction time in Scheme 1 may be 10 minutes to 24 hours, but is not limited thereto, and the reaction time may be appropriately adjusted according to the degree to which the reaction proceeds.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating cancer containing the compound represented by Formula 1, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

At this time, the cancer is pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, cleft lip cancer, mycological sarcoma, acute myeloid leukemia, acute lymphocytic leukemia, basal cell Cancer, ovarian epithelial cancer, ovarian germ cell carcinoma, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, barter bulge cancer, bladder cancer, Peritoneal cancer, parathyroid cancer, adrenal cancer, non-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, kidney cancer, kidney cancer, 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, primary site unknown cancer, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal interstitial cancer , Wilms cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, pregnancy chorionic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, Auditory schwannoma, 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 It is one or more selected from the group consisting of thymic cancer, and the cancer may be a cancer in which a mutation is expressed for EGFR.

In this case, the EGFR mutations are EGFR del19, EGFR L858R, EGFR del19/T790M, EGFR L858R/T790M, EGFR del19/T790M/C797S, EGFR L858R/T790M/C797S, EGFR D770delinsGY, EGFR H773_V774ins D770ASH, EGFR EGFR and EGFR D770ASH, EGFR and EGFR D770ASH, EGFR, and EGFR del19/T790M_EGFR L858R/T790M It may be one or more selected from the group consisting of.

The compound represented by Formula 1 of the present invention exhibits a relatively weak inhibitory effect on EGFR activity against wild-type EGFR, while selectively exhibiting a high inhibitory ability against EGFR mutations, and in particular, exhibits a high inhibitory ability against EGFR del19/T790M, a double mutation. (See Experimental Example 1 and Table 2).

The compound represented by Formula 1 of the present invention exhibits a selectively high cell growth inhibitory ability against EGFR mutations in Ba/F3 cell lines.In particular, the existing EGFR against triple mutations EGFR del19/T790M/C797S or EGFR L858R/T790M/C797S Since it exhibits a higher cell growth inhibitory ability than Osimertinib, which is a tyrosine kinase inhibitor (TKI) drug, it can be seen that it exhibits superior anticancer effects than osimertinib (see Experimental Example 2, Table 3 and Table 4).

Example 1 compound of the present invention selectively shows high cell growth inhibitory ability against EGFR mutations in Ba/F3 cell lines, specifically EGFR D770delinsGY, EGFR H773_V774insH, EGFR Y764_V765insHH, EGFR D770insASV or EGFR D770_N771insSVD Since it exhibits a higher cell growth inhibitory ability than mertinib, it can be seen that it exhibits superior anticancer effects than osimertinib (see Experimental Example 3 and Table 5).

Therefore, since the compound represented by Formula 1 according to the present invention exhibits high inhibitory ability against EGFR mutations, EGFR del19, EGFR L858R, EGFR del19/T790M, EGFR L858R/T790M, EGFR del19/T790M/C797S, EGFR L858R/ T790M/C797S, EGFR D770delinsGY, EGFR H773_V774insH, EGFR Y764_V765insHH, EGFR D770insASV, EGFR D770_N771insSVD can be usefully used in the treatment of cancers expressing EGFR mutations. Since the inhibitory ability against T790M/C797S is remarkably excellent, it can be usefully used in the treatment of cancers expressing EGFR del19/T790M/C797S or EGFR L858R/T790M/C797S.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered in various oral and parenteral formulations upon clinical administration. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose, or lactose ( lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc., but may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are commonly used simple diluents. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, and emulsions. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.

A pharmaceutical composition containing the compound represented by 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. It depends on how to do it.

At this time, in order to formulate a formulation for parenteral administration, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed in water together with a stabilizer or buffer to prepare a solution or suspension, and the ampoule or vial unit dosage form It can be manufactured with. The composition may be sterilized and/or contain adjuvants such as preservatives, stabilizers, hydrating agents or emulsification accelerators, salts and/or buffers for controlling osmotic pressure, and other therapeutically useful substances, which are conventional methods of mixing and granulation. It can be formulated according to the method of painting or coating.

Formulations for oral administration include, for example, tablets, pills, hard/soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and troches. , Dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (such as silica, talc, stearic acid and magnesium or calcium salts thereof and/or polyethylene glycol). Tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and in some cases boron such as starch, agar, alginic acid or sodium salt thereof. It may contain release or boiling mixtures and/or absorbents, colorants, flavoring agents, and sweetening agents.

In addition, the present invention provides a health functional food for the prevention or improvement of cancer containing the compound represented by Chemical Formula 1, its isomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

The cancer is pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, cleft lip cancer, mycosis fungoides, acute myelogenous leukemia, acute lymphocytic leukemia, basal cell cancer, Ovarian epithelial cancer, ovarian germ cell carcinoma, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, barter bulge cancer, bladder cancer, peritoneal cancer , Parathyroid cancer, adrenal cancer, non-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 cancer, kidney cancer, 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, primary site unknown cancer, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal interstitial cancer, Wilm Cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational chorionic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, auditory nerve sheath , 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, hematologic cancer and thymic cancer It is one or more selected from the group consisting of, and the cancer may be a cancer in which a mutation is expressed for EGFR.

The compound represented by Formula 1 according to the present invention is a health functional food composition for the prevention or improvement of cancer, in particular, cancer in which EGFR mutation is expressed, by showing a high inhibitory ability against EGFR mutation. Can be added to.

The compound represented by Formula 1 according to the present invention may be added to food as it is or may be used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (for prevention or improvement). In general, the amount of the compound in the health food may be added in an amount of 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and there is no problem in terms of safety, so the active ingredient may be used in an amount above the above range.

In addition, the health functional beverage composition of the present invention is not particularly limited to other ingredients other than containing the compound as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates, etc. as additional ingredients, as in ordinary beverages. have. Examples of the above-described natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 g of the composition of the present invention.

Further, in addition to the above, the compound represented by Chemical Formula 1 according to the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavors and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.), pects. Acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, the compound represented by Chemical Formula 1 of the present invention may contain flesh for the manufacture of natural fruit juice and fruit juice beverages and vegetable beverages.

In addition, the present invention provides a pharmaceutical composition for preventing or treating diseases related to EGFR mutations containing the compound represented by Formula 1, isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient.

The EGFR mutation-related disease may be at least one selected from the group consisting of cancer, psoriasis, eczema, atherosclerosis, stomatous rash, dry skin, chronic diarrhea, liver cirrhosis, myocardial fibrosis, and chronic renal failure.

In addition, the present invention is a cancer or EGFR mutation-related disease comprising the step of administering to a subject in need a pharmaceutical composition or a dietary supplement composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient It provides a method of preventing or treating.

In addition, the present invention provides a use of a pharmaceutical composition or a health functional food composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in the prevention or treatment of cancer or EGFR mutation-related diseases. .

Hereinafter, the present invention will be described in detail by examples and experimental examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following Examples and Experimental Examples.

<Preparation Example 1-1> Preparation of N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)methanesulfonamide

N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)methanesulfonamide was prepared through the method represented by Scheme 2 below.

[Scheme 2]

Step 1: Preparation of 2,5-dichloro-N-(2-nitrophenyl)pyrimidin-4-amine

2-nitroaniline (10 g, 72.4 mmol) was dissolved in DMF (100 mL) and NaH (3.47 g, 144.8 mmol) was added to the stirred solution, followed by stirring at 0° C. for 30 minutes. Then 2,4,5-trichloropyrimidine (16.0 g, 86.8 mmol) was added at 0°C. The reaction mixture was quenched with ice water (300 mL) and stirred for 15 minutes. The precipitated solid was filtered and dried under vacuum to give 2,5-dichloro-N-(2-nitrophenyl)pyrimidin-4-amine (17 g, 85% yield, yellow solid).

¹ H NMR (300 MHz, DMSO- d ₆ ) δ 10.23 (s, 1 H), 8.49 (s, 1 H), 8.10 (dd, J = 8.2, 1.4 Hz, 1 H), 7.91-7.71 (m, 2H), 7.53-7.43 (m, 1H).

Step 2: Preparation of N-(2,5-dichloropyrimidin-4-yl)benzene-1,2-diamine

Dissolving 2,5-dichloro-N-(2-nitrophenyl)pyrimidin-4-amine (17 g, 59.6 mmol) prepared in step 1 in THF/H ₂ O (1:1, 300 mL) Iron powder (16.6 g, 298 mmol) and NH ₄ Cl (15.9 g, 298.1 mmol) were added to the stirred solution at room temperature. The reaction mixture was heated to 65° C. for 5 hours. This reaction was observed by TLC. The reaction mixture after heating was cooled to room temperature, filtered through Celite, and washed with ethyl acetate. The organic layer was separated, washed with brine, dried over Na ₂ SO ₄ , evaporated under reduced pressure and N-(2,5-dichloropyrimidin-4-yl)benzene-1,2-diamine (10 g, 65% Yield, yellow solid).

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 9.02 (s, 1 H), 8.26 (s, 1 H), 7.06-6.95 (m, 2 H), 6.77 (dd, J = 8.1, 1.4 Hz, 1H), 6.58 (td, J = 7.5, 1.4 Hz, 1H), 4.98 (s, 2H); LCMS: 256.8 [M+H ⁺ ].

Step 3: Preparation of N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)methanesulfonamide

Dissolve N-(2,5-dichloropyrimidin-4-yl)benzene-1,2-diamine (3 g, 11.5 mmol) prepared in step 2 in DCM (50 mL) and add Et _{3 to the stirred solution.} N (5 mL, 35.0 mmol) was added at room temperature. The reaction mixture was cooled to 0°C, and mesyl chloride (2.3 mL, 23.0 mmol) was added dropwise. After that, the reaction mixture was warmed to room temperature and left for 14 hours. This reaction was observed by TLC. The reaction mixture _{was quenched with saturated NaHCO 3} aqueous solution (30 mL), and extracted with DCM (30 mL). The organic layer was washed with water, brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure. The obtained crude was dissolved in methanol (50 mL), K ₂ CO ₃ (2.0 g) was added, and stirred for 12 hours. The stirred liquid was evaporated under reduced pressure. Subsequently, the obtained stock solution was _{diluted with H 2} O (100 mL) and extracted with DCM (100 mL). The aqueous layer was separated, acidified with 1N HCl (pH ~ 5-4), and extracted with DCM. The organic layer was separated _{, dried over Na 2} SO ₄ , concentrated under reduced pressure, and N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)methanesulfonamide (2.5 g, 64% yield) , A pale red solid) was obtained.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 10.03 (s, 1 H), 8.33 (s, 1 H), 8.24 (dd, J = 8.0, 1.5 Hz, 1 H), 7.19 (dd, J = 8.0, 1.3 Hz, 1H), 6.83 (td, J = 7.7, 1.6 Hz, 1H), 6.58 (td, J = 7.6, 1.4 Hz, 1H), 2.64 (s, 3H); LCMS: 334.8 [M+H ⁺ ].

<Production Example 1-2> Preparation of tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) carbamate

Tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) carbamate was prepared through the method represented by Scheme 3 below.

[Scheme 3]

Step 1: Preparation of 4-fluoro-2-methoxy-5-nitroaniline

To concentrated H ₂ SO ₄ (50 mL) 4-fluoro-2-methoxyaniline (4.7 g, 33.0 mmol) was added portionwise, cooled in an ice water bath, and the temperature was kept below 15°C. The mixture was stirred until the solid dissolved. Potassium nitrate (3.36 g, 33.0 mmol) was added portionwise, and the temperature was kept below 10°C. The mixture was then stirred overnight and poured onto ice/water. The mixture was then basified with concentrated NH ₄ OH. The solid was filtered, dissolved in DCM, washed with water, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The product stock solution was purified by flash silica chromatography, and was eluted with DCM containing 1 to 10% MeOH. The pure fractions were evaporated and dried to obtain 4-fluoro-2-methoxy-5-nitroaniline (4.5 g, 76% yield, yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.38-7.29 (d, J = 7.8 Hz, 1 H), 7.11-6.97 (d, J = 13.4 Hz, 1 H), 5.24 (s, 2 H) , 3.91 (s, 3H); LCMS: 187.2 [M+H ⁺ ].

Step 2: Preparation of tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) carbamate

4-fluoro-2-methoxy-5-nitroaniline (4.5 g, 24.1 mmol) and DMAP (300 mg, 2.41 mmol) prepared in step 1 were dissolved in DCM and added to the stirred solution (Boc) ₂ O (5.8 g, 26.2 mmol) was added at 0°C. The reaction mixture was slowly warmed to room temperature and left for 12 hours. Thereafter, the reaction mixture was diluted with DCM and quenched with _{saturated NaHCO 3 aqueous solution.} The separated organic layer was washed with water and brine, dried over Na ₂ SO ₄ , and concentrated under reduced pressure. The obtained stock solution was purified using column chromatography in which the eluent was hexanes/ethyl acetate (7:3), and tert-butyl (4-fluoro-2-methoxy -5-nitrophenyl)carbamate (4.0 g, 58% yield, yellow solid) was obtained.

¹ H NMR (400 MHz, Chloroform- d ) δ 8.91 (d, J = 8.0 Hz, 1 H), 6.99 (s, 1 H), 6.78-6.68 (d, J = 12.1 Hz, 1 H), 4.00 ( s, 3H), 1.56 (s, 9H); LCMS: 287.2 [M+H ⁺ ].

<Production Example 1-3> Preparation of N-(5-amino-2((2-(dimethylamino)ethyl(methyl)amino)-4-methoxyphenyl)acrylamide

N-(5-amino-2((2-(dimethylamino)ethyl(methyl)amino)-4-methoxyphenyl)acrylamide was prepared through the method represented by Scheme 4 below.

[Scheme 4]

Step 1: Preparation of tert-butyl (4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenyl)carbamate

The tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) carbamate (300 mg, 1.04 mmol) prepared in Preparation Example 1-2 was dissolved in DMF (5 mL) and added to the stirred solution. N ¹ ,N ¹ ,N ² -Trimethylethane-1,2-diamine (106 mg, 1.04 mmol) and cesium carbonate (183 mg, 1.56 mmol) were added at room temperature. The reaction mixture was stirred at 70° C. overnight. Thereafter, the reaction mixture was cooled to room temperature and quenched with ice water. The precipitated solid was filtered and dried under vacuum, tert-butyl (4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenyl)carbamate (250 mg, 65% yield, yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.16 (s, 1 H), 8.11 (s, 1 H), 6.74 (s, 1 H), 3.90 (s, 3 H), 3.24-3.19 (t , J = 6.8 Hz, 2H), 2.80 (s, 3H), 2.46-2.40 (t, J = 6.8 Hz, 2H), 2.13 (s, 6H), 1.45 (s, 9H); LCMS: 369.0 [M+H ⁺ ].

Step 2: Preparation of tert-butyl(5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenyl)carbamate

Tert-butyl (4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenyl)carbamate (250 mg, 0.64 mmol) prepared in step 1 was added to methanol ( 15 mL), 10% Pd/C (6.8 mg, 0.06 mmol) was added to the stirred solution under an argon (Ar, Argon) atmosphere. The reaction mixture was stirred at room temperature for 4 hours under 1 atm hydrogen gas pressure, and concentrated under reduced pressure, tert-butyl(5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2- Methoxyphenyl) carbamate (210 mg, 91% yield, brown solid) was obtained.

LCMS: 339.2 [M+H ⁺ ]

Step 3: Preparation of tert-butyl (5-acrylamido-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenyl)carbamate

Tert-butyl (5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenyl)carbamate (200 mg, 0.59 mmol) and triethyl prepared in step 2 Amine (0.16 mL, 1.18 mmol) was dissolved in DCM, and acryloyl chloride (0.047 mL, 0.59 mmol) was added to the stirred solution at -30°C. The reaction mixture was slowly warmed to room temperature and left for 3 hours. Thereafter, the reaction mixture was diluted with DCM and quenched with _{saturated NaHCO 3 aqueous solution.} The organic layer was separated, washed with water and brine, dried over Na ₂ SO ₄ , and concentrated under reduced pressure. The obtained stock solution was purified using column chromatography in which the eluent was DCM/MeOH (9.5:0.5), and tert-butyl (5-acrylamido-4-((2-(dimethylamino)ethyl)(methyl)amino )-2-methoxyphenyl)carbamate (150 mg, 65% yield, off white solid) was obtained.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.03 (s, 1 H), 8.47 (s, 1 H), 7.83 (s, 1 H), 6.93 (s, 1 H), 6.49-6.34 (m , 1 H), 6.27-6.18 (dd, J = 17.0, 2.2 Hz, 1 H), 5.78-5.69 (dd, J = 10.0, 2.2 Hz, 1 H), 3.79 (s, 3 H), 2.91-2.82 (t, J = 5.9 Hz, 2H), 2.66 (s, 3H), 2.33 (bs, 2H), 2.22 (s, 6H), 1.45 (s, 9H); LCMS: 393.0 [M+H ⁺ ].

Step 4: Preparation of N-(5-amino-2((2-(dimethylamino)ethyl(methyl)amino)-4-methoxyphenyl)acrylamide

Tert-butyl (5-acrylamido-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenyl)carbamate (150.0 mg, 0.38 mmol) prepared in step 3 25% TFA dissolved in DCM (1 mL) was added to the stirred solution in DCM at room temperature. The mixture was stirred at room temperature for 12 hours. The reaction mixture was observed by TLC. Thereafter, N-(5-amino-2((2-(dimethylamino)ethyl(methyl)amino)-4-methoxyphenyl)acrylamide as a stock solution was concentrated under reduced pressure, and there was no other purification.

LCMS: 293.2 [M+H ⁺ ].

<Preparation Example 2-1> Preparation of tert-butyl (5-acrylamido-4-(4-(dimethylamino) piperidin-1-yl)-2-methoxyphenyl) carbamate

[tert-butyl (5-acrylamido-4-(4-(dimethylamino) piperidin-1-yl)-2-methoxyphenyl) carbamate]

According to steps 1 to 3 of Preparation Example 1-3, except that N,N-dimethylpiperidin-4-amine was used instead of ^{N 1} ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} tert-butyl (5-acrylamido-4-(4-(dimethylamino)piperidin-1-yl)-2-methoxyphenyl)carbamate (78% yield) was prepared.

* ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.95 (s, 1 H), 8.16 (s, 1 H), 7.83 (s, 1 H), 6.76 (s, 1 H), 6.69-6.57 ( m, 1 H), 6.26-6.17 (dd, J = 16.9, 2.0 Hz, 1 H), 5.78-5.67 (m, 1 H), 3.79 (s, 3 H), 3.05-2.96 (m, 2 H) , 2.69-2.57 (m, 2H), 2.28 (s, 7H), 1.88-1.79 (m, 2H), 1.76-1.61 (m, 2H), 1.45 (s, 9H); LCMS: 419.0 [M+H ⁺ ].

<Production Example 2-2> Preparation of N-(5-amino-2-(4-(dimethylamino)piperidin-1-yl)-4-methoxyphenyl)acrylamide

[N-(5-amino-2-(4-(dimethylamino)piperidin-1-yl)-4-methoxyphenyl)acrylamide]

Using tert-butyl (5-acrylamido-4-(4-(dimethylamino) piperidin-1-yl)-2-methoxyphenyl) carbamate prepared in Preparation Example 2-1 as a precursor Except, N-(5-amino-2-(4-(dimethylamino)piperidin-1-yl)-4-methoxyphenyl)acrylamide was prepared according to Step 4 of Preparation Example 1-3. .

LCMS: 319.0 [M+H ⁺ ].

<Production Example 3-1> tert-butyl (5-acrylamido-2-methoxy-4-(4-(4-methylpiperazin-1-yl) piperidin-1-yl) phenyl) carbamate Manufacture of

[tert-butyl (5-acrylamido-2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)carbamate]

The steps of Preparation Example 1-3, except that 1-methyl-4-(piperidin-4-yl)piperazine was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} Tert-butyl (5-acrylamido-2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)carbamate (65 % Yield).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.94 (s, 1H), 8.16 (s, 1 H), 7.85 (s, 1 H), 6.75 (s, 1 H), 6.71-6.57 (m, 1 H), 6.27-6.16 (dd, J = 16.9, 2.1 Hz, 1 H), 5.75-5.67 (dd, J = 10.3, 2.0 Hz, 1 H), 3.79 (s, 3 H), 3.06-2.95 ( d, J = 11.4 Hz, 2H), 2.91-2.58 (m, 10H), 1.93-1.78 (bs, 2H), 1.77-1.62 (bs, 2H), 1.45 (s, 9H); LCMS: 474.2 [M+H ⁺ ].

<Production Example 3-2> Preparation of N-(5-amino-4-methoxy-2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide

[N-(5-amino-4-methoxy-2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide]

Tert-butyl prepared in Preparation Example 3-1 as a precursor (5-acrylamido-2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl) N-(5-amino-4-methoxy-2-(4-(4-methylpiperazin-1-yl)pi according to Step 4 of Preparation Example 1-3 above, except that phenyl) carbamate was used. Peridine-1-yl)phenyl)acrylamide was prepared and used in the later step without purification.

<Production Example 4-1> Preparation of tert-butyl (5-acrylamido-2-methoxy-4-morpholinophenyl) carbamate

[tert-butyl (5-acrylamido-2-methoxy-4-morpholinophenyl) carbamate]

Tert-butyl (5-acrylamido-) according to steps 1 to 3 of Preparation Example 1-3, except that morpholine was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} 2-methoxy-4-morpholinophenyl)carbamate (77% yield) was prepared.

¹ H NMR (400 MHz, Chloroform- d ) δ 6.99 (s, 1 H), 6.73 (s, 1 H), 6.51-6.42 (d, J = 16.5 Hz, 1 H), 6.27-6.17 (m, 1 H), 5.82-5.74 (m, 1H), 3.99 (bs, 4H), 3.89 (s, 3H), 2.12-2.93 (bs, 4H), 1.54 (s, 9H); LCMS: 378.0 [M+H ⁺ ].

<Production Example 4-2> Preparation of N-(5-amino-4-methoxy-2-morpholinophenyl)acrylamide

[N-(5-amino-4-methoxy-2-morpholinophenyl)acrylamide]

Step 4 of Preparation Example 1-3, except that tert-butyl (5-acrylamido-2-methoxy-4-morpholinophenyl) carbamate prepared in Preparation Example 4-1 was used as a precursor. According to the following, N-(5-amino-4-methoxy-2-morpholinophenyl)acrylamide was prepared.

LCMS: 278.0.2 [M+H ⁺ ].

<Preparation Example 5-1> Preparation of tert-butyl (5-acrylamido-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)carbamate

[tert-butyl (5-acrylamido-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)carbamate]

Tert-butyl (5-acrylic) according to steps 1 to 3 of Preparation Example 1-3, except that 1-methylpiperazine was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine.} Amido-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)carbamate (77% yield) was prepared.

¹ H NMR (400 MHz, Chloroform- d ) δ 9.09 (s, 1 H), 8.31 (s, 1 H), 6.94 (s, 1 H), 6.74 (s, 1 H), 6.49-6.35 (d, J = 16.8 Hz, 1 H), 6.33-6.22 (dd, J = 17.4, 10.0 Hz, 1 H), 5.80-5.68 (d, J = 10.2 Hz, 1 H), 3.85 (s, 3 H), 3.03 (bs, 4H), 2.96-2.77 (m, 4H), 2.55 (s, 3H), 1.54 (s, 9H); LCMS: 391.0 [M+H ⁺ ].

<Production Example 5-2> Preparation of N-(5-amino-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)acrylamide

[N-(5-amino-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)acrylamide]

Except for using tert-butyl (5-acrylamido-2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) carbamate prepared in Preparation Example 5-1 as a precursor, the above According to Step 4 of Preparation Example 1-3, N-(5-amino-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)acrylamide was prepared.

LCMS: 291.2 [M+H ⁺ ].

<Preparation Example 6-1> Preparation of tert-butyl (5-acrylamido-4-(2-(dimethylamino)ethoxy)-2-methoxyphenyl)carbamate

[tert-butyl (5-acrylamido-4-(2-(dimethylamino)ethoxy)-2-methoxyphenyl)carbamate]

Tert according to steps 1 to 3 of Preparation Example 1-3, except that 2-(dimethylamino)ethan-1-ol was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} -Butyl (5-acrylamido-4-(2-(dimethylamino)ethoxy)-2-methoxyphenyl)carbamate (71% yield) was prepared.

¹ H NMR (300 MHz, Chloroform- d ) δ 9.75 (s, 1 H), 9.00 (s, 1 H), 6.83 (s, 1 H), 6.57 (s, 1 H), 6.48-6.39 (dd, J = 16.9, 1.9 Hz, 1 H), 6.34-6.21 (dd, J = 16.9, 10.0 Hz, 1 H), 5.71-5.65 (dd, J = 10.0, 1.9 Hz, 1 H), 4.14-4.07 (m , 2H), 3.83 (s, 3H), 2.65-2.55 (m, 3H), 2.37 (s, 6H), 1.53 (s, 12H); LCMS: 380.2 [M+H ⁺ ].

<Production Example 6-2> Preparation of N-(5-amino-2-(2-(dimethylamino)ethoxy)-4-methoxyphenyl)acrylamide

[N-(5-amino-2-(2-(dimethylamino)ethoxy)-4-methoxyphenyl)acrylamide]

Preparation above except that tert-butyl (5-acrylamido-4-(2-(dimethylamino)ethoxy)-2-methoxyphenyl)carbamate prepared in Preparation Example 6-1 was used as a precursor. N-(5-amino-2-(2-(dimethylamino)ethoxy)-4-methoxyphenyl)acrylamide was prepared according to step 4 of Example 1-3, and used in the subsequent step without purification.

<Production Example 7-1> Preparation of tert-butyl (5-acrylamido-2-methoxy-4- (2-morpholinoethoxy) phenyl) carbamate

[tert-butyl (5-acrylamido-2-methoxy-4-(2-morpholinoethoxy)phenyl) carbamate]

Tert- according to steps 1 to 3 of Preparation Example 1-3, except that 2-morpholinoethan-1-ol was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} Butyl (5-acrylamido-2-methoxy-4- (2-morpholinoethoxy) phenyl) carbamate (79% yield) was prepared.

¹ H NMR (300 MHz, Chloroform- d ) δ 8.88 (s, 1 H), 6.80 (s, 1 H), 6.51 (s, 1 H), 6.49-6.39 (d, J = 15.9 Hz, 1 H) , 6.27-6.17 (m, 1H), 5.81-5.67 (d, J = 11.8 Hz, 1H), 4.24 (s, 2H), 3.90 (bs, 4H), 3.84 (s, 3H), 3.09 -2.55 (bs, 6H), 1.53 (s, 9H); LCMS: 422.0 [M+H ⁺ ].

<Production Example 7-2> Preparation of N-(5-amino-4-methoxy-2-(2-morpholinoethoxy)phenyl)acrylamide

[N-(5-amino-4-methoxy-2-(2-morpholinoethoxy)phenyl)acrylamide]

*

Preparation Example, except that tert-butyl (5-acrylamido-2-methoxy-4- (2-morpholinoethoxy) phenyl) carbamate prepared in Preparation Example 7-1 was used as a precursor. N-(5-amino-4-methoxy-2-(2-morpholinoethoxy)phenyl)acrylamide was prepared according to Step 4 of 1-3.

LCMS: 322.2 [M+H ⁺ ].

<Preparation Example 8-1> Preparation of tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)carbamate

[tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)carbamate]

The above Preparation Example, except that N-methyl-2-(4-methylpiperazin-1-yl)ethan-1-amine was used instead of ^{N 1} ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} Tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)carba according to steps 1 to 3 of 1-3 Mate (70% yield) was prepared.

¹ H NMR (300 MHz, Chloroform- d ) δ 9.15 (s, 1 H), 9.02 (s, 1 H), 6.94 (s, 1 H), 6.70 (s, 1 H), 6.52-6.45 (d, J = 3.4 Hz, 1 H), 6.24-6.08 (m, 1 H), 5.81-5.70 (m, 1 H), 3.85 (s, 3 H), 3.06-2.98 (m, 2 H), 2.89-2.69 (m, 8H), 2.65 (s, 3H), 2.53 (s, 2H), 2.50 (s, 3H), 1.54 (s, 9H); LCMS: 447.0 [M+H ⁺ ].

<Preparation Example 8-2> Preparation of N-(5-amino-4-methoxy-2-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide

[N-(5-amino-4-methoxy-2-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide]

Tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl) prepared in Preparation Example 8-1 as a precursor N-(5-amino-4-methoxy-2-(methyl(2-(4-methylpiperazin-1-yl)ethyl) according to Step 4 of Preparation Example 1-3 above, except that carbamate was used. )Amino)phenyl)acrylamide was prepared.

LCMS: 348.2 [M+H ⁺ ].

<Preparation Example 9-1> Preparation of tert-butyl (5-acrylamido-2-methoxy-4- (methyl (2-morpholinoethyl) amino) phenyl) carbamate

[tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-morpholinoethyl)amino)phenyl)carbamate]

Steps 1 to 3 of Preparation Example 1-3, except that N-methyl-2-morpholinoethan-1-amine was used instead of N ¹ ,N ¹ ,N ^{2 -trimethylethane-1,2-diamine} According to the following, tert-butyl (5-acrylamido-2-methoxy-4-(methyl(2-morpholinoethyl)amino)phenyl)carbamate (69% yield) was prepared.

¹ H NMR (300 MHz, Chloroform- d ) δ 9.15 (s, 1 H), 9.12 (s, 1 H), 6.94 (s, 1 H), 6.52-6.42 (m, 1 H), 6.25-6.07 ( m, 1 H), 5.74 (d, J = 11.8 Hz, 1 H), 3.84 (s, 3 H), 3.81-3.71 (t, J = 5.9 Hz, 4 H), 3.41 (s, 3 H), 3.04 (s, 2H), 2.74-2.65 (m, 2H), 2.64 (s, 3H), 2.45 (bs, 4H), 1.54 (s, 9H); LCMS: 435.2 [M+H ⁺ ].

<Preparation Example 9-2> Preparation of N-(5-amino-4-methoxy-2-(methyl(2-morpholinoethyl)amino)phenyl)acrylamide

[N-(5-amino-4-methoxy-2-(methyl(2-morpholinoethyl)amino)phenyl)acrylamide]

Except for using tert-butyl (5-acrylamido-2-methoxy-4- (methyl (2-morpholinoethyl) amino) phenyl) carbamate prepared in Preparation Example 9-1 as a precursor, According to Step 4 of Preparation Example 1-3, N-(5-amino-4-methoxy-2-(methyl(2-morpholinoethyl)amino)phenyl)acrylamide was prepared.

LCMS: 335.2 [M+H ⁺ ].

<Example 1> N-5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino-2-((2-(dimethylamino) Preparation of ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino-2-((2- (Dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide was prepared in the same manner as shown in Scheme 5 below.

[Scheme 5]

N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)methanesulfonamide (100 mg, 0.3 mmol) prepared in Preparation Example 1-1 was added to n-butanol containing 1N TFA. N-(5-amino-2((2-(dimethylamino)ethyl(methyl)amino)-4-methoxyphenyl)acrylic prepared in Preparation Example 1-3 above in a solution dissolved in (30 mL) and stirred Amide (71.5 mg, 0.3 mmol) was added at room temperature The reaction tube was sealed with a Teflon-lined cap, and the reaction mixture was stirred overnight at 90° C. The stirring solution was concentrated under reduced pressure. Was dissolved in DCM, washed with saturated NaHCO ₃ aqueous solution, water and brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure The obtained stock solution was subjected to column chromatography (DCM:MeOH/9:1 to 8.5:1.5 ) Purified with N-5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino-2-((2-(dimethylamino) Ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide (40 mg, 23% yield, off-white solid) was obtained.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.03 (s, 1H), 9.29 (s, 1 H), 8.53 (s, 1 H), 8.39 (s, 1 H), 8.16 (s, 1 H ), 8.12 (s, 1 H), 8.07-8.01 (m, 1 H), 7.431-7.22 (m, 1 H), 7.06-7.04 (dd, J = 6.6, 3.1 Hz, 2 H), 6.96 (s , 1 H), 6.42-6.32 (m, 1 H), 6.23-6.19 (dd, J = 16.9, 2.1 Hz, 1 H), 5.76-5.73 (dd, J = 10.1, 2.1 Hz, 1 H), 3.76 (s, 3H), 2.94 (s, 3H), 2.91-2.88 (t, J = 5.8 Hz, 2H), 2.70 (s, 3H), 2.38 (s, 3H), 2.26 (s, 6 H); LCMS: 590.0 [M+H ⁺ ].

<Example 2> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(4-(dimethylamino )Piperidin-1-yl)-4-methoxyphenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(4 -(Dimethylamino)piperidin-1-yl)-4-methoxyphenyl)acrylamide, prepared in Preparation Example 2-2, N-(5-amino-2-(4-(dimethylamino)pi) It was prepared according to Example 1, except that peridin-1-yl)-4-methoxyphenyl)acrylamide was used (yield 27%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.97 (s, 1 H), 8.58 (s, 1 H), 8.17-8.08 (m, 3 H), 8.08-7.99 (m, 1 H), 7.33 -7.26 (dd, J = 7.3, 2.2 Hz, 1 H), 7.13-7.01 (m, 2 H), 6.79 (s, 1 H), 6.72-6.61 (dd, J = 16.9, 10.2 Hz, 1 H) , 6.25-6.14 (dd, J = 17.0, 2.1 Hz, 1 H), 5.77-5.67 (dd, J = 10.0, 2.0 Hz, 1 H), 3.76 (s, 3 H), 3.10-3.00 (m, 2 H), 2.74-2.59 (m, 2H), 2.31 (s, 7H), 1.91-1.80 (d, J = 11.9 Hz, 2H), 1.78-1.63 (m, 2H); LCMS: 615.2 [M+H ⁺ ].

<Example 3> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-( Preparation of 4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy -2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide was prepared in Preparation Example 3-2. It was prepared according to Example 1, except that oxy-2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide was used (yield 30%).

¹ H NMR (400 MHz, Methanol- d ₄ ) δ 8.11 (s, 1 H), 7.95 (s, 1 H), 7.83-7.77 (d, J = 7.4 Hz, 1 H), 7.49-7.43 (d, J = 7.5 Hz, 1 H), 7.35-7.26 (m, 2 H), 6.93 (s, 1 H), 6.61-6.51 (dd, J = 17.0, 10.2 Hz, 1 H), 6.43-6.36 (d, J = 16.9 Hz, 1 H), 5.89-5.83 (d, J = 10.3 Hz, 1 H), 3.89 (s, 3 H), 3.30-3.10 (m, 5 H), 2.99 (s, 3 H), 2.89 (s, 6H), 2.15-2.07 (d, J = 12.2 Hz, 2H), 1.97-1.82 (m, 2H); LCMS: 670.2 [M+H ⁺ ].

<Example 4> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-mo Preparation of polynophenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy -2-morpholinophenyl)acrylamide, except that the N-(5-amino-4-methoxy-2-morpholinophenyl)acrylamide prepared in Preparation Example 4-2 was used. It was prepared according to Example 1 (yield 31%).

¹ H NMR (400 MHz, Methanol- d ₄ ) δ 8.10 (s, 1H), 7.93 (s, 1 H), 7.82-7.74 (d, J = 7.8 Hz, 1 H), 7.49-7.43 (d, J = 7.6 Hz, 1 H), 7.39-7.26 (m, 2 H), 6.92 (s, 1 H), 6.61-6.51 (dd, J = 17.0, 10.1 Hz, 1 H), 6.42-6.32 (d, J = 17.0 Hz, 1 H), 5.89-5.81 (d, J = 10.4 Hz, 1 H), 3.89 (s, 7 H), 3.00 (s, 3 H), 2.98-2.91 (t, J = 4.6 Hz, 4H); LCMS: 575.2 [M+H ⁺ ].

<Example 5> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-( Preparation of 4-methylpiperazin-1-yl)phenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy -2-(4-methylpiperazin-1-yl)phenyl)acrylamide, prepared in Preparation Example 5-2, N-(5-amino-4-methoxy-2-(4-methylpiperazine-) It was prepared according to Example 1, except that 1-yl)phenyl)acrylamide was used (yield 41%).

¹ H NMR (400 MHz, Methanol- d ₄ ) δ 8.13 (s, 1 H), 7.85-7.78 (m, 1 H), 7.48-7.40 (m, 1 H), 7.33-7.24 m, 1 H), 6.89 (s, 1 H), 6.63-6.52 (dd, J = 17.0, 10.3 Hz, 1 H), 6.43-6.33 (d, J = 16.9 Hz, 1 H), 5.90-5.84 (d, J = 10.0 Hz , 1 H), 3.91 (s, 3 H), 3.67-3.58 (d, J = 11.9 Hz, 2 H), 3.30-3.21 (m, 4 H), 3.18-3.10 (d, J = 12.0 Hz, 2 H), 3.01 (s, 3H), 2.98 (s, 3H); LCMS: 587.2 [M+H ⁺ ].

<Example 6> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(2-(dimethylamino )Ethoxy)-4-methoxyphenyl)acrylamide preparation

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(2 -(Dimethylamino)ethoxy)-4-methoxyphenyl)acrylamide, prepared in Preparation Example 6-2, N-(5-amino-2-(2-(dimethylamino)ethoxy)-4- It was prepared according to Example 1 except for methoxyphenyl)acrylamide (yield 29%).

¹ H NMR (400 MHz, Methanol- d ₄ ) δ 8.11 (s, 1 H), 7.90 (s, 1 H), 7.87-7.79 (m, 1 H), 7.47-7.40 (d, J = 7.5 Hz, 1 H), 7.31-7.21 (m, 2 H), 6.87 (s, 1 H), 6.58-6.39 (m, 2 H), 5.93-5.85 (d, J = 9.8 Hz, 1 H), 4.56-4.58 (t, J = 5.0 Hz, 2 H), 3.95 (s, 3 H), 3.65-3.58 (t, J = 4.8 Hz, 2 H), 3.02 (s, 6 H), 2.98 (s, 3 H) ; LCMS: 576.8 [M+H ⁺ ].

<Example 7> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-( Preparation of methyl (2-morpholinoethyl) amino) phenyl) acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy -2-(methyl(2-morpholinoethyl)amino)phenyl)acrylamide was prepared in Preparation Example 7-2 above. N-(5-amino-4-methoxy-2-(2-morpholino) It was prepared according to Example 1, except that ethoxy)phenyl)acrylamide was used (yield 22%).

¹ H NMR (400 MHz, Methanol- d ₄ ) δ 8.13 (s, 1 H), 7.83-7.75 (m, 2 H), 7.49-7.41 (m, 1 H), 7.33-7.24 (m, 2 H) , 6.88 (s, 1 H), 6.60-6.49 (dd, J = 17.0, 9.9 Hz, 1 H), 6.48-6.40 (d, J = 16.8 Hz, 1 H), 5.95-5.88(d, J = 9.9 Hz, 1 H), 4.61-4.54 (t, J = 5.1 Hz, 2 H), 4.03 (bs, 4 H), 3.95 (s, 3 H), 3.67-3.60 (t, J = 4.9 Hz, 2 H ), 3.33 (bs, 4H), 2.97 (s, 3H); LCMS: 619.2 [M+2H ⁺ ].

<Example 8> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-( Preparation of methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy -2-(Methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide was prepared in Preparation Example 8-2 above N-(5-amino-4-methoxy- It was prepared according to Example 1, except that 2-(methyl(2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide was used (yield 27%).

¹ H NMR (500 MHz, Methanol- d ₄ ) δ 8.16 (s, 1 H), 7.88 (s, 1 H), 7.82-7.76 (d, J = 7.0 Hz, 1 H), 7.50-7.44 (m, 1 H), 7.34-7.27 (m, 2 H), 7.00 (s, 1 H), 6.64-6.55 (dd, J = 16.9, 10.0 Hz, 1 H), 6.52-6.46 (d, J = 17.0 Hz, 1 H), 5.99 (d, J = 10.1 Hz), 3.96 (s, 3 H), 3.51-3.45 (bs, 2 H), 3.41-3.35 (bs, 4 H), 3.23-3.14 (bs, 4 H) ), 3.08 (s, 2H), 2.97 (s, 3H), 2.87 (s, 3H), 2.82 (s, 3H); LCMS: 645.0 [M+2H ⁺ ].

<Example 9> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amido)pyrimidin-2-yl)amino)-4-methoxy-2- Preparation of (2-morpholinoethoxy)phenyl)acrylamide

The pyrimidine derivative compound according to the present invention, N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amido)pyrimidin-2-yl)amino)-4-me N-(5-amino-4-methoxy-2-(methyl(2-morpholino) prepared in Preparation Example 9-2) of oxy-2-(2-morpholinoethoxy)phenyl)acrylamide It was prepared according to Example 1, except that ethyl) amino) phenyl) acrylamide was used (yield 26%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.34-9.21 (m, 2 H), 8.48 (s, 1 H), 8.22 (s, 1 H), 8.17-8.09 (m, 2 H), 8.06 -7.98 (d, J = 7.9 Hz, 1 H), 7.36-7.27 (dd, J = 7.7, 1.8 Hz, 1 H), 7.16-7.02 (m, 2 H), 6.92 (s, 1 H), 6.67 -6.54 (m, 1 H), 6.26-6.15 (dd, J = 17.0, 2.0 Hz, 1 H), 5.80-5.69 (d, J = 12.2 Hz, 1 H), 3.77 (s, 3 H), 3.56 (t, J = 4.7 Hz, 4H), 2.02-2.90 (m, 5H), 2.68 (s, 3H), 2.43-2.28 (m, 6H); LCMS: 632.2 [M+H ⁺ ].

The chemical structural formulas of the compounds prepared in Examples 1 to 9 are summarized and shown in Table 1 below.

Example	Chemical structure	Example	Chemical structure
One		2
3		4
5		6
7		8
9

<Experimental Example 1> Wild-type EGFR and EGFR mutant enzyme inhibitory ability

In order to confirm the inhibitory ability of the compound represented by Formula 1 according to the present invention against wild-type EGFR and EGFR mutant enzymes, the following experiment was performed. The results are shown in Table 2 below.

The activity measurement for wild-type and EGFR mutant enzymes for the compounds of the present invention was tested as follows using the HTRF system sold by Cisbio. EGFR wild-type and EGFR del19/T790M mutant enzymes were used by purchasing a recombinant protein provided by Carna Biosciences.

The composition of the assay buffer used for the activity measurement was 50 mM Tris-HCl pH 7.5, 100 mM NaCl, 7.5 mM MgCl ₂ , 3 mM KCl, 0.01% Tween 20, 0.1% BSA, and 1 mM DTT. Here, the enzyme reaction was carried out using a peptide substrate labeled with 50 mM ATP and 0.5 mM biotin. The analysis of the EGFR activity inhibitory effect of the compound was performed according to the following analytical reaction recipe.

Component 1: 4 mL of EGFR wild-type or mutant enzyme

Component 2: 2 mL of compound solution

Component 3: 4 mL ATP and biotin-labeled peptide

Enzymatic reaction starts by mixing component 1 and component 2 first, and then adding component 3 to it. After 2 hours reaction at 37℃, 10 mL of a measurement solution consisting of streptavidin-XL665 and europium-labeled anti-phosphotyrosine antibody provided by Cisbio was added to the enzyme reaction solution and reacted at room temperature for 1 hour. Finally, the ratio of fluorescence values at 615 nm and 665 nm is calculated using Perkin-Elmer's Envision equipment, and the enzyme activity is quantitatively measured and the inhibitory ability of the compound is confirmed. The measured values measured at the concentrations of the seven compounds were analyzed using the Prism program (version 5.01, Graphpad Software, Inc.), and the IC ₅₀ value, an index of the inhibitory ability of the compound, was calculated.

	EGFR wt. IC 50 (μM)	EGFR del19/T790M IC 50 (μM)
Osimertinib	0.0031	<0.001
Example 1	<0.001	<0.001
Example 2	0.064	<0.001
Example 3	0.01	<0.001
Example 4	0.37	<0.001
Example 5	0.018	<0.001
Example 6	0.0016	<0.001
Example 7	0.0094	<0.001
Example 8	0.029	<0.001
Example 9	0.00065	<0.001

As shown in Table 4 above,

All the compounds of the present invention exhibit a relatively weak EGFR activity inhibitory effect against wild-type EGFR, and selectively exhibit high inhibitory ability against EGFR mutations, which is similar to the case of osimertinib, a conventional EGFR tyrosine kinase inhibitor (TKI) drug. Similar things were confirmed.

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

<Experimental Example 2> Measurement of wild-type Ba/F3 EGFR and Ba/F3 EGFR mutant cell growth inhibitory ability in Ba/F3 cell lines

In order to confirm the ability of the compound represented by Formula 1 according to the present invention to inhibit cell growth against wild-type Ba/F3 EGFR and Ba/F3 EGFR mutations in Ba/F3 cell lines, the following experiment was performed. The results are shown in Tables 3 and 4 below.

* The activity of the compounds of the present invention against wild-type and mutant Ba/F3 EGFR cell lines was tested as follows using the CellTiter-Glo system sold by Promega. CellTiter-Glo assay is a method to check cell viability by measuring ATP present in cells in the cell culture state. Ba/F3 EGFR wild type and Ba/F3 EGFR del19, Ba/F3 EGFR L858R, Ba/F3 EGFR del19/T790M, Ba/F3 EGFR L858R/T790M, Ba/F3 del19/T790M/C797S, Ba/F3 EGFR L858R/T790M The /C797S mutant cell line was used by purchasing a cell line provided by Crown Bioscience. Ba/F3 EGFR wild type and Ba/F3 EGFR del19, Ba/F3 EGFR L858R, Ba/F3 EGFR del19/T790M, Ba/F3 EGFR L858R/T790M, Ba/F3 del19/T790M/C797S, Ba/F3 EGFR L858R/T790M The /C797S mutant cell line was cultured in an RPMI containing 10% FBS and 1% penicillin-streptomycin with 1 ug of puromycine in an incubator at _{37°C and 5% CO 2.}

Analysis of the EGFR cell growth inhibitory effect of the compound was carried out according to the following analytical reaction recipe.

2500 cells/90 μL were cultured by passage in a 96 well cell culture plate, and after 24 hours, the compound represented by Formula 1 was treated with 0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10 (μM). After the reaction for 72 hours, the plate treated with the compound was allowed to stand at room temperature for 30 minutes, and then 100 μL of the reagent was further treated and shaking at room temperature for 10 minutes. Finally, the ratio of the fluorescence value at 570 nm is calculated using an equipment, and the quantitative measurement is performed, and the ability of the compound to inhibit cell growth is confirmed. The measured values measured at the concentrations of the 8 compounds were analyzed using the Prism program (version 5.01, Graphpad Software, Inc.), and the IC ₅₀ value, an indicator of the cell growth inhibitory ability of the compound, was calculated.

	Ba/F3 EGFR wt. IC 50 (μM)	Ba/F3 EGFR del19 IC 50 (μM)	Ba/F3 EGFR L858R IC 50 (μM)
Osimertinib	0.92	0.002	0.007
Example 1	1.8	0.001	0.002
Example 2	0.5	0.009	0.053
Example 3	0.49	0.004	0.028
Example 4	0.41	0.005	0.002
Example 5	0.23	0.008	0.017
Example 6	0.024	0.001	0.003
Example 7	0.4	0.009	0.034
Example 8	0.27	0.009	0.028
Example 9	0.012	0.001	0.001

	Ba/F3 EGFR del19/T790M IC 50 (μM)	Ba/F3 EGFR L858R/T790M IC 50 (μM)	Ba/F3 EGFR del19/T790M/C797S IC 50 (μM)	Ba/F3 EGFR L858R/T790M/C797S IC 50 (μM)
Osimertinib	0.003	0.002	0.33	0.81
Example 1	0.003	0.008	0.12	0.12
Example 2	0.015	0.035	0.24	0.39
Example 3	0.005	0.012	0.16	0.22
Example 4	0.025	0.017	0.32	0.47
Example 5	0.001	0.021	0.092	0.72
Example 6	0.002	0.008	One	0.99
Example 7	0.006	0.01	0.99	0.1
Example 8	0.028	0.013	0.01	0.84
Example 9	0.001	0.001	0.96	0.9

As shown in Table 3 and Table 4 above,

All the compounds of the present invention exhibited relatively weak inhibitory effect against wild-type EGFR in Ba/F3 cell lines, and selectively against EGFR mutations including triple mutations EGFR del19/T790M/C797S, EGFR L858R/T790M/C797S. It showed high cell growth inhibitory ability, and was found to be particularly superior to the case of osimertinib, an existing EGFR TKI drug.

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

<Experimental Example 3> Ba/F3 EGFR exon20 insertion mutant cell growth inhibitory ability in Ba/F3 cell lines

In order to confirm the ability of the compound represented by Example 1 according to the present invention to inhibit cell growth against the Ba/F3 EGFR exon20 insertion mutation in the Ba/F3 cell line, the following experiment was performed. The results are shown in Table 5 below.

The activity measurement of the mutant Ba/F3 EGFR cell line for the compound of the present invention was tested as follows using the CellTiter-Glo system sold by Promega. CellTiter-Glo assay is a method to check cell viability by measuring ATP present in cells in the cell culture state. Ba/F3 EGFR D770delinsGY, H773_V774insH, Y764_V765insHH, V769_D770insASV, D770_N771insSVD mutant cell lines were purchased and used in Pasi A Janne laboratory of Dana-Farber Cancer Institute. Ba/F3 D770delinsGY, H773_V774insH, Y764_V765insHH, V769_D770insASV, D770_N771insSVD mutant cell lines were cultured in an incubator at _{37° C., 5% CO 2} by adding 1 μg/ml of puromycine to RPMI containing 10% FBS and 1% penicillin-streptomycin.

The analysis of the cell growth inhibitory effect of the compound's BaF3 cells overexpressing exon20 insertion mutations was performed according to the following analytical reaction recipe.

1500 cells/100 μL were cultured by passage to a 96 well cell culture plate, and after 24 hours, the compound represented by Example 1 was treated with 0, 30, 100, 300, 1000 (nM). After the reaction for 72 hours, the plate treated with the compound was left at room temperature for 30 minutes, and then 100 μL of CellTiter Glo 2.0 reagent was further treated and shaking at room temperature for 10 minutes. Finally, the luminescence value was measured using a luminometer to confirm the ability of the compound to inhibit cell growth compared to the vehicle (control). The measured values measured at the concentrations of the five compounds were analyzed using the Prism program (version 5.01, Graphpad Software, Inc.), and the IC ₅₀ value, an indicator of the cell growth inhibitory ability of the compound, was calculated.

	Ba/F3 EGFR V769_D770insASV IC 50 (μM)	Ba/F3 EGFR D770delinsGY IC 50 (μM)	Ba/F3 EGFR H773_V774insH IC 50 (μM)	Ba/F3 EGFR Y764_V765insHH IC 50 (μM)	Ba/F3 EGFR D770_N771insSVD IC 50 (μM)
Osimertinib	0.124	0.159	0.191	0.136	0.149
Example 1	0.026	0.021	0.06	0.029	0.027

As shown in Table 5 above,

The compound represented by Example 1 according to the present invention exhibits high cell growth inhibitory ability against Ba/F3 EGFR exon20 insertion mutations in Ba/F3 cell lines, which is the case of osimertinib, an existing EGFR tyrosine kinase inhibitor (TKI) drug. It was confirmed that it is more excellent.

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

<Formulation Example 1> Preparation of powder

2g of derivatives represented by Formula 1

1g lactose

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<Formulation Example 2> Preparation of tablets

100 mg of derivatives represented by Formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet preparation method.

<Formulation Example 3> Preparation of capsules

100 mg of derivatives represented by Formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, a gelatin capsule was filled according to a conventional capsule preparation method to prepare a capsule.

<Formulation Example 4> Preparation of injection

100 mg of derivatives represented by Formula 1

Mannitol 180 mg

Na ₂ HPO ₄ ㆍ2H ₂ O 26 mg

Distilled water 2974 mg

According to a conventional method for preparing injections, injections were prepared by containing the above ingredients in the indicated amount.

<Formulation Example 5> Preparation of health food

500 ng of derivatives represented by Formula 1

The right amount of vitamin mixture

70mg vitamin A acetate

1.0mg vitamin E

0.13mg vitamin

0.15mg vitamin B2

0.5mg vitamin B6

0.2mg vitamin B12

10mg vitamin C

10mg biotin

1.7 mg of nicotinic acid amide

50mg folic acid

0.5mg calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82mg

25.3mg magnesium carbonate

15 mg of potassium monophosphate

Dicalcium Phosphate 55mg

90mg potassium citrate

100mg calcium carbonate

Magnesium chloride 24.8mg

As for the composition ratio of the vitamin and mineral mixture, the ingredients suitable for health food are mixed and formulated as a preferred formulation example, but the mixing ratio may be arbitrarily modified. , To prepare granules, and can be used for preparing a health food composition according to a conventional method.

<Formulation Example 6> Preparation of health drink

500 ng of derivatives represented by Formula 1

1000mg citric acid

100g oligosaccharide

2g plum concentrate

1 g taurine

Total 900ml with purified water

After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized container, sealed and sterilized, and then stored in a refrigerator. It was used in preparation.

The composition ratio is a mixture of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

Claims (10)

1. A compound represented by the following Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof:

   [Formula 1]

   

   In Formula 1,

   R is -OR ^a or -NR ^b1 R ^b2 ,

   R ^a is -(CH ₂ ) _m -NR ^b1 R ^b2 , where m is 1 to 3,

   R ^b1 and R ^b2 are each independently hydrogen or unsubstituted or straight or branched chain C _1-6 ^{alkyl substituted with NR c1} R ^c2 , or R ^b1 and R ^b2 are N, O together with the nitrogen to which they are attached. _{And to form an unsubstituted or straight or branched chain C 1-6} alkyl containing one or more heteroatoms selected from the group consisting of S or a 5 to 7 membered heterocycloalkyl substituted with ^{NR c1} R ^c2,

   R ^c1 and R ^c2 are each independently hydrogen or straight or branched chain C _1-6 alkyl, or, R ^c1 and R ^c2 are selected from the group consisting of N, O and S together with the nitrogen to which they are attached. 5 to 7 membered heterocycloalkyl substituted with unsubstituted or straight or branched C _{1-6 alkyl containing one or more atoms.}
2. The method of claim 1,

   R is

   

   ,

   

   ,

   

   ,

   

   ,

   

   ,

   

   ,

   

   ,

   

   or

   

   Phosphorus compounds, isomers thereof, or pharmaceutically acceptable salts thereof.
3. The method of claim 1,

   The compound represented by Formula 1 is any one selected from the following compound group, an isomer thereof, or a pharmaceutically acceptable salt thereof:

   <1> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-((2-(dimethylamino) Ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide;

   <2> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(4-(dimethylamino)pi Peridin-1-yl)-4-methoxyphenyl)acrylamide;

   <3> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(4- (4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)acrylamide;

   <4> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-morpholino Phenyl)acrylamide;

   <5> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(4- Methylpiperazin-1-yl)phenyl)acrylamide;

   <6> to N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-(2-(dimethylamino) Oxy)-4-methoxyphenyl)acrylamide;

   <7> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(methyl( 2-morpholinoethyl)amino)phenyl)acrylamide;

   <8> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(methyl( 2-(4-methylpiperazin-1-yl)ethyl)amino)phenyl)acrylamide; And

   <9> N-(5-((5-chloro-4-((2-(methylsulfonamido)phenyl)amido)pyrimidin-2-yl)amino)-4-methoxy-2-(2 -Morpholinoethoxy)phenyl)acrylamide.
4. As shown in Scheme 1 below,

   A method for preparing a compound represented by Formula 1 of claim 1 comprising the step of preparing a compound represented by Formula 1 by reacting a compound represented by Formula 2 with a compound represented by Formula 3:

   [Scheme 1]

   

   In Reaction Scheme 1, R is as defined in Formula 1 of claim 1.
5. A pharmaceutical composition for preventing or treating cancer containing the compound represented by Formula 1 of claim 1 as an active ingredient.
6. The method of claim 5,

   The pharmaceutical composition that the compound inhibits EGFR (epidermal growth factor receptor) mutation.
7. The method of claim 6,

   The EGFR mutations are EGFR del19, EGFR L858R, EGFR del19/T790M, EGFR L858R/T790M, EGFR del19/T790M/C797S, EGFR L858R/T790M/C797S, EGFR D770delinsGY, EGFR D770AS H773_V774insVinsH, EGFR del19/T790M, EGFR D770ASH773_V774insH, EGFR D770ASH773, and EGFRVinsD770AS_N, EGFRVinsD770AS Pharmaceutical composition, characterized in that at least one selected from the group.
8. The method of claim 5,

   The cancer is pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, cleft lip cancer, mycosis fungoides, acute myelogenous leukemia, acute lymphocytic leukemia, basal cell cancer, Ovarian epithelial cancer, ovarian germ cell carcinoma, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, barter bulge cancer, bladder cancer, peritoneal cancer , Parathyroid cancer, adrenal cancer, non-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 cancer, kidney cancer, 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, primary site unknown cancer, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal interstitial cancer, Wilm Cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational chorionic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, auditory schwannoma , 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, hematologic cancer and thymic cancer Pharmaceutical composition, characterized in that at least one selected from the group consisting of.
9. Health functional food for preventing or improving cancer containing the compound represented by the formula 1 of claim 1 as an active ingredient.
10. A pharmaceutical composition for preventing or treating diseases related to EGFR mutations containing the compound represented by Formula 1 of claim 1 as an active ingredient.

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