KR102663365B1 - Composition for preventing or treating cancer comprising novel trifluoromethylphenylpyrazol derivative - Google Patents

Abstract

The present invention relates to a composition for preventing or treating cancer containing a novel trifluoromethylphenylpyrazole derivative as an active ingredient, and more specifically, N,N-dimethyl-N'-(3-(1-(4-( Trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H- It relates to a composition for preventing, improving or treating cancer, and a composition for radiotherapy sensitizing agent for cancer, containing novel sulfonamide derivatives including [pyrazol-4-yl)phenyl)azanesulfonamide] as active ingredients. The new sulfonamide derivatives according to the present invention can be used as effective anticancer agents due to their excellent apoptosis effect on cancer cells, and can be used as radiation sensitizer compositions in cancer treatment by lowering the radiation resistance of cancer cells.

Description

Composition for preventing or treating cancer comprising novel trifluoromethylphenylpyrazol derivative as an active ingredient {Composition for preventing or treating cancer comprising novel trifluoromethylphenylpyrazol derivative}

The present invention relates to a composition for preventing or treating cancer containing a novel trifluoromethylphenylpyrazole derivative as an active ingredient, and more specifically, N,N-dimethyl-N'-(3-(1-(4-( Trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H- It relates to a composition for preventing, improving or treating cancer, and a composition for radiotherapy sensitizing agent for cancer containing novel trifluoromethylphenylpyrazole derivatives including [pyrazol-4-yl)phenyl)azanesulfonamide] as active ingredients.

The effective and traditional treatment method for diseases caused by lack of normal control of genes, commonly known as cancer, has been to surgically excise and remove the tumor. However, primary cancer can metastasize to other organs. In cases where surgery is not possible, anticancer drug treatment is widely used. Anticancer drugs used for drug treatment are mainly synthesized from single-molecule substances using organic or inorganic methods. This type of traditional drug treatment involves many side effects, one of which is that the substance used as a drug is an artificially synthesized exogenous substance and the point of action of the anticancer substance is targeted at an already overexpressed protein.

‘Cancer’ refers to a group of diseases that have the characteristics of excessive cell proliferation and infiltration into surrounding tissues when the normal cell death balance is broken. Among these, breast cancer in particular is a cancer with an incidence rate of 25.2% among female cancers worldwide. Between 2008 and 2012, the breast cancer incidence rate continued to rise, increasing by about 20%. In Korea, the incidence rate is 52.1%, making it one of the countries with the highest incidence of breast cancer among OECD countries. It is widely known that breast cancer treatment methods vary depending on the presence or absence of receptors that respond to hormones. To treat breast cancer, surgery, chemotherapy, radiation therapy, and hormone therapy are combined depending on the progress of the cancer.

To date, many natural products, protein or peptide anticancer agents, and chemically synthesized anticancer agents have been developed and used, but most of them not only have serious side effects that affect normal cells in vivo, but also have the same therapeutic effect depending on the cancer type or patient even for the same cancer type. It is common for it not to appear.

In particular, as a method to treat cancer, radiation therapy is an essential treatment method for various cancers, but problems such as acquisition of radiation resistance by cancer cells and damage to normal tissue during high-dose radiation therapy continue to reduce the efficiency of radiation therapy. It is pointed out that Accordingly, the need for substances that can increase the effectiveness of radiation therapy in methods for treating cancer cells is increasing.

Therefore, as a new concept anticancer agent that solves the above problems worldwide, it can selectively remove only cancer cells without affecting normal cells in the body, and can further increase the sensitivity of cancer cells to radiation. The need for a lot of research is required to develop .

Korean Patent No. 10-1855382 (registered on April 30, 2018)

The purpose of the present invention is to provide a novel trifluoromethylphenylpyrazole derivative compound, an isomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer and a health functional food for preventing or improving cancer containing a novel trifluoromethylphenylpyrazole derivative compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. The purpose is to provide a composition.

In addition, another object of the present invention is to provide a pharmaceutical composition for radiotherapy sensitization for cancer containing a novel trifluoromethylphenylpyrazole derivative compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. .

In order to achieve the above object, the present invention provides a compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof.

A compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ₁ is selected from hydrogen, (C1~C6)alkyl, (C1~C6)alkene, (C1~C6)alkyne, substituted or unsubstituted aryl(C1~C6)alkyl, or substituted or unsubstituted aryl, wherein The aryl may be substituted with 1 to 3 substituents and is independently selected from the group consisting of halo, nitro, cyano, hydroxy and oxo, selected from the group consisting of fluoro, chloro and bromo,

R ₂ is selected from hydrogen, (C1~C6)alkyl, (C1~C6)alkylamino or di(C1~C6)alkylamino,

A is any of aryl, pyridine, thiophene, or biphenyl,

_{and ​ ​}

Additionally, the present invention provides a compound represented by the following formula (2), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Formula 2,

Y is hydrogen, CF ₃ , CHF ₂ , CH ₂ F, (C1~C6)alkyl, (C1~C6)alkoxy, or halo selected from the group consisting of fluoro, chloro, and bromo,

n is an integer from 0 to 5,

R is or and R ₁ and R ₂ are each independently selected from hydrogen or substituted or unsubstituted (C1~C6)alkyl, where the alkyl may be substituted with 1 to 3 substituents, and are independently selected from fluoro, chloro and broro. selected from halo or hydroxy selected from the group consisting of

Additionally, the present invention provides a pharmaceutical composition for preventing or treating cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Additionally, the present invention provides a pharmaceutical composition for radiotherapy sensitization for cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention relates to a composition for preventing or treating cancer containing a novel trifluoromethylphenylpyrazole derivative as an active ingredient, and more specifically, N,N-dimethyl-N'-(3-(1-(4-( Trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H- It relates to a composition for preventing, improving or treating cancer, and a composition for radiotherapy sensitizing agent for cancer, containing novel sulfonamide derivatives including [pyrazol-4-yl)phenyl)azanesulfonamide] as active ingredients. The new sulfonamide derivatives according to the present invention can be used as effective anticancer agents due to their excellent apoptosis effect on cancer cells, and can be used as radiation sensitizer compositions in cancer treatment by lowering the radiation resistance of cancer cells.

Figures 1a and 1b show the structural formulas of the compounds used in the present invention.
Figure 2a shows the results of confirming the cancer cell toxicity of various compounds in the MCF7 breast cancer cell line. A was treated with 10 μM, and B was treated with 20 μM.
Figure 2b shows the cancer cytotoxic activity of the compound in various cancer cell lines, including colon cancer (DLD1, HCT116), lung cancer (H1299, A545), and brain cancer (U373, LN18).
Figure 3A shows N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl), named KRCT-1 in the present invention. Azanesulfonamide [N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide;#14-3] shows the chemical structural formula. Figure 3B shows the cancer cell growth inhibition rate upon treatment with KRCT-1 in various breast cancer cell lines, T47D, BT-549, MDA-MB 453, MCF7, and MDA-MB 231, gastric cancer cell line (SNU668), and blood cancer cell line (MV4-11). indicates. Figure 3C shows the results of cancer cell line colony formation analysis upon treatment with KRCT-1 in various breast cancer cell lines: T47D, BT-549, MCF7, and MDA-MB 231. Figure 3D shows the results of tumor sphere formation analysis upon KRCT-1 treatment in various breast cancer cell lines: T47D, BT-549, MCF7, and MDA-MB 231.
Figure 4a shows the results of monitoring the tumor volume and mouse body weight upon administration of KRCT-1 to an immunodeficient mouse tumor model (xenograft mouse model) transplanted with the BT-549 cell line.
Figure 4b shows the tumor growth inhibition effect by KRCT-1 in an immunodeficient mouse tumor model transplanted with the BT-549 cell line. This is the result of confirming the expression of Ki-67 protein, a biomarker that confirms the tumor's proliferation ability, and the tumor dissected from the mouse shown in Figure 4a.
The results of FACS analysis of the apoptosis inducing effect caused by KRCT-1 treatment in the breast cancer cell line shown in Figure 5a are shown by Annexin V/PI staining.
Figure 5b shows the results of Western blot confirmation of the apoptosis effect caused by KRCT-1 treatment in each breast cancer cell line, BT549 (A), T47D (B), and MCF7 (C).
Figure 6a shows the results of confirming the formation of cancer cell colonies in breast cancer cell lines BT549, T47D, and MCF7 to confirm the radiation sensitivity of KRCT-1.
Figure 6b shows the results of confirming DNA damage in breast cancer cell lines BT549, T47D, and MCF7 to confirm the radiation sensitivity of KRCT-1. DNA damage markers were confirmed through Western blot and immunocytochemistry (ICC).
Figure 7a shows the results of 104 kinase profiling to explore the molecular biological mechanism of action of KRCT-1.
Figure 7b shows the in vitro inhibition of Trk-A kinase activity by KRCT-1.
Figure 7c shows the results confirming that downstream signaling of Trk-A kinase in cancer cells is inhibited by KRCT-1.

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 ₁ is selected from hydrogen, (C1~C6)alkyl, (C1~C6)alkene, (C1~C6)alkyne, substituted or unsubstituted aryl(C1~C6)alkyl, or substituted or unsubstituted aryl, wherein The aryl may be substituted with 1 to 3 substituents and is independently selected from the group consisting of halo, nitro, cyano, hydroxy and oxo, selected from the group consisting of fluoro, chloro and bromo,

R ₂ is selected from hydrogen, (C1~C6)alkyl, (C1~C6)alkylamino or di(C1~C6)alkylamino,

A is any of aryl, pyridine, thiophene, or biphenyl,

_{and ​ ​}

Preferably, the compound represented by Formula 1 is N-methyl-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide [N-methyl-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol -4-yl)phenyl)ethanesulfonamide; #14-1], N-(4-chlorobenzyl)-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide [ N-(4-chlorobenzyl)-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide; #14-2], N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide [N ,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-3], N,N-dimethyl-N'-methyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ah Zansulfonamide [N,N-dimethyl-N'-methyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-4], N,N-dimethyl-N'-(4-chlorobenzyl)-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazole-4- 1) phenyl) azanesulfonamide [N,N-dimethyl-N'-(4-chlorobenzyl)-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl) phenyl)azanesulfonamide; #14-5], N,N-dimethyl-N'-(3-(1-(4-phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N '-(3-(1-(4-phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-11], N,N-dimethyl-N'-(3-(1-(4-(methyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N- dimethyl-N'-(3-(1-(4-(methyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-12], N,N-dimethyl-N'-(3-(1-(4-(methoxy)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N -dimethyl-N'-(3-(1-(4-(methoxy)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-13], N,N-dimethyl-N'-(3-(1-(pyridin-4-yl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl -N'-(3-(1-(pyridin-4-yl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-14], N,N-dimethyl-N'-(3-(1-(thiophen-4-yl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N- dimethyl-N'-(3-(1-(thiophen-4-yl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-15], N,N-dimethyl-N'-(3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl -N'-(3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-16], N,N-dimethyl-N'-(3-(1-(4-([1,1'-biphenyl]-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-(3-(1-(4-([1,1'-biphenyl]-1H-pyrazol-4-yl)phenyl)azanesulfonamide; #14-17], N,N -Dimethyl-N'-isobutyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N- dimethyl-N'-isobutyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide #14-18], N,N-dimethyl-N '-allyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N-dimethyl-N'- allyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide #14-19], N,N-dimethyl-N'-phenyl-N; '-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N-dimethyl-N'-phenyl-N'- (3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide #14-20] or N-(3-(1-(4-(trifluoromethyl)) Phenyl)-1H-pyrazol-4-yl)phenyl)piperidine-1-sulfonamide [N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl) piperidine-1-sulfonamide; #14-21], but is not limited thereto.

The chemical structures of the compounds #14-1 to #14-5 and #14-11 to #14-21 are as follows.

Additionally, the present invention provides a compound represented by the following formula (2), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Formula 2,

Y is hydrogen, CF ₃ , CHF ₂ , CH ₂ F, (C1~C6)alkyl, (C1~C6)alkoxy, or halo selected from the group consisting of fluoro, chloro, and bromo,

n is an integer from 0 to 5,

R is or and R ₁ and R ₂ are each independently selected from hydrogen or substituted or unsubstituted (C1~C6)alkyl, where the alkyl may be substituted with 1 to 3 substituents, and are independently selected from fluoro, chloro and broro. selected from halo or hydroxy selected from the group consisting of

Preferably, the compound represented by Formula 2 is (E)-N-ethyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl ) Methanimine oxide [(E)-N-ethyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide; #14-6], (E)-N-isopropyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide [ (E)-N-isopropyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide; #14-7], (E)-N-tert-butyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide [(E)-N-tert-butyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide; #14-8], (E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-ethyl oxime [(E)-3-(1 -(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-ethyl oxime; #14-9] or (E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-(tert-butyl)oxime[(E)- 3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-(tert-butyl) oxime; #14-10], but is not limited thereto.

The chemical structures of compounds #14-6 to #14-10 are as follows.

The pharmaceutically acceptable salts include hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, and fumaric acid. It may be selected from the group consisting of salts, methanesulfonate, benzenesulfonate, paratoluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt, but is not limited thereto.

Additionally, the present invention provides a pharmaceutical composition for preventing or treating cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

It was confirmed that the pharmaceutical composition of the present invention reduces the expression of survivin, which is essential for the survival of cancer cells, and induces cell death. Survivin has a high expression rate in various cancers, including lung cancer, breast cancer, prostate cancer, and ovarian cancer, and is involved in the survival and proliferation of cancer cells as a protein that inhibits cell death. The survivin is being studied extensively as a diagnostic marker and target for developing targeted anticancer drugs. Accordingly, the present inventors confirmed that treating cancer cells with KRCT-1, which inhibits the expression of survivin, can effectively inhibit cancer cell proliferation.

When the composition of the present invention is a pharmaceutical composition, it can be prepared using pharmaceutically suitable and physiologically acceptable auxiliaries in addition to the active ingredients, and the auxiliaries include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, Solubilizers such as lubricants, glidants, or flavoring agents can be used. For administration, the pharmaceutical composition of the present invention can be preferably formulated as a pharmaceutical composition containing one or more pharmaceutically acceptable carriers in addition to the active ingredient. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these ingredients can be mixed and used, and other common additives such as antioxidants, buffers, and bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

The pharmaceutical preparation form of the pharmaceutical composition of the present invention may be granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions, and sustained-release preparations of the active compound. You can. The pharmaceutical composition of the present invention can be administered in any conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal routes. It can be administered. The effective amount of the active ingredient of the pharmaceutical composition of the present invention refers to the amount required for the prevention or treatment of disease. Therefore, the type of disease, the severity of the disease, the type and content of the active ingredient and other ingredients contained in the composition, the type of dosage form and the patient's age, weight, general health condition, gender and diet, administration time, administration route and composition. It can be adjusted depending on a variety of factors, including secretion rate, duration of treatment, and concurrent medications.

In addition, the pharmaceutical composition according to the present invention may contain a trifluoromethylphenylpyrazole derivative compound as an active ingredient at a concentration of 5 to 150 μM, preferably at a concentration of 10 to 120 μM, and more preferably at a concentration of 30 to 150 μM. It may be included at an effective concentration of 100 μM.

'Cancer' in the present invention refers to a group of diseases characterized by excessive cell proliferation and infiltration into surrounding tissues when the normal cell death balance is broken. The cancer of the present invention includes brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, nasal cavity/paranasal sinus cancer, and nasopharyngeal cancer. , salivary gland cancer, hypopharyngeal cancer, thyroid cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, stomach cancer, liver cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer, small intestine cancer, Colon cancer, anal cancer, bladder cancer, kidney cancer, male genital cancer, penile cancer, urethral cancer, prostate cancer, female genital cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female external genital cancer, female urethra It may be selected from the group consisting of cancer, skin cancer, myeloma, leukemia, and malignant lymphoma, preferably prostate cancer including breast cancer, lung cancer, and stomach cancer, and most preferably breast cancer, but is not limited thereto.

The pharmaceutical composition of the present invention can be used in combination with radiation therapy. In particular, the pharmaceutical composition of the present invention can increase the sensitivity to radiation of cancer cells or cancer cells with radioresistance, so it can be used in combination with radiation therapy. When treating cancer cells, a feature exists in that it can increase the effect of radiation therapy.

In addition, the present invention provides a health functional food composition for preventing or improving cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the term "health functional food" refers to a food manufactured and processed using specific ingredients as raw materials or specific ingredients contained in food raw materials by methods such as extraction, concentration, purification, mixing, etc. for the purpose of health supplementation. It refers to a food designed and processed so that its ingredients can fully exert bioregulatory functions on the living body, such as biological defense, regulation of biological rhythm, prevention and recovery of disease, etc., and can perform functions related to disease prevention or health recovery. Say what you can.

When the trifluoromethylphenylpyrazole derivative compound according to the present invention is used as a health functional food, it can be added as is or used together with other foods or food ingredients, and can be selected and used appropriately as needed.

Additionally, there is no particular limitation on the type of health functional food in which the trifluoromethylphenylpyrazole derivative compound according to the present invention can be used. For example, ramen, other noodles, beverages, tea, drinks, alcoholic beverages, various soups, meat, sausages, bread, chocolate, candy, confectionery, pizza, gum, dairy products including ice cream, or vitamin complexes. In addition, the health functional food according to the present invention can be mixed with other appropriate auxiliary ingredients and known additives that can be commonly contained in health functional foods according to the selection of a person skilled in the art in addition to the trifluoromethylphenylpyrazole derivative compound.

Additionally, the present invention provides a pharmaceutical composition for radiotherapy sensitization for cancer containing the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Treating cancer cells with the trifluoromethylphenylpyrazole derivative compound of the present invention increases the sensitivity of cancer cells to radiation, thereby enhancing the death of cancer cells.

The criteria for radiation sensitivity or radiation resistance can be classified by the slope value of a linear model created using cell survival rate when irradiated with 0 to 3 Gy of gamma radiation. This classification method was reported by Jerry R. Williams et al. in a paper (Acta Oncologica, 46, 628-638, 2007). As used in the present invention, the term "radiation sensitivity" refers to cells having characteristics such that the slope value falls within the range of 0.00 to 0.30, and "radiation resistance" refers to cells having characteristics falling within the range of 0.31 to 1.00. .

In the present invention, the cancer includes brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brain stem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, and nasal/paranasal sinus cancer. , nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, stomach cancer, liver cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer, Small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, male genital cancer, penile cancer, urethral cancer, prostate cancer, female genital cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, and female external genital cancer. , may be selected from the group consisting of female urethral cancer, skin cancer, myeloma, leukemia, and malignant lymphoma, preferably prostate cancer including breast cancer, lung cancer, or stomach cancer, and most preferably breast cancer, but is limited thereto. That is not the case.

The pharmaceutical composition for radiotherapy sensitizing agent for cancer according to the present invention can be used in combination with radiotherapy.

Radiation therapy used in the present invention is a local treatment method that damages the DNA of malignant cells. Normal cells have a greater ability to repair this damage than tumor cells, and radiotherapy is known to take advantage of this difference.

The radiotherapy may include external beam radiation (x-rays, γ-rays, protons and neutrons), brachytherapy and implantation of radioactive substances, including 2-D, 3-D, confocal, intensity-modulated ( It can be administered by IMRT) and image-guided (IGRT) approaches. The standard radiation therapy that can be used in the cancer treatment of the present invention may be a method of providing 2.5 Gy/day with a total dose of about 60 Gys (50 to 70 Gys). However, one skilled in the art can select the desired radiation dose based on the particular subject, device, and tumor type.

The pharmaceutical composition for radiotherapy sensitizing agent for cancer according to the present invention constitutes an improvement over existing radiotherapy treatments by increasing the sensitivity of cancer cells to radiation. For example, use of the pharmaceutical composition for radiotherapy sensitizing agent for cancer of the present invention causes cancer cells to respond to radiation by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or It has a sensitivity higher than this. As used herein, the terms “sensitivity” and “radiation sensitivity” refer to the number of viable cells based on radiation dose. Therefore, an increase in radiation sensitivity may mean a decrease in the number of cells surviving a given radiation dose, a decrease in the radiation dose required for lethality, or a combination thereof.

Since the pharmaceutical composition for radiotherapy sensitizing agent for cancer of the present invention can increase the sensitivity of cancer cells not only to radiotherapy but also to chemotherapy, the radiosensitizing agent of the present invention is used as a sensitizing agent to increase sensitivity in chemotherapy. Cases where it is used may also fall within the scope of the present invention.

In the present invention, the term “administration in combination” refers to administering radiation together in an anti-cancer process to treat various types of cancer cells. Specifically, radiation therapy may be combined in an anticancer process to treat cancer cells such as solid cancers such as lung cancer, breast cancer, colon cancer, ovarian cancer, head and neck cancer, and brain cancer.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

< Manufacturing example 1> 3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4- yl )aniline

Step 1 : 4-iodobenzotrifluoride (1.00 g, 3.68 mmol), 4-chloro-1H-pyrazole (415 mg, 4.05 mmol), cesium carbonate (2.40 g, 7.36 mmol), and CuI (105 mg, 0.55 mmol) as reaction solvent. Phosphorus was added to anhydrous toluene (35 mL) and stirred at 110°C for 24 hours. After confirming the reaction product by TLC, the reaction solution was diluted with EtOAc and washed with purified water. The organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by silica column chromatography to obtain 1.12 g of the title compound (Compound 1). ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (s, 1H), 8.02 (s, 1H), 7.89 (2H, d, J=8.77 Hz), 7.76 (2H, d, J=8.77 Hz) ; LC-MS (ESI, m/z) = 247.0 (M+H ⁺ ).

Step 2 : Compound 1 (800 mg, 3.25 mmol) obtained from Step 1 was placed in a reaction vessel, and 3-aminophenylboronic acid (534 mg, 3.90 mmol), Pd(OAc) ₂ (73 mg, 0.32 mmol), and sSPhos ( 250 mg, 0.49 mmol) and Na ₂ CO ₃ (689 mg, 6.50 mmol) were added to anhydrous dioxane (25 mL) and then treated with N ₂ (g) for 5 minutes. The reaction was stirred at 110°C for 12 hours, the product was confirmed by TLC, and the solvent, dioxane, was removed by concentration under reduced pressure. The remaining reaction was diluted with EtOAc and washed with purified water. The organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by silica column chromatography to obtain 921 mg of the title compound (Compound 2). ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.21 (s, 1H), 8.08 (s, 1H), 7.81 (2H, d, J=8.74 Hz), 7.67 (2H, d, J=8.77 Hz) , 7.45 (1H, m), 7.02 (1H, d, J=7.82 Hz), 6.89 (1H, s), 6.76 (1H, d, J=7.69 Hz), 4.66 (2H, s); LC-MS (ESI, m/z) = 304.1 (M+H ⁺ ).

< Manufacturing example 2> 2-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)benzaldehyde

Compound 1 (800 mg, 3.25 mmol) obtained from Step 1 of Preparation Example 1 was placed in a reaction vessel, and (3-formylphenyl)boronic acid (585 mg, 3.90 mmol) and Pd(PPh ₃ ) ₄ (375 mg, 0.33 mmol) were added to the reaction vessel. ), Na ₂ CO ₃ (689 mg, 6.54 mmol) was added to dioxane (32 mL) and then treated with N ₂ (g) for 5 minutes. The reaction was stirred at 90°C for 8 hours, the product was confirmed by TLC, and the solvent, dioxane, was removed by concentration under reduced pressure. The reaction concentrate was diluted with EtOAc and washed with purified water. The organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by silica column chromatography to obtain 870 mg of the title compound (Compound 3). ¹ H-NMR (300 MHz, CDCl ₃ ): δ 9.85 (s, 1H), 8.61 (s, 1H), 8.21 (s, 1H), 8.08 (s, 1H), 7.95 (1H, d, J=7.86 Hz), 7.81 (2H, d, J=8.74 Hz), 7.67 (2H, d, J=8.77 Hz), 7.45 (1H, m), 7.22 (1H, d, J=7.82 Hz); LC-MS (ESI, m/z) = 317.1 (M+H ⁺ ).

< Example 1> Compound synthesis

1.#14- 1 and #14-2 Synthesis scheme

#14-1: N-methyl-N-(3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)phenyl)ethanesulfonamide

Step 1: Compound 2, 3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)aniline (591 mg, 1.95 mmol) synthesized in Preparation Example 1 was added to a reaction flask and 20 mL of Dissolve in anhydrous DCM solvent. Ethanesulfonyl chloride (204 μL, 2.16 mmol) and pyridine (174 μL, 2.16 mmol) were added and stirred at room temperature for 12 hours. The reaction product was confirmed by TLC, then diluted with DCM and washed with NH ₄ Cl aqueous solution. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 603 mg of Compound 4. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 10.51 (s, 1H), 8.20 (s, 1H), 8.04 (s, 1H), 7.83 (d, 2H, J =8.77 Hz), 7.73 (d, 2H, J = 8.77 Hz), 7.62 (s, 1H), 7.51 (d, 1H, J = 7.44 Hz), 7.42 (t, 1H, J = 7.71 Hz), 7.27 (d, 1H, J = 7.71 Hz) , 3.12 (q, 2H, J = 7.44 Hz), 1.42 (t, 3H, J = 7.71 Hz); LC-MS (ESI, m/z) = 396.1 (M+H ⁺ ).

Step 2 : Compound 4 (256 mg, 0.65 mmol) obtained from Step 1 above was placed in a reaction flask and dissolved in 7 mL of anhydrous MeCN solvent. Methyl iodide (38 μL, 0.65 mmol) and K ₂ CO ₃ (90 mg, 0.65 mmol) were added and stirred at 80°C for 2 hours. The reaction product was confirmed by TLC, diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 220 mg of the title compound #14-1. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (s, 1H), 8.02 (s, 1H), 7.89 (d, 2H, J = 8.77 Hz), 7.76 (d, 2H, J = 8.77 Hz) , 7.62 (s, 1H), 7.49 (d, 1H, J = 7.44 Hz), 7.45 (t, 1H, J = 7.71 Hz), 7.30 (d, 1H, J = 7.71 Hz), 3.40 (s, 3H) , 3.12 (q, 2H, J = 7.44 Hz), 1.42 (t, 3H, J = 7.71 Hz); LC-MS (ESI, m/z) = 410.2 (M+H ⁺ ).

#14-2: N-(4- chlorobenzyl )-N-(3-(1-(4-( trifluoromethyl ) phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide

Compound 4 (67 mg, 0.17 mmol) obtained from step 1 above was placed in a reaction flask and dissolved in 3 mL of anhydrous MeCN solvent. 1-(Bromomethyl)-4-chlorobenzene (82 mg, 0.21 mmol) and K ₂ CO ₃ (29 mg, 0.21 mmol) were added and stirred at 80°C for 2 hours. The reaction product was confirmed by TLC, diluted with EtOAc, and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 60 mg of the title compound #14-2. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.12 (s, 1H), 7.94 (s, 1H), 7.87 (d, 2H, J = 8.54 Hz), 7.75 (d, 2H, J = 8.54 Hz) , 7.46 (m, 1H), 7.38 (m, 2H), 7.24 (m, 3H), 7.17 (m, 1H), 4.88 (s, 2H), 3.15 (q, 2H, J = 7.35 Hz), 1.47 ( t, 3H, J = 7.35 Hz); LC-MS (ESI, m/z) = 520.1 (M+H ⁺ ).

2. #14-3, #14-4, #14-5, #14-11 to #14-21 synthesis scheme

#14-3: N,N - dimethyl -N’-(3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)phenyl)azanesulfonamide

Compound 2, 3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)aniline (591 mg, 1.95 mmol) synthesized in Preparation Example 1 was added to a reaction flask and added to 20 mL of anhydrous DCM solvent. Dissolve it with Dimethylsulfamoyl chloride (671 μL, 2.16 mmol) and pyridine (174 μL, 2.16 mmol) were added and stirred at room temperature for 12 hours. The reaction product was confirmed by TLC, then diluted with DCM and washed with NH ₄ Cl solution. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 781 mg of the title compound #14-3. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.21 (S, 1H), 8.01 (s, 1H), 7.89 (d, 2H, J= 8.51 Hz), 7.76 (d, 2H, J= 8.51 Hz) , 7.40 (m, 1H), 7.36 (m, 2H), 7.09 (m, 1H), 6.54 (s, 1H), 2.89 (s, 6H); LC-MS (ESI, m/z) = 411.1 (M+H ⁺ ).

#14-4: N, N - dimethyl -N’-methyl-N’-(3-(1-(4-( trifluoromethyl )phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide

The compound of Example #14-3 (100 mg, 0.24 mmol) was placed in a reaction flask and dissolved in 3 mL of anhydrous MeCN solvent. Iodomethane (17 μL, 0.28 mmol) and K ₂ CO ₃ (67 mg, 0.50 mmol) were added and stirred at 80°C for 2 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 94 mg of the title compound #14-4. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (s, 1H), 8.02 (s, 1H), 7.89 (d, 2H, J = 8.62 Hz), 7.76 (d, 2H, J = 8.62 Hz) , 7.62 (m, 1H), 7.48 (m, 1H), 7.44 (t, 1H, J = 7.63 Hz), 7.32 (m, 1H), 3.33 (s, 3H), 2.83 (s, 6H); LC-MS (ESI, m/z) = 425.1 (M+H ⁺ ).

#14-5: N, N - dimethyl -N’-(4- chlorobenzyl )-N’-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide

The compound of Example #14-3 (100 mg, 0.24 mmol) was placed in a reaction flask and dissolved in 3 mL of anhydrous MeCN solvent. 4-Chlorobenzyl bromide (55 mg, 0.28 mmol) and K ₂ CO ₃ (67 mg, 0.50 mmol) were added and stirred at 80°C for 2 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 106 mg of the title compound #14-5. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.12 (s, 1H), 7.94 (s, 1H), 7.88 (d, 2H, J= 8.83 Hz), 7.75 (d, 2H, J= 8.83 Hz) , 7.46 (m, 1H), 7.37 (t, 1H, J= 7.61 Hz), 7.26 (s, 1H), 7.24 (d, 2H, J= 10.96 Hz), 7.15 (m, 1H), 4.78 (s, 2H), 2.78 (s, 6H); LC-MS (ESI, m/z) = 535.1 (M+H ⁺ ).

#14-11: N,N - dimethyl -N’-(3-(1-(4-phenyl)-1H- pyrazol -4-yl)phenyl)azanesulfonamide

Step 1: React iodobenzene (1.00 g, 4.90 mmol) and 4-chloro-1H-pyrazole (552 mg, 5.39 mmol) in a similar manner to Preparation Example 1 to obtain 750 mg of the title compound 3-(1-phenyl-1H- pyrazol-4-yl)aniline was obtained. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.19 (s, 1H), 8.10 (s, 1H), 7.64 (2H, d, J=8.71 Hz), 7.42 (2H, m), 7.02 (1H, d, J=7.69 Hz), 6.84 (1H, s), 6.78 (1H, d, J=7.74 Hz), 4.67 (2H, s); LC-MS (ESI, m/z) = 236.1 (M+H ⁺ ).

Step 2: A method similar to Example #14-3 using the compound 3-(1-phenyl-1H-pyrazol-4-yl)aniline (200 mg, 0.85 mmol) obtained from Step 1 above as a starting material for the reaction. 87 mg of the title compound was obtained by reaction. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 10.51 (s, 1H), 8.19 (S, 1H), 8.01 (s, 1H), 7.87-7.74 (m, 5H), 7.40 (m, 1H), 7.36 (m, 2H), 7.12 (m, 1H), 6.52 (s, 1H), 2.89 (s, 6H); LC-MS (ESI, m/z) = 344.2 (M+H ⁺ ).

#14-12: N,N - dimethyl -N’-(3-(1-(4-(methyl)phenyl)-1H- pyrazol -4- yl )phenyl) azanesulfonamide

Step 1: Using 1-iodo-4-methylbenzene (1.00 g, 4.58 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 632 mg of the title compound 3-(1-(p-tolyl)-1H- pyrazol-4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.21 (s, 1H), 8.09 (s, 1H), 7.82 (2H, d, J=8.77 Hz), 7.69 (2H, d, J=8.77 Hz), 7.00 (1H, d, J=7.85 Hz), 6.86 (1H, s), 6.76 (1H, d, J=7.66 Hz), 4.69 (s, 2H), 2.43 (s, 3H); LC-MS (ESI, m/z) = 250.08 (M+H ⁺ ).

Step 2: Compound 3-(1-(p-tolyl)-1H-pyrazol-4-yl)aniline (200 mg, 0.83 mmol) obtained from Step 1 was used as a starting material for the reaction, Example #14- By reacting in a similar manner to 3, 58 mg of the title compound was obtained. 1H-NMR (300 MHz, CDCl ₃ ): δ 10.50 (s, 1H), 8.20 (S, 1H), 8.00 (s, 1H), 7.89 (d, 2H, J=8.48 Hz), 7.76 (d, 2H) , J=8.51 Hz), 7.37 (m, 1H), 7.38 (m, 2H), 7.08 (m, 1H), 6.54 (s, 1H), 2.44 (s, 3H), 2.88 (s, 6H); LC-MS (ESI, m/z) = 357.1 (M+H ⁺ ).

#14-13: N,N - dimethyl -N’-(3-(1-(4-( methoxy )phenyl)-1H- pyrazol -4-yl)phenyl)azanesulfonamide

Step 1: Using 1-iodo-4-methoybenzene (1.00 g, 4.27 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 734 mg of the title compound 3-(1-(4-methoxyphenyl)-1H- pyrazol-4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.20 (s, 1H), 8.03 (s, 1H), 7.78 (2H, d, J=8.74 Hz), 7.64 (2H, d, J=8.74 Hz), 6.97 (1H, d, J=7.74 Hz), 6.85 (1H, s), 6.76 (1H, d, J=7.66 Hz), 4.69 (s, 2H), 3.82 (s, 3H); LC-MS (ESI, m/z) = 266.0 (M+H ⁺ ).

Step 2: Compound 3-(1-(4-methoxyphenyl)-1H-pyrazol-4-yl)aniline (200 mg, 0.76 mmol) obtained from Step 1 was used as a starting material for the reaction, Example #14- By reacting in a similar manner to 3, 61 mg of the title compound was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.15 (S, 1H), 8.03 (s, 1H), 7.85 (d, 2H, J=8.45 Hz), 7.69 (d, 2H, J=8.48 Hz), 7.34 (m, 1H), 7.30 (m, 2H), 7.09 (m, 1H), 6.52 (s, 1H), 3.81 (s, 3H), 2.88 (s, 6H); LC-MS (ESI, m/z) = 373.1 (M+H ⁺ ).

#14-14: N, N - dimethyl -N’-(3-(1-( pyridine -4- yl )-1H- pyrazol -4- yl )phenyl) azanesulfonamide

Step 1: Using 4-iodopyridine (1.00 g, 4.87 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 541 mg of the title compound 3-(1-(pyridin-4-yl)-1H-pyrazol- 4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.32 (s, 1H), 8.12 (s, 1H), 7.98 (2H, d, J=8.64 Hz), 7.83 (2H, d, J=6.64 Hz), 7.01 (m, 1H), 6.97 (1H, d, J=7.72 Hz), 6.85 (1H, s), 6.72 (1H, d, J=7.72 Hz), 4.69 (s, 2H); LC-MS (ESI, m/z) = 237.0 (M+H ⁺ ).

Step 2: Example # By reacting in a similar manner to 14-3, 47 mg of the title compound was obtained. 1H-NMR (300 MHz, CDCl3): δ 10.53 (s, 1H), 8.21 (S, 1H), 8.19 (s, 1H), 7.98 (d, 2H, J=8.42 Hz), 7.79 (d, 2H, J=8.42 Hz), 7.31 (m, 1H), 7.29 (m, 2H), 7.09 (m, 1H), 6.51 (s, 1H), 2.87 (s, 6H); LC-MS (ESI, m/z) = 344.1 (M+H ⁺ ).

#14-15: N,N - dimethyl -N’-(3-(1-( thiophen -4- yl )-1H- pyrazol -4-yl)phenyl)azanesulfonamide

Step 1: Using 2-iodothiophen (1.00 g, 4.21 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 708 mg of the title compound 3-(1-(thiophen-2-yl)-1H-pyrazol- 4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.39 (s, 1H), 8.09 (s, 1H), 7.89 (1H, d, J=8.64 Hz), 7.75 (1H, d, J=8.64 Hz), 7.65 (m, 1H), 7.01 (m, 1H), 6.97 (1H, d, J=7.72 Hz), 6.85 (1H, s), 6.72 (1H, d, J=7.72 Hz), 4.67 (s, 2H) ; LC-MS (ESI, m/z) = 242.3 (M+H ⁺ ).

Step 2: Compound 3-(1-(thiophen-2-yl)-1H-pyrazol-4-yl)aniline obtained from Step 1 was used as a starting material for the reaction in a similar manner to Example #14-3. By reaction, 63 mg of the title compound was obtained. 1H-NMR (300 MHz, CDCl3): δ 10.50 (s, 1H), 8.39 (s, 1H), 8.12 (s, 1H), 7.95 (1H, d, J=8.64 Hz), 7.79 (1H, d, J=8.64 Hz), 7.64 (m, 1H), 7.01 (m, 1H), 6.98 (1H, d, J=7.72 Hz), 6.85 (1H, s), 6.71 (1H, d, J=7.72 Hz) , 6.52 (s, 1H), 2.88 (s, 6H); LC-MS (ESI, m/z) = 348.9 (M+H ⁺ ).

#14-16: N,N - dimethyl -N’-(3-(1-(4- fluorophenyl )-1H- pyrazol -4- yl )phenyl) azanesulfonamide

Step 1: Using 1-fluoro-4-iodobenzene (1.00 g, 4.51 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 672 mg of the title compound 3-(1-(4-fluorophenyl)-1H- pyrazol-4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.27 (s, 1H), 8.08 (s, 1H), 7.82 (2H, d, J=8.74 Hz), 7.68 (2H, d, J=8.74 Hz), 7.46 (m, 1H), 7.04 (1H, d, J=7.82 Hz), 6.85 (1H, s), 6.72 (1H, d, J=7.69 Hz), 4.64 (s, 2H); LC-MS (ESI, m/z) = 254.1 (M+H ⁺ ).

Step 2: Compound 3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)aniline (200 mg, 0.78 mmol) obtained from Step 1 was used as a starting material for the reaction, Example #14- By reacting in a similar manner to 3, 65 mg of the title compound was obtained. 1H-NMR (300 MHz, CDCl3): δ 10.50 (s, 1H), 8.26 (s, 1H), 8.08 (s, 1H), 7.84 (2H, d, J=8.71 Hz), 7.69 (2H, d, J=8.74 Hz), 7.48 (m, 1H), 7.06 (1H, d, J=7.85 Hz), 6.85 (1H, s), 6.72 (1H, d, J=7.69 Hz), 6.52 (s, 1H) , 2.90 (s, 6H); LC-MS (ESI, m/z) = 361.1 (M+H ⁺ ).

#14-17: N,N - dimethyl -N’-(3-(1-(4-([1,1’-biphenyl]-1H- pyrazol -4- yl )phenyl) azanesulfonamide

Step 1: Using 1-iodobiphenyl (1.00 g, 3.57 mmol) as a starting material, react in a similar manner to Preparation Example 1 to obtain 812 mg of the title compound 3-(1-([1,1'-biphenyl]-4- yl)-1H-pyrazol-4-yl)aniline was obtained. 1H-NMR (300 MHz, CDCl3): δ 8.32 (s, 1H), 8.14 (s, 1H), 7.81 (2H, d, J=8.77 Hz), 7.72 (2H, d, J=8.77 Hz), 7.69 -7.52 (m, 5H), 7.46 (m, 1H), 7.01 (1H, d, J=7.82 Hz), 6.85 (1H, s), 6.78 (1H, d, J=7.66 Hz), 4.65 (s, 2H); LC-MS (ESI, m/z) = 312.1 (M+H ⁺ ).

Step 2: Compound 3-(1-([1,1'-biphenyl]-4-yl)-1H-pyrazol-4-yl)aniline (200 mg, 0.64 mmol) obtained from Step 1 was used at the start of the reaction. 61 mg of the title compound was obtained by reacting in a similar manner to Example #14-3. 1H-NMR (300 MHz, CDCl3): δ 10.48 (s, 1H), 8.32 (s, 1H), 8.11 (s, 1H), 7.83 (2H, d, J=8.77 Hz), 7.70 (2H, d, J=8.77 Hz), 7.70-7.52 (m, 5H), 7.46 (m, 1H), 7.03 (1H, d, J=7.74 Hz), 6.81 (1H, s), 6.73 (1H, d, J=7.69 Hz), 6.50 (s, 1H), 2.91 (s. 6H); LC-MS (ESI, m/z) = 419.0 (M+H ⁺ ).

#14-18: N,N - dimethyl -N’- isobutyl -N’-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide

1-iodo-2-methylpropan (53 mg, 0.29 mmol) was added to the starting material #14-3 (100 mg, 0.24 mmol) and reacted in a similar manner to Example #14-4 to obtain 32 mg of the title compound. did. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.22 (s, 1H), 8.01 (s, 1H), 7.87 (d, 2H, J = 8.62 Hz), 7.71 (d, 2H, J = 8.62 Hz) , 7.59 (m, 1H), 7.49 (m, 1H), 7.42 (t, 1H, J = 7.60 Hz), 7.30 (m, 1H), 2.98 (d, J=7.72 Hz, 2H), 2.82 (s, 6H), 1.68 (m, 1H), 0.92 (d, 6H, J=6.8 Hz); LC-MS (ESI, m/z) = 466.9 (M+H ⁺ ).

#14-19: N,N - dimethyl -N’- allyl -N’-(3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)phenyl)azanesulfonamide

3-bromo-1-propene (35 mg, 0.29 mmol) was added to the starting material #14-3 (100 mg, 0.24 mmol) and reacted in a similar manner to Example #14-4 to obtain 41 mg of the title compound. did. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (s, 1H), 8.04 (s, 1H), 7.86 (d, 2H, J = 8.45 Hz), 7.72 (d, 2H, J = 8.45 Hz) , 7.61 (m, 1H), 7.52 (m, 1H), 7.41 (t, 1H, J = 7.63 Hz), 7.31 (m, 1H), 5.85 (m, 1H), 5.19 (d, J=17.1 Hz, 1H), 5.17 (d, J=17.1 Hz, 1H), 3.73 (d, J=6.2 Hz, 1H), 2.82 (s, 6H); LC-MS (ESI, m/z) = 450.9 (M+H ⁺ ).

#14-20: N, N - dimethyl -N’-phenyl-N’-(3-(1-(4-( trifluoromethyl )phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide

Starting materials #14-3 (100 mg, 0.24 mmol), phenylboronic acid (36 mg, 0.29 mmol), Tetrakis(acetonitrile)copper(I) hexafluorophosphate (108 mg, 0.29 mmol), N-methylpiperidine (0.17 mL, 1.46 mmol) was added to anhydrous MeCN and stirred at room temperature for 12 hours. The reaction product was confirmed by TLC, then diluted with DCM and washed with NH4Cl solution. The separated organic layer was dehydrated with anhydrous Na2SO4, concentrated under reduced pressure, and separated by column chromatography to obtain 29 mg of the title compound #14-20.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (S, 1H), 8.08 (s, 1H), 7.88 (d, 2H, J= 8.48 Hz), 7.79 (d, 2H, J= 8.48 Hz) , 7.42 (m, 1H), 7.31 (m, 2H), 7.29-7.15 (m, 5H), 7.09 (m, 1H), 2.87 (s, 6H); LC-MS (ESI, m/z) = 486.9 (M+H ⁺ ).

#14-21: N-(3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)phenyl)piperidine-1-sulfonamide

Compound synthesized in Preparation Example 1, 3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)aniline (100 mg, 0.33 mmol) and Piperidine-1-sulfonyl chloride (56 uL, 0.39 uL) mmol) was subjected to a similar reaction as Example #14-3 to obtain 42 mg of the title compound. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.23 (S, 1H), 7.98 (s, 1H), 7.84 (d, 2H, J= 8.31 Hz), 7.74 (d, 2H, J= 8.31 Hz) , 7.36 (m, 1H), 7.33 (m, 2H), 7.01 (m, 1H), 3.07 (t, J= 7.7 Hz, 4H), 1.53-1.43 (m, 6H); LC-MS (ESI, m/z) = 451.1 (M+H ⁺ ).

3. #14-6, #14-7, #14-8 synthesis scheme

#14-6: (E)-N-ethyl-1-(3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)phenyl)methanimine oxide

Compound 3 (100 mg, 0.32 mmol) obtained from Preparation Example 2 was placed in a reaction flask and dissolved in 3 mL of anhydrous EtOH. N-methylhydroxylamine·HCl (79 mg, 0.95 mmol) and potassium acetate (93 mg, 0.95 mmol) were added and stirred at room temperature for 24 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 76 mg of the title compound #14-6. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.35 (s, 1H), 8.12 (s, 1H), 7.86 (d, 2H, J = 8.59 Hz), 7.74 (d, 2H, J = 8.59 Hz) , 7.70 (m, 1H), 7.56 (m, 1H), 7.44 (t, 1H, J = 7.63 Hz), 7.32 (m, 1H), 3.98 (q, 2H, J=9.00 Hz), 1.56 (t, 3H, J=9.00 Hz); LC-MS (ESI, m/z) = 360.2 (M+H ⁺ ).

#14-7: (E)-N-isopropyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide

Compound 3 (100 mg, 0.32 mmol) obtained from Preparation Example 2 was placed in a reaction flask and dissolved in 3 mL of anhydrous EtOH. N-isopropylhydroxylamine·HCl (105 mg, 0.95 mmol) and potassium acetate (93 mg, 0.95 mmol) were added and stirred at room temperature for 24 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 60 mg of the title compound #14-7. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.32 (s, 1H), 8.12 (s, 1H), 7.86 (d, 2H, J = 8.50 Hz), 7.71 (d, 2H, J = 8.50 Hz) , 7.65 (m, 1H), 7.56 (m, 1H), 7.41 (t, 1H, J = 7.61 Hz), 7.29 (m, 1H), 4.21 (m, 1H), 1.51 (s, 3H), 1.49 ( s, 3H); LC-MS (ESI, m/z) = 374.0 (M+H ⁺ ).

#14-8: (E)-N-tert-butyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide

Compound 3 (100 mg, 0.32 mmol) obtained from Preparation Example 2 was placed in a reaction flask and dissolved in 3 mL of anhydrous EtOH. N-tert-butylhydroxylamine·HCl (118 mg, 0.95 mmol) and potassium acetate (93 mg, 0.95 mmol) were added and stirred at room temperature for 24 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 71 mg of the title compound #14-8. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.38 (s, 1H), 8.21 (s, 1H), 7.84 (d, 2H, J = 8.12 Hz), 7.72 (d, 2H, J = 8.12 Hz) , 7.59 (m, 1H), 7.53 (m, 1H), 7.40 (t, 1H, J = 7.61 Hz), 7.27 (m, 1H), 1.60 (s, 9H); LC-MS (ESI, m/z) = 388.1 (M+H ⁺ ).

4. #14-9, #14-10 synthesis scheme

#14-9: (E)-3-(1-(4-( trifluoromethyl )phenyl)-1H- pyrazol -4-yl)benzaldehyde O-ethyl oxime

Compound 3 (100 mg, 0.32 mmol) obtained from Preparation Example 2 was placed in a reaction flask and dissolved in 5 mL of anhydrous DCM. After adding ethoxyamine·HCl (46 mg, 0.47 mmol) and pyridine (56 μL, 0.70 mmol), it was stirred at room temperature for 5 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 78 mg of the title compound #14-9. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.46 (s, 1H), 8.32 (s, 1H), 8.08 (s, 1H), 7.80 (d, 2H, J = 8.45 Hz), 7.71 (d, 2H, J = 8.45 Hz), 7.64 (m, 1H), 7.53 (m, 1H), 7.41 (t, 1H, J = 7.61 Hz), 7.32 (m, 1H), 3.57 (q, 2H, J=9.82 Hz), 1.51 (t, 3H, J=9.82 Hz); LC-MS (ESI, m/z) = 360.1 (M+H ⁺ ).

#14-10: (E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-( tert -butyl) oxime

Compound 3 (100 mg, 0.32 mmol) obtained from Preparation Example 2 was placed in a reaction flask and dissolved in 5 mL of anhydrous DCM. tert-butoxyamine·HCl (59 mg, 0.47 mmol) and pyridine (56 μL, 0.70 mmol) were added and stirred at room temperature for 5 hours. The reaction product was confirmed by TLC, then diluted with EtOAc and washed with purified water. The separated organic layer was dehydrated with anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and separated by column chromatography to obtain 85 mg of the title compound #14-10. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.52 (s, 1H), 8.38 (s, 1H), 8.12 (s, 1H), 7.83 (d, 2H, J = 8.42 Hz), 7.74 (d, 2H, J = 8.42 Hz), 7.63 (m, 1H), 7.50 (m, 1H), 7.47 (t, 1H, J = 7.59 Hz), 7.32 (m, 1H), 1.25 (s, 9H); LC-MS (ESI, m/z) = 388.0 (M+H ⁺ ).

< Example 2> Breast cancer cell lines and their culture methods

Human-derived tumor cell lines and breast cancer cell lines MCF7, T47D, BT549, MDA-MB231, and MDA-MB453 contain 10% fetal bovine serum (FBS; HyClone, South Logan, UT) and 1% penicillin/streptomycin. Cultured in DMEM medium (Corning, NY, USA) at 37°C and 5% CO ₂ conditions.

Thereafter, the cell lines and compounds prepared through the above method were used in the examples below.

< Example 3> KRCT Confirmation of cancer cell-specific killing effect (anti-cancer effect) of -1

One. KRCT Confirmation of cancer cell killing effect (anti-cancer effect) through cell proliferation and survival analysis of -1

To confirm the inhibition of cancer cell proliferation by KRCT-1, WST-8 analysis (Cyto XTM cell viability assay kit, LPS solution, Daejeon, Korea) was performed according to the manufacturer's protocol. Colon cancer (DLD1, HCT-116), lung cancer (H1299, A549), brain tumor (U373, LN18), breast cancer (T47D, BT549, MDA-MB 453, MCF7 and MDA MB-231), stomach cancer (SNU668), blood cancer (MV4-11) cells were seeded in a 96-well plate, cultured for 24 hours, and then treated with the above compounds. Afterwards, the absorbance was measured at 450 nm using a Versamax microplate reader (Molecular Devices, CA, USA), and the results of the experiment are shown in Figures 2 and 3B.

2. Confirmation of cancer cell killing effect (anti-cancer effect) through clonogenic assay of KRCT-1

Colony formation analysis was performed to determine whether the selected KRCT-1 could efficiently kill cancer cells. First, cells of various breast cancer cell lines BT549, MCF7, T47D and MDA MB231 (500-1000 cells) were cultured in a 60 mm culture plate, and after 24 hours, they were treated with 10 μM each of the above compounds and cultured for 14 days. It was fixed and stained with tryptophan blue supplemented with methanol, and quantified using Image J software, which is shown in Figure 3C.

As confirmed in Figure 3C, it was confirmed that the number and size of haemocyte colonies were significantly reduced upon treatment with KRCT-1 in various breast cancer cell lines. As a result, KRCT-1 effectively kills cancer cell colonies.

3. Confirmation of cancer cell killing effect (anti-cancer effect) through tumor sphere forming assay of KRCT-1

A tumor sphere formation assay was performed to determine whether the selected KRCT-1 could effectively reduce the formation of tumorspheres of cancer cells. Breast cancer cells MCF7 (2500-10000 cells) were grown on low-adhesion plates with 1% penicillin, B-27 (1:50; invitrogen), 20 ng/mL epidermal growth factor (R&D Systems), and 20 ng/mL fibroblasts. The cells were cultured by suspending them in serum-free DMEM/F12 (welGENE) containing cell growth factors (R&D Systems). After 24 hours, the cells were treated with the compound and cultured for 14 days. The diameter of the tumor spheres was measured and quantified, and this is shown in Figure 3D.

As confirmed in Figure 3D, it was confirmed that the diameter of cancer cell tumor spheres was significantly reduced upon treatment with KRCT-1 in various breast cancer cell lines. As a result, KRCT-1 effectively reduces the formation of tumor spheres in cancer cells.

< Example 4> Immunodeficiency mouse tumor model ( Xenograft mouse model) KRCT Confirmed anti-cancer effect of -1

We attempted to verify whether the anticancer effect of the compound actually occurs at the biological level by subcutaneously transplanting cancer cell lines into an immunodeficient mouse tumor model. Tumors were formed by transplanting the BT549 cell line subcutaneously into the thighs of immunodeficient mice, and when the tumor size reached 50-100 mm ³ , the compound was intraperitoneally injected at 30 mg/kg twice a week. Tumor volume and mouse weight were measured three times a week for the control group and the KRCT-1 treatment group, respectively. In Figure 4A and Figure 4B, it was observed that at 4 weeks after transplantation of breast cancer cell lines, the tumor volume and size of the KRCT-1 treated group was smaller than that of the control group. Western blotting was performed on the tumor growth antigen Ki-67 on the isolated mouse tumor, and is shown in B of Figure 4b. Figure 4A (B) shows the results of measuring the weight of the mouse three times a week while administering the compound.

As shown in Figure 4, the group treated with KRCT-1 had smaller tumor volume and size, and Ki-67 expression was clearly reduced, indicating that the KRCT-1 compound inhibited tumor growth and had an anticancer effect. It was confirmed that it exists. Additionally, there was no significant change in the body weight of immunodeficient mice upon treatment with KRCT-1, confirming that KRCT-1 does not exhibit toxicity and can inhibit tumor growth.

< Example 5> KRCT Confirmation of cancer cell killing effect of -1

One. FACS Confirmation of cell death effect through

After Annexin V/PI staining, FACS analysis was performed to confirm the cancer cell killing effect. Cancer cell lines MCF7 and BT549 were treated in the presence or absence of KRCT-1 20 μM for 48 h, and Annexin V binding was analyzed using the Annexin V-FITC apoptosis Detection Kit (BD Biosciences, San Jose, USA) according to the manufacturer's recommended protocol. CA, USA) was used. Briefly, all cells ^, including the cell medium, were collected, centrifuged at 1500 rpm for 5 minutes, washed with 1 Added to the tube, 5 μl of Annexin V-FITC and 5 μl of PI were added and incubated at RT for 15 minutes in the dark. 400 μl of 1× binding buffer was added to each tube, and samples were analyzed by flow cytometry. The results of the FACS assay are shown in Figure 5A. As a result, it was confirmed that MCF7 induced about 40% cell death and BT549 induced about 23% cell death.

2. Confirmation of apoptosis effect through confirmation of molecular expression

Cell death caused by KRCT-1 was confirmed through Western blotting experiments. 20 μg of protein was separated through SDS-PAGE electrophoresis and transferred to a PVDF membrane, and the following specific antibodies were used: rabbit polyclonal antibody cleaved PARP (Asp214) (#9541) and phospho-Chk2 (Thr68) ( #2661; Cell Signaling Technology, MA); Rabbit monoclonal antibodies caspase3 (#9664), chk2 (#6334S), survivin (#2808S); Mouse monoclonal antibody caspase 9 (551246; BD Pharmingen, CA, USA). In addition, the mouse monoclonal antibody β-actin (C4; Santa Cruz Biotechnology, CA, USA) was used as a loading control, washed with 1 The binding protein was detected using and an image of the band was obtained using an Amersham Imager 600 system (GE Healthcare Life Science), which is shown in Figure 5b.

Breast cancer cell lines BT549, T47D, and MCF7 were treated with KRCT-1 at doses of 0, 2.5, 5, 10, and 20 μM each for 24 h in a dose-dependent manner, and the expression level of survivin, which is essential for cancer cell survival, and clv-PARP, an apoptosis marker, were measured. , the expression levels of caspase 9 and caspase 3 were confirmed. Depending on the concentration of treated KRCT-1, the expression levels of survivin and pro-caspase 9 present in cancer cells were reduced, and cleaved PARP and caspase 3 were confirmed to increase during apoptosis. Through the above results, it was confirmed that cell death was activated due to KRCT-1.

< Example 6> KRCT Through processing of -1 cancer cell line Confirmation of increased radiation sensitivity effect

One. fungus Confirmation of radiation combination effect through formation analysis method

A colony formation assay was performed to confirm the effect of combined use of KRCT-1 and radiation. To perform the colony formation assay, breast cancer cell lines BT549, T47D and MCF7 were treated with 2.5 μM of KRCT-1 compound and treated with 2.5 Gy radiation using Biobeam GM8000 (Gamma-Service Medical GmbH, Leipzig, Germany). The remaining experimental technique for colony formation was performed in the same manner as in the above example, and is shown in Figure 6a. In the group treated with KRCT-1, there was a significant radiation treatment effect on both cancer cell lines compared to the group treated with DMSO, and the number of cancer cell colonies was confirmed to be reduced.

2. western Blotting Confirmation of radiation combined effect through experiment

In order to confirm the effect of increasing the radiation sensitivity of cancer cell lines through the treatment of KRCT-1 through Western blotting experiments, breast cancer cell lines BT549, T47D and MCF7 were treated with 10 μM of the KRCT-1 compound and incubated with Biobeam GM8000 (Gamma - Gamma-Service Medical GmbH, Leipzig, Germany) and cultured for 30 h after treatment with 3 Gy radiation. To perform Western blotting, 20 μg of protein was separated through SDS-PAGE electrophoresis, transferred to PVDF membrane, and the following specific antibodies were used: rabbit polyclonal antibody phospho-Chk2 (Thr68) (#2661; Cell Signaling Technology, MA); Rabbit monoclonal antibody chk2 (#6334S) was used. The remaining conditions for Western blotting were performed the same as in the above example. As a result, as shown in Figure 6A, the expression level of survivin decreased and the expression level of phosphorylated Chk2 increased in the group treated with KRCT-1 after radiation treatment. Through the above results, it was confirmed that cancer cell death was effective.

3. KRCT Check the effect of -1 on DNA damage

To determine how KRCT-1 affects DNA damage induced by radiation, a γ-H2AX foci experiment was performed. When single or double strands of DNA are broken by various DNA damaging agents, including radiation, γ-H2AX phosphorylation occurs and nuclear foci are created in the cell nucleus. Using this as an indicator, the effect of combination according to DNA damage was evaluated. 2.5 × 10 ⁴ MCF7 cells were cultured in a 24-well plate for 48 h, treated with KRCT-1 20 μM for 24 h, and 1 h after irradiation with 2 Gy of radiation, the cells were fixed. After washing with phosphate-buffered saline for permeabilization, 4% paraformaldehyde and 0.2% Triton-X were added and reacted for 10 min each. Pretreatment with 5% fetal calf serum for 1 h, followed by treatment with 1:100 diluted γ-H2AX (05-656; EMD Millipore Corp., MA, USA) antibody against phosphorylated H2AX for 16 h, followed by 0.1% Triton Washed three times with 100/phosphate buffered saline solution (Tritonx-100/PBS). They were labeled with Alexa488-conjugated secondary antibody (AP132JA4, Invitrogen) at room temperature for 2 h, and phosphorylated γ-H2AX foci were imaged and analyzed with IN Cell Analyzer 6000 (GE Healthcare Life Science). As a result, as shown in Figure 6B, it was confirmed that DNA damage occurred in a relatively large number of cancer cells when treated with KRCT-1 and radiation compared to each control group, and that sensitivity to radiation was improved. Taken together, it was confirmed that KRCT-1 is a substance that can be effectively used as a radiosensitizer in cancer cells.

< Example 7> Confirmation of Trk-A selective inhibition activity of KRCT-1: In vitro Efficacy and specificity testing through kinase assay

To measure the selective reactivity of KRCT-1 to a specific kinase, in vitro kinase assay profiling was requested from Goma Biotech (Seoul, Korea), and enzyme reaction experiments were performed at Eurofins Pharma Discovery Services (Scotland, UK). . For kinase profiling, a broad oncology panel of 104 kinase series and 10 μM KRCT-1 were used in the reaction. To obtain the IC ₅₀ kinase assay of Trk-A, KRCT-1 serially diluted 3 times from 100 μM was used.

< Example 8> In cancer cell lines KRCT by -1 Trk -A confirmation of inhibitory activity

It was confirmed whether KRCT-1 inhibits downstream signal transduction by inhibiting the activity of Trk-A kinase in cancer cell lines. When Trk-A is activated by its ligand, the MEK/ERK and STAT3 signaling pathways are activated within the cell. Using this as an indicator, changes in phosphorylated ERK and phosphorylated STAT3 were confirmed when cancer cell lines were treated with KRCT-1 concentrations (2.5, 5, 10, and 20 μM). To perform Western blotting, 20 μg of protein was separated by SDS-PAGE electrophoresis, transferred to PVDF membrane, and the following specific antibodies were used: rabbit polyclonal antibody phospho-STAT3 (Tyr705) (ab76315; Abcam, UK), mouse monoclonal antibodies phospho-ERK (Thr202/Tyr204) (sc-514302; Santa Cruz Biotechnology), ERK (sc-7383; Santa Cruz Biotechnology), and STAT3 (sc8019; Santa Cruz Biotechnology) were used. The remaining conditions for Western blotting were performed the same as in the above example.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (12)

A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Formula 1]

In Formula 1,
R ₁ is selected from hydrogen, (C1~C6)alkyl, (C2~C6)alkene, (C2~C6)alkyne, substituted or unsubstituted aryl(C1~C6)alkyl, or substituted or unsubstituted aryl, wherein The aryl may be substituted with 1 to 3 substituents and is independently selected from the group consisting of halo, nitro, cyano, hydroxy and oxo, selected from the group consisting of fluoro, chloro and bromo,
R ₂ is selected from hydrogen, (C1~C6)alkylamino or di(C1~C6)alkylamino,
A is any of aryl, pyridine, thiophene, or biphenyl,
_{and ​ ​} A compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof.
[Formula 2]

In Formula 2,
Y is hydrogen, CF ₃ , CHF ₂ , CH ₂ F, (C1~C6)alkyl, (C1~C6)alkoxy, or halo selected from the group consisting of fluoro, chloro, and bromo,
n is an integer from 0 to 5,
R is or and R ₁ and R ₂ are each independently selected from hydrogen or substituted or unsubstituted (C1~C6)alkyl, where the alkyl may be substituted with 1 to 3 substituents, and are independently selected from fluoro, chloro and broro. selected from halo or hydroxy selected from the group consisting of The method of claim 1, wherein the compound represented by Formula 1 is N-methyl-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ethane Sulfonamide [N-methyl-N-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol -4-yl)phenyl)ethanesulfonamide], N-(4-chlorobenzyl)-N-(3 -(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide[N-(4-chlorobenzyl)-N-(3-(1-(4- (trifluoromethyl) phenyl)-1H-pyrazol-4-yl)phenyl)ethanesulfonamide], N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazole -4-yl)phenyl)azanesulfonamide [N,N-dimethyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N ,N-dimethyl-N'-methyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N -dimethyl-N'-methyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(4 -Chlorobenzyl)-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N-dimethyl-N' -(4-chlorobenzyl)-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3- (1-(4-phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N-dimethyl-N'-(3-(1-(4-phenyl)-1H-pyrazol- 4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3-(1-(4-(methyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide [N ,N-dimethyl-N'-(3-(1-(4-(methyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3-(1 -(4-(methoxy)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-(3-(1-(4-(methoxy)phenyl) -1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3-(1-(pyridin-4-yl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide Fonamide [N,N-dimethyl-N'-(3-(1-(pyridin-4-yl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3 -(1-(thiophen-4-yl)-1H-pyrazol-4-yl)phenyl)azansulfonamide[N,N-dimethyl-N'-(3-(1-(thiophen-4-yl) )-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)phenyl)ah Zansulfonamide [N,N-dimethyl-N'-(3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-(3- (1-(4-([1,1'-biphenyl]-1H-pyrazol-4-yl)phenyl)azanesulfonamide[N,N-dimethyl-N'-(3-(1-(4 -([1,1'-biphenyl]-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-isobutyl-N'-(3-(1-(4-(tri Fluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azansulfonamide [N,N-dimethyl-N'-isobutyl-N'-(3-(1-(4-(trifluoromethyl)phenyl )-1H-pyrazol-4-yl)phenyl)azanesulfonamide], N,N-dimethyl-N'-allyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol Zol-4-yl) phenyl) azansulfonamide [N, N-dimethyl-N'-allyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl) phenyl)azanesulfonamide], N,N-dimethyl-N'-phenyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide Fonamide [N,N-dimethyl-N'-phenyl-N'-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)azanesulfonamide] and N-(3- (1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)piperidine-1-sulfonamide [N-(3-(1-(4-(trifluoromethyl)phenyl) )-1H-pyrazol-4-yl)phenyl)piperidine-1-sulfonamide], or a pharmaceutically acceptable salt thereof. The method of claim 2, wherein the compound represented by Formula 2 is (E)-N-ethyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl ) Phenyl) methanimine oxide [(E)-N-ethyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide], (E)- N-isopropyl-1-(3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide[(E)-N-isopropyl-1- (3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide], (E)-N-tert-butyl-1-(3-(1-(4- (trifluoromethyl)phenyl)-1H-pyrazol-4-yl)phenyl)methanimine oxide [(E)-N-tert-butyl-1-(3-(1-(4-(trifluoromethyl)phenyl) -1H-pyrazol-4-yl)phenyl)methanimine oxide], (E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-ethyl oxime [(E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-ethyl oxime] and (E)-3-(1-(4-(trifluoromethyl) methyl)phenyl)-1H-pyrazol-4-yl)benzaldehyde O-(tert-butyl)oxime [(E)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl) benzaldehyde O-(tert-butyl) oxime], or a pharmaceutically acceptable salt thereof. A pharmaceutical composition for preventing or treating cancer containing the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient. The method of claim 5, wherein the cancer is brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, nasal cavity/sinus. Cancer, nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, stomach cancer, liver cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer. , small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, male genital tract tumor, penile cancer, urethral cancer, prostate cancer, female genital tract tumor, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female external reproductive organs. A pharmaceutical composition for preventing or treating cancer, characterized in that the cancer is selected from the group consisting of primary cancer, female urethral cancer, skin cancer, myeloma, leukemia, and malignant lymphoma. The pharmaceutical composition for preventing or treating cancer according to claim 5, wherein the pharmaceutical composition is administered in combination with radiation during cancer treatment. The pharmaceutical composition for preventing or treating cancer according to claim 5, wherein the pharmaceutical composition increases the sensitivity to radiation of cancer cells or cancer cells with radioresistance. A health functional food composition for preventing or improving cancer containing the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for radiotherapy sensitization for cancer containing the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient. The method of claim 10, wherein the cancer is brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, nasal cavity/sinus. Cancer, nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, stomach cancer, liver cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer. , small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, male genital tumor, penile cancer, urethral cancer, prostate cancer, female genital cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female external reproductive organs. A pharmaceutical composition for radiotherapy sensitization for cancer, characterized in that the cancer is selected from the group consisting of primary cancer, female urethral cancer, skin cancer, myeloma, leukemia, and malignant lymphoma. The pharmaceutical composition for radiation therapy sensitization for cancer according to claim 10, wherein the pharmaceutical composition is administered in combination with radiation during cancer treatment.