Composition for preventing, improving, or treating cancer

Abstract

The composition of the present invention can not only very effectively inhibit the growth of cancer cells, but also very effectively inhibit the metastasis of cancer cells to other tissues, and therefore can be very effectively used to prevent, improve, or treat cancer, as well as inhibit cancer metastasis.

Description

Technical Field

[0001] The present invention relates to a composition for preventing, improving or treating cancer. Background Art

[0002] Cancer is a fatal disease that causes tissue cells to proliferate abnormally and indefinitely, forming tumors that prevent organs from performing their normal functions, thus threatening the patient's life. In 2017, the leading cause of death in South Korea was malignant tumors (cancer), accounting for 27.6% of the total number of deaths. Although the location of cancer varies by race, gender, and region, in most countries, prostate cancer is the most common among men, while breast cancer is the most common among women. In particular, breast cancer is the highest cause of death among female cancer patients worldwide. This is because breast cancer can be caused by a variety of factors such as environmental and genetic factors, and its incidence is increasing year by year.

[0003] Paclitaxel (Taxol) is an existing FDA-approved anticancer drug that is used to treat ovarian cancer, breast cancer, lung cancer, etc. Its mechanism of action is to inhibit cell division by binding to cell spindles, thereby inhibiting the growth of disorderly proliferating tumor cells. Therefore, the use of this anticancer drug can cause various adverse reactions, such as hair loss, muscle pain and diarrhea. However, even with these adverse reactions, this anticancer preparation has been continuously used in clinical practice because it can effectively inhibit tumor cell division.

[0004] At the same time, in malignant tumors where general anticancer drugs do not show therapeutic effects, the following phenomenon is often observed: tumor cells developed in a specific organ (primary organ) of the patient will acquire new genetic characteristics necessary for metastasis as the cancer progresses, and then invade blood vessels and lymph nodes, spread from the primary organ along with the lymph, deposit in the tissues of other organs, and then begin to proliferate. Such malignant tumors can be treated with local therapy and systemic therapy. Local treatment methods such as therapeutic surgery or radiotherapy can be applied to malignant tumors known to have severe local symptoms caused by tumor metastasis or malignant tumors known to be able to improve their clinical course by surgical treatment. However, except for a very small number of tumors such as lymphoma, the cure rate of other malignant tumors that have metastasized is very low.

[0005] Therefore, there is an urgent need to develop an anticancer drug that can prevent tumor cells from metastasizing to organs such as the lungs, while overcoming the shortcomings of anticancer drugs developed for cancer treatment so far, such as low specificity for cancer cells and many adverse reactions. Summary of the invention

[0006] Technical issues

[0007] An object of the present invention is to provide a pharmaceutical composition, a food composition and a cosmetic composition for preventing, improving or treating cancer.

[0008] Another object of the present invention is to provide a pharmaceutical composition, a food composition and a cosmetic composition for inhibiting cancer metastasis.

[0009] Another object of the present invention is to provide a method for preventing or treating cancer.

[0010] Another object of the present invention is to provide a method for inhibiting cancer metastasis.

[0011] However, the objectives to be achieved by the present invention are not limited to the above objectives, and a person skilled in the art can clearly understand other objectives not mentioned herein through the following description.

[0012] Technical Solutions

[0013] One embodiment of the present invention provides a composition for preventing, improving, or treating cancer, wherein the composition contains a compound represented by the following formula 1 as an active ingredient:

[0014] [Formula 1]

[0015]

[0016] in:

[0017] L1 and L2 are each independently selected from the following group: C3-C 40 Cycloalkylene, C6-C 60 Arylene, and heteroarylene having 5 to 60 nuclear atoms;

[0018] X and Y are each independently selected from the group consisting of deuterium, halogen, cyano, nitro, sulfonyl, C1-C 10 Alkylsulfonyl, azide group, hydroxyl, C1-C 40 Alkyl, C2-C 40 Alkenyl, C1-C 40 Alkoxy, unsubstituted or substituted C6-C 60 Aryloxy, unsubstituted or substituted C3-C 40 Cycloalkyl, unsubstituted or substituted heterocycloalkyl having 3 to 20 nuclear atoms, unsubstituted or substituted C6-C 60 Aryl, unsubstituted or substituted heteroaryl having 5 to 60 nuclear atoms, and -NR'R";

[0019] R' and R" are each independently selected from the group consisting of hydrogen, C1-C 10 Alkyl, C6-C 60 Aryl, C3-C 40 Cycloalkyl, C6-C60 Arylsulfonyl, and heteroaryl having 5 to 60 nuclear atoms;

[0020] n and m are each independently an integer from 0 to 5, provided that n and m cannot be 0 at the same time;

[0021] When there are multiple X or Y, the multiple X or Y may be the same as or different from each other; and

[0022] The arylsulfonyl group in each of R' and R" may be unsubstituted or substituted with at least one substituent selected from the group consisting of deuterium, halogen, and nitro.

[0023] The compounds of the present invention shown in Formula 1 can very effectively inhibit uncontrolled cancer cell proliferation (also known as neoplasia). In view of the fact that the composition of the present invention can cause the death (apoptosis) of uncontrolled cells and may inhibit tumor growth by inhibiting cell growth, it can be very effectively used to prevent, improve, or treat tumors.

[0024] The term "aryl" as used herein refers to a C6-C8 group derived from a single ring or two or more rings bonded to each other. 40 A monovalent substituent of an aromatic hydrocarbon. In addition, an aryl group may also include two or more rings that are pendant (e.g., simply connected) or fused to each other. Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, phenanthrenyl, and anthracenyl.

[0025] As used herein, the term "arylene" refers to a group of atoms obtained by removing a hydrogen atom from aromatic hydrocarbons, and also includes those with fused rings or those in which two or more independent benzene rings or fused rings are directly or bonded by groups such as vinylene. The arylene can have any substituent described herein, for example, alkoxy, and the carbon number of this part except the substituent is generally 6-60. In addition, the total carbon atom of the arylene including the substituent is generally 6-100. The example of such arylene includes, but is not limited to, phenylene, naphthalene diyl, dimethoxybenzyl, anthracene diyl, biphenyl diyl, terphenyl diyl, fused compound group, fluorene diyl, stilbene diyl, distyryl diyl, benzofluorene diyl, dibenzofluorene diyl, and the like.

[0026] As used herein, the term "heteroarylene" refers to a divalent monocyclic aromatic group or a divalent polycyclic aromatic group, wherein at least one aromatic ring contains one or more heteroatoms independently selected from O, S or N. Each ring of the heteroarylene group may contain one or two O atoms, one or two S atoms, and / or 1-4 N atoms, provided that the total number of heteroatoms in each ring is 4 or less and each ring contains at least one carbon atom. Examples of heteroarylene groups include, but are not limited to, benzofuranylene, benzimidazolylene, benzisoxazolylene, benzopyranylene, benzothiadiazolylene, benzothiazolylene, benzothiophenylene, benzotriazolylene, benzoxazolylene, furopyridylene, imidazopyridylene, imidazothiazolylene, indolizinylene, indolylene, indazolylene, isobenzofuranylene, isobenzothiophenylene, isoindolylene, isoquinolylene, isothiazolylene, naphthyridinylene, oxazolopyridylene, phthalazinylene, pteridinylene, furinylene, pyridopyridineene, pyrrolopyridylene, quinolylene, quinoxalylene, quinazolinylene, thiadiazolopyrimidylene, and thienopyridylene. Examples of tricyclic heteroarylene groups include, but are not limited to, acridinylene, benzindolylene, carbazolylene, dibenzofuranylene, perimidinylene, phenanthrolinylene, phenanthridinylene, phenarsazinylene, phenazinylene, phenothiazinylene, phenoxazinylene, and the like.

[0027] As used herein, the term "alkyl" refers to a linear or branched saturated monovalent hydrocarbon group, wherein the alkyl group may be optionally substituted with one or more substituents herein. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (including all isomeric forms thereof), n-propyl, isopropyl, butyl (including all isomeric forms thereof), n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl (including all isomeric forms thereof), and hexyl (including all isomeric forms thereof).

[0028] As used herein, the term "alkylsulfonyl" includes methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, tert-butylsulfonyl, and the like. The number of carbon atoms constituting the alkylsulfonyl group is preferably 1-10, but is not limited thereto.

[0029] As used herein, the term "alkenyl" refers to a straight or branched monovalent hydrocarbon group containing one or more carbon-carbon double bonds, wherein the number of carbon-carbon double bonds is 1-5 in one embodiment and 1 in another embodiment. The alkyl group may be optionally substituted with one or more substituents herein. As known to those skilled in the art, the term "alkenyl" includes groups having a "cis" or "trans" structure or a combination thereof, or having a "Z" or "E" structure or a combination thereof. Examples of alkenyl groups include, but are not limited to, vinyl, propene-1-yl, propene-2-yl, allyl, butenyl, and 4-methylbutenyl.

[0030] As used herein, the term "aryloxy" is a monovalent substituent as shown in RO-, where R represents an aryl group having 5 to 40 carbon atoms. Examples of aryloxy include, but are not limited to, phenoxy, naphthoxy, diphenoxy, and the like.

[0031] As used herein, the term "heterocycloalkyl" refers to a monovalent monocyclic ring system having 3-20 ring atoms, wherein 1-3 heteroatoms are selected from N, O, P or S, and the remaining ring atoms are C. One or more hydrogen atoms in the heterocycloalkyl group may be optionally substituted. Examples of the heterocycloalkyl group include, but are not limited to, pyrrolidinyl, pyrazolyl, imidazolidinyl, piperidinyl and piperazine.

[0032] As used herein, the term "alkoxy" is a monovalent substituent as shown by R'O-, wherein R' represents an alkyl group having 1-40 carbon atoms, which may include a straight chain, a straight chain or a ring structure. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, 1-propoxy, tert-butoxy, n-butoxy, pentoxy, and the like.

[0033] As used herein, the term "arylamine" refers to an amine substituted with an aryl group having 6 to 40 carbon atoms.

[0034] As used herein, the term "cycloalkyl" is a monovalent substituent derived from a monocyclic or polycyclic non-aromatic hydrocarbon having 3 to 40 carbon atoms. Examples of such cycloalkyls include, but are not limited to, cyclopropyl, cyclopentyl, cyclohexyl, norbornyl, adamantane, and the like.

[0035] As used herein, the term "halogen" refers to fluorine, chlorine, bromine and / or iodine.

[0036] As used herein, the terms "substituted alkyl", "substituted alkylene", "substituted heteroalkylene", "substituted alkenyl", "substituted alkenylene", "substituted heteroalkenylene", "substituted alkynyl", "substituted alkynylene", "substituted cycloalkyl", "substituted heterocycloalkyl", "substituted cycloalkylene", "substituted aryl", "substituted aryloxy", "substituted arylene", "substituted aralkyl", "substituted heteroaryl", "substituted heteroarylene", "substituted heterocycle", or "substituted heterocyclylene" mean that the substituted alkyl, substituted alkynyl, substituted alkynylene, substituted cycloalkyl, substituted heterocycloalkyl, substituted cycloalkylene, substituted aryl, substituted aryloxy, substituted arylene, substituted aralkyl, substituted heteroaryl, substituted heteroarylene, substituted heterocycle, or substituted heterocyclylene can be further substituted by one or more substituents, for example, substituents independently selected from the following group: C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Cycloalkyl, C 6-14 Aryl, C 7-15 Aralkyl, heteroaryl, heterocyclic, hydroxy, oxo (=O), halogen, cyano (-CN), nitro (-NO2), -C(O)R a 、-C(O)OR a 、-C(O)NR b R c 、-C(NR a )NR b R c 、-OR a 、-OC(O)R a 、-OC(O)OR a 、-OC(O)NR b R c 、-OC(=NR a )NR b R c 、-OS(O)R a 、-OS(O)2R a 、-OS(O)NR b R c 、-OS(O)2NR b R c 、-NR b R c 、-NR a C(O)R d 、-NR a C(O)OR d 、-NRa C(O)NR b R c 、-NR a C(=NR d )NR b R c 、-NR a S(O)R d 、-NR a S(O)2R d 、-NR a S(O)NR b R c 、-NR a S(O)2NR b R c 、-SR a 、-S(O)R a 、-S(O)2R a 、-S(O)NR b R c , and -S(O)2NR b R c , where R a , R b , R c , and R d may each independently be (a) hydrogen; or (b) C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Cycloalkyl, C 6-14 Aryl, C 7-15 aralkyl, heteroaryl, or heterocyclic; or (c) R b and Rc may form a substituted heterocyclic group together with the nitrogen atom to which they are attached.

[0037] In the present invention, the compound represented by Formula 1 may be, but is not limited to, the compound represented by the following Formula 2:

[0038] [Formula 2]

[0039]

[0040] in

[0041] n, m, X and Y are each as defined in Formula 1 above.

[0042] In a preferred embodiment of the present invention, n and m in Formula 2 may be an integer of 1 or 2, respectively, independently.

[0043] X and Y can be independently selected from the following groups: sulfonyl, C1-C 10 Alkylsulfonyl, C1-C 40Alkoxy, -NR'R", hydroxyl, C6-C 60 Aryloxy, and unsubstituted or substituted heterocycloalkyl having 3 to 20 nuclear atoms;

[0044] R' and R" can each independently be hydrogen or C6-C 60 an arylsulfonyl group, and

[0045] The arylsulfonyl group in each of R' and R" may be unsubstituted or substituted with at least one halogen.

[0046] In a preferred embodiment of the present invention, in Formula 2,

[0047] n can be 1;

[0048] X may be -NR'R" or a substituent shown in Formula 3 below; and

[0049] R' and R" may each independently be hydrogen or a substituent represented by the following formula 4:

[0050] [Formula 3]

[0051]

[0052] [Formula 4]

[0053]

[0054] in

[0055] R1 can be selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, C1-C 40 Alkyl, and C2-C 40 alkenyl, and preferably, R1 may be hydrogen or hydroxyl; and

[0056] R2 may be selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, and nitro, and is preferably halogen, more preferably chlorine, but is not limited thereto.

[0057] In a preferred embodiment of the present invention, in the above formula 2,

[0058] Y may be, but is not limited to, a hydroxyl group, a C1-C6 alkoxy group, or a substituent shown in the following formula 5:

[0059] [Formula 5]

[0060]

[0061] In a preferred embodiment of the present invention, in Formula 2,

[0062] n can be 1;

[0063] X can be -NR'R";

[0064] R' and R" may each independently be hydrogen or a substituent as shown in Formula 5 above:

[0065] m can be 2;

[0066] Y can be, but is not limited to, a substituent of a C1 alkoxy group:

[0067] In the present invention, the compound represented by Formula 1 may be, but is not limited to, any compound selected from the following group:

[0068]

[0069]

[0070] In another embodiment of the present invention, the compound may be, but is not limited to, the following compounds:

[0071]

[0072] In another embodiment of the present invention, the compound may be, but is not limited to, the following compounds:

[0073]

[0074] The cancer described herein can be at least one selected from the group consisting of breast cancer, colorectal cancer, lung cancer, liver cancer, gastric cancer, esophageal cancer, pancreatic cancer, gallbladder cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, colon cancer, cervical cancer, endometrial cancer, choriocarcinoma, skin cancer, ovarian cancer, thyroid cancer, brain cancer, blood cancer, head and neck cancer, malignant melanoma, and lymphoma, and preferably can be breast cancer, but is not limited thereto.

[0075] The breast cancer described herein may be, but is not limited to, triple-negative breast cancer.

[0076] The term "triple-negative breast cancer" as used herein refers to breast cancer that is negative for estrogen receptor, progesterone receptor, and Her2 receptor, is prone to recurrence and metastasis, and cannot be cured by conventional targeted therapeutic agents for breast cancer.

[0077] As used herein, the term "prevention" may include, but is not limited to, any effect of blocking, inhibiting or delaying symptoms caused by the uncontrolled growth of cancer cells by using the composition of the present invention.

[0078] As used herein, the term "improvement" may include, but is not limited to, any effect of alleviating or beneficially changing symptoms caused by uncontrolled growth of cancer cells, etc., by using the composition of the present invention.

[0079] As used herein, the term "treatment" may include, but is not limited to, any effect of alleviating or beneficially changing the symptoms caused by the uncontrolled growth of cancer cells, etc., by using the composition of the present invention.

[0080] Another embodiment of the present invention provides a composition for inhibiting cancer metastasis, wherein the composition contains a compound represented by the following formula 1 as an active ingredient:

[0081] [Formula 1]

[0082]

[0083] wherein n, m, X and Y are respectively as defined in Formula 1 of the composition for preventing, ameliorating or treating cancer.

[0084] In the present invention, "cancer metastasis" refers to a phenomenon in which tumor cells developed in the primary organ acquire new genetic characteristics necessary for metastasis as the cancer progresses, then invade blood vessels and lymph nodes, spread from the primary organ along the lymph, deposit in the tissues of other organs, and then begin to proliferate. In terms of the present invention, the composition of the present invention can inhibit cancer metastasis by inhibiting some or all steps of cancer cells escaping from the primary organ, invading blood vessels and lymph nodes, and depositing in other organs, so the composition can be very efficiently applied to the purpose of inhibiting cancer metastasis. Preferably, the cancer metastasis described herein may refer to, but is not limited to, lung metastasis, more preferably lung metastasis of breast cancer, that is, the phenomenon that cancer cells growing in the primary organ breast are deposited and proliferated in another organ, the lung.

[0085] In the composition for inhibiting cancer metastasis of the present invention, the details of the compound represented by Formula 1 and the cancer are the same as those described in the above-mentioned composition for preventing, improving or treating cancer, so in order to avoid making this specification too complicated, their description will be omitted here.

[0086] The composition of the present invention may be in the form of a pharmaceutical composition, a food composition or a cosmetic composition.

[0087] In addition, the composition of the present invention can be further used in combination with other anticancer drugs. When the composition is used in combination with other anticancer drugs, it may show a significant effect of preventing or treating tumors by more effectively inhibiting tumor growth or metastasis.

[0088] The anti-tumor drug described herein may be, but is not limited to, at least one selected from the group consisting of nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, bosutinib, axitinib, cediranib, lestaurine, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, mistletoe, asparaginase, retinoic acid, hydroxyurea, dasatinib, estramonium, gemtuzumab ozogamicin, ibritumomab tiuxetan, heptadine, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, nitratechitosan), gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracil, oteracil, azacitidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, etorcitabine, flucitabine, capecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmopher, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vinblastine, idarubicin, mitomycin, bleomycin, actinomycin, pirarubicin , aclarubicin, peplomycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melparan, altretmine, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, folinic acid, tretinoin, exemestane, aminoglutethimide, anagrelide, olaparib, nabelbine, fadrozole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine, 5FU, vorinostat, entinostat, and carmustine.

[0089] The pharmaceutical composition of the present invention may be in the form of capsules, tablets, granules, injections, ointments, powders or drinks, and the pharmaceutical composition may be used for humans.

[0090] The pharmaceutical composition of the present invention can be formulated into oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, etc., and forms such as external preparations, suppositories, and sterile injection solutions according to conventional methods when used, but is not limited thereto. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. When used for oral administration, the pharmaceutically acceptable carrier may include a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a colorant, a flavoring agent, etc. When used for injection preparations, the pharmaceutically acceptable carrier may include a buffer, a preservative, an analgesic, a solubilizer, an isotonic agent, a stabilizer, etc. When used for topical administration, the pharmaceutically acceptable carrier may include a matrix, an excipient, a lubricant, a preservative, etc.

[0091] The pharmaceutical composition of the present invention can be formulated into various dosage forms together with the above-mentioned pharmaceutically acceptable carrier. For example, when used for oral administration, the pharmaceutical composition can be formulated into tablets, lozenges, capsules, elixirs, suspensions, syrups, thin films, etc. When used for injection, the pharmaceutical composition can be formulated into unit dose ampoules or multiple dose forms. In addition, the pharmaceutical composition can also be prepared into solutions, suspensions, tablets, capsules and sustained-release preparations.

[0092] In the present invention, the examples of carriers, excipients and diluents suitable for preparation include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxymethyl benzoate, propyl hydroxybenzoate, talcum powder, magnesium stearate and mineral oil. In addition, the pharmaceutical composition of the present invention can further include fillers, anticoagulants, lubricants, wetting agents, flavors, emulsifiers, preservatives, etc.

[0093] The administration route of the pharmaceutical composition of the present invention includes, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, percutaneous, subcutaneous, intraperitoneal, intranasal, gastrointestinal, topical, sublingual and rectal routes. Oral or parenteral administration is preferred. In the present invention, "parenteral administration" includes subcutaneous, percutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intradural, intralesional and intracranial injection or infusion techniques.

[0094] The pharmaceutical composition of the present invention may vary due to various factors, including the activity of the specific compound used, the patient's age, weight, general health, sex, diet, administration time, administration route, excretion rate, drug content and the severity of the specific disease to be prevented or treated. The dosage of the pharmaceutical composition can be appropriately selected by those skilled in the art according to the patient's condition, weight, severity of the disease, pharmaceutical dosage form and administration route and time, and can be 0.0001-50mg / kg / day or 0.001-50mg / kg / day. The pharmaceutical composition can be administered once or multiple times daily. The dosage is not intended to limit the scope of the present invention in any way. The pharmaceutical composition of the present invention can be formulated as a pill, a sugar-coated tablet, a capsule, a liquid, a gel, a syrup, a paste or a suspension.

[0095] The food composition of the present invention can be prepared into a variety of foods, for example, drinks, chewing gum, tea, vitamin complexes, powders, granules, tablets, capsules, candies, rice cakes, bread, etc.

[0096] When the compound of the present invention is used as an active ingredient of a food composition, it can be added in an amount of 0.1-50 wt % of the total weight of the food composition, but is not limited thereto.

[0097] When the food composition of the present invention is prepared into a drink, there is no particular limitation except that the drink contains the food composition in the percentage indicated. The drink may also contain various flavorings or natural carbohydrates, just like conventional beverages. Specifically, examples of the natural carbohydrates include monosaccharides (such as glucose), disaccharides (such as fructose), polysaccharides (such as sucrose), conventional sugars (such as dextrin, cyclodextrin, etc.), and sugar alcohols (such as xylitol, sorbitol, erythritol), etc. Examples of the flavorings include natural flavorings (sour wheat, stevia extract, such as rebaudioside A, glycyrrhizin, etc.) and artificial flavorings (saccharin, aspartame, etc.).

[0098] In addition, the food composition of the present invention may further contain various nutrients, vitamins, minerals (electrolytes), flavoring agents (such as synthetic flavoring agents and natural flavoring agents), coloring agents, pectin acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonizing agents used in carbonated beverages, etc.

[0099] The component comprised in food composition of the present invention can be used alone or in combination.Although the ratio of these additives is not critical to the present invention, it can be selected from, but not limited to the scope of 0.1 to about 50 weight parts in 100 weight parts of food composition of the present invention.

[0100] The cosmetic composition of the present invention can be prepared in the form of a skin softener, a nourishing liquid, a nourishing essence, a massage cream, a cosmetic bath water additive, a body liquid, a body milk, a bath oil, a baby oil, a baby powder, a shower gel, a shower milk, a bath cream, a sunscreen liquid, a sunscreen, a sunscreen, a skin liquid, a skin cream, an ultraviolet shielding cosmetic, a cleansing milk, a depilatory (for cosmetic purposes), a facial and body liquid, a facial and body cream, a skin whitening cream, a hand lotion, a hair lotion, a cosmetic cream, jasmine oil, a bath soap, a liquid soap, a cosmetic soap, a shampoo, a hand soap, a medicated soap (for non-medical purposes), a cream soap, a facial cleanser, a body cleanser, a scalp cleanser, a shampoo, a perfumed soap, a tooth whitening gel, a toothpaste, etc. For this purpose, the composition of the present invention may also contain a conventional solvent for preparing a cosmetic composition, or a suitable carrier, excipient or diluent.

[0101] In the present invention, the type of solvent that can be further added to the cosmetic composition is not particularly limited, and examples of the solvent may include water, saline, DMSO, or a combination thereof. In addition, examples of carriers, excipients, or diluents include, but are not limited to, purified water, oils, waxes, fatty acids, fatty acid alcohols, fatty acid esters, surfactants, humectants, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, lower alcohols, etc. In addition, if necessary, the cosmetic composition of the present invention may contain whitening agents, humectants, vitamins, ultraviolet light shielding agents, spices, dyes, antibiotics, antibacterial agents, and antifungal agents.

[0102] In the present invention, examples of oils include hydrogenated vegetable oils, castor oil, cottonseed oil, olive oil, palm kernel oil, jojoba oil, and avocado oil, and examples of waxes include beeswax, refined wax, carnauba wax, candelilla wax, montan wax, ozokerite, liquid paraffin, and lanolin.

[0103] In the present invention, examples of fatty acids include stearic acid, linoleic acid, linolenic acid and oleic acid; examples of fatty acid alcohols include cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol and cetyl alcohol; examples of fatty acid esters include isopropyl myristate, isopropyl palmitate and butyl stearate. Examples of surfactants include cationic surfactants, anionic surfactants and nonionic surfactants, all of which are known in the art. Among these surfactants, surfactants derived from natural products are preferred if possible.

[0104] In addition to the above ingredients, the cosmetic composition of the present invention may further contain a moisturizer, a thickener, an antioxidant, etc., which are well known in the cosmetic field, and the types and amounts thereof are known in the art.

[0105] Another embodiment of the present invention provides a method for preventing or treating cancer, the method comprising the steps of administering an effective amount of a compound represented by the following formula 1 to a subject in need thereof:

[0106] [Formula 1]

[0107]

[0108] wherein n, m, X and Y are respectively as defined in the above formula 1 of the composition for preventing, ameliorating or treating cancer.

[0109] In the method for preventing or treating cancer of the present invention, the details of the compound shown in Formula 1 and the cancer are the same as those described in the composition for preventing, improving or treating cancer. In order to avoid making this specification too complicated, their description will be omitted here.

[0110] Another embodiment of the present invention provides a method for inhibiting cancer metastasis, the method comprising the step of administering an effective amount of a compound represented by the following formula I to a subject in need thereof:

[0111] [Formula 1]

[0112]

[0113] wherein n, m, X and Y are respectively as defined in the above formula 1 of the composition for preventing, ameliorating or treating cancer.

[0114] In the method for inhibiting cancer metastasis of the present invention, the details of the compound represented by Formula 1 and the cancer are the same as those described in the composition for inhibiting cancer metastasis, and thus their description will be omitted here to avoid making this specification too complicated.

[0115] As used herein, the term "administering" refers to providing a given compound of the invention to a subject in any appropriate manner.

[0116] In the present invention, the term "subject" in need of medication may include mammals and non-mammals. Here, examples of the mammals include, but are not limited to, humans; non-human primates, such as chimpanzees, or other apes or monkeys; farm animals, such as cattle, horses, sheep, goats, pigs; livestock, such as rabbits, dogs and cats; experimental animals, including rodents, such as rats, mice or guinea pigs. In addition, in the present invention, examples of the non-mammals include, but are not limited to, birds or fish.

[0117] In the present invention, the preparation form of the compound administered as described above is not particularly limited, and the composition can be administered as a solid preparation, a liquid preparation or an aerosol preparation for inhalation, and can be used as a solid form preparation for rapid conversion into a liquid form preparation before oral or parenteral administration. The compound of the present invention can be formulated and administered as an oral dosage form such as powder, granules, capsules, tablets, aqueous suspensions, etc., as well as forms such as external preparations, suppositories, and sterile injection solutions, but are not limited thereto.

[0118] In addition, in the present invention, pharmaceutically acceptable carriers can be additionally used together with the compounds of the present invention. Here, when used for oral administration, the pharmaceutically acceptable carrier may include adhesives, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavoring agents, etc. When used for injection, the pharmaceutically acceptable carrier may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc. When used for topical administration, the pharmaceutically acceptable carrier may include matrix, excipients, lubricants, preservatives, etc. The preparation of the compound of the present invention can be prepared in a variety of ways by mixing with the above-mentioned pharmaceutically acceptable carriers. For example, when used for oral administration, the compound can be formulated into tablets, lozenges, capsules, elixirs, suspensions, syrups, film tablets, etc. When used for injection, the compound can be formulated into unit dose ampoules or multiple dose forms. In addition, the compound can also be prepared as solutions, suspensions, tablets, capsules and sustained-release preparations.

[0119] Wherein, the example of carrier, excipient and diluent suitable for preparation comprises lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxymethyl benzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.In addition, described composition can further comprise filler, anticoagulant, lubricant, wetting agent, essence, emulsifier, preservative etc.

[0120] The administration routes of the compounds of the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, gastrointestinal, topical, sublingual and rectal routes. Oral or parenteral administration is preferred.

[0121] In the present invention, "parenteral administration" includes subcutaneous, percutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intradural, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention can also be prepared as a suppository for rectal administration.

[0122] In the present invention, the term "pharmaceutically effective amount" refers to a sufficient drug dosage that can provide a desired biological result. The result can be to reduce and / or alleviate the signs, symptoms or causes of the disease, or any other desired changes in the biological system. For example, an "effective amount" for treatment is the amount of the compound of the present invention required to provide a clinically significant reduction in the disease. In any individual case, an appropriate "effective" amount can be determined by a person of ordinary skill in the art through routine experiments. Therefore, an "effective amount" generally refers to the amount of an active substance with a therapeutic effect. In the present invention, the active substance is a cancer cell growth inhibitor, a cancer prevention, improvement, or therapeutic agent, and a cancer metastasis inhibitor.

[0123] The dosage of the compound of the present invention may vary due to various factors, including the activity of the specific compound used, the patient's age, body weight, general health, sex, diet, administration time, route of administration, excretion rate, drug content and the severity of the specific disease to be prevented or treated. The dosage of the compound can be appropriately selected by those skilled in the art according to the patient's condition, body weight, severity of the disease, pharmaceutical dosage form and route of administration and time, and can be 0.0001-100mg / kg / day or 0.001-100mg / kg / day. The compound can be administered once or multiple times daily. The dosage is not intended to limit the scope of the present invention in any way. The compound of the present invention can be formulated as a pill, a sugar-coated tablet, a capsule, a liquid, a gel, a syrup, a paste or a suspension.

[0124] The compounds of the present invention can be used alone or in combination with surgery, radiotherapy, hormone therapy, chemotherapy and biological response modifiers.

[0125] In addition, the compounds of the present invention can be further used in combination with other anticancer drugs. Wherein, the antitumor drug can be, but is not limited to, at least one selected from the group consisting of nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, bosutinib, axitinib, cediranib, lestaurine, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, mistletoe, asparaginase, retinoic acid, hydroxyurea, dasatinib, estramonin, gemtuzumab ozogamicin, ibritumomab tiuxetan, heptadine, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan (holmium nitrate chitosan) chitosan), gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracil, oteracil, azacitidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, etorcitabine, flucitabine, capecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmopher, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vinblastine, idarubicin, mitomycin, bleomycin, actinomycin, pirarubicin, aclarubicin alanine, pelocycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melparan, altretmine, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, folinic acid, tretinoin, exemestane, aminoglutethimide, anagrelide, olaparib, nabelbine, fadrozole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine, 5FU, vorinostat, entinostat, and carmustine.

[0126] Beneficial Effects

[0127] The composition of the present invention can not only be used to prevent, improve, or treat cancer very efficiently, but also can inhibit cancer metastasis by inhibiting the growth of cancer cells and very efficiently inhibiting the metastasis of cancer cells to other tissues. BRIEF DESCRIPTION OF THE DRAWINGS

[0129] Figure 1 Shown are the results of evaluating the increase in AMPK protein stability by Western blot experiments in one example of the present invention.

[0130] Figure 2a and 2bShown are the results of evaluating the effect of inhibiting the survival and growth of cancer cell lines by cell viability assay (a) and clonogenic assay (b) in one example of the present invention.

[0131] Figure 3a Shown are the results of evaluating the effect of inhibiting the survival and growth of cancer cell lines by cell viability assay in one embodiment of the present invention.

[0132] Figure 3b Shown are the results of evaluating the AMPK activation ability in cancer cells by p-AMPK (Thr172) expression level in one example of the present invention.

[0133] Figure 4a-4c The results of evaluating the survival rate inhibitory effect and AMPK agonistic ability of AMPK activators, the compound of Preparation Example 6 and the compound of Preparation Example 8 having a similar structure, against gastric cancer, brain cancer and pancreatic cancer cell lines in one embodiment of the present invention are shown.

[0134] Figure 5 Shown are the results of evaluating the effect of inducing apoptosis in cancer cell lines using flow cytometry assay in one example of the present invention.

[0135] Figure 6 Shown are the results of evaluating the effect of inducing apoptosis in cancer cell lines using Western blot experiments in one example of the present invention.

[0136] Figure 7a and 7b Shown are the results of evaluating the effect of inhibiting EMI of cancer cell lines using Western blot experiments in one example of the present invention.

[0137] Figures 8a-8d Shown are the results of evaluating the tumor regression effect in an animal model in one embodiment of the present invention.

[0138] Figure 9a and 9b Shown are the results of evaluating the tumor metastasis inhibition effect in an animal model in one embodiment of the present invention.

[0139] The best way

[0140] One embodiment of the present invention relates to a composition for preventing, improving, or treating cancer, wherein the composition contains a compound represented by the following formula 1 as an active ingredient:

[0141] Another embodiment of the present invention relates to a composition for inhibiting cancer metastasis, wherein the composition contains a compound represented by the following formula 1 as an active ingredient:

[0142] Another embodiment of the present invention is directed to a method for preventing or treating cancer, the method comprising the step of administering an effective amount of a compound represented by the following formula 1 to a subject in need thereof:

[0143] Another embodiment of the present invention is directed to a method for inhibiting cancer metastasis, the method comprising the step of administering an effective amount of a compound represented by the following formula I to a subject in need thereof:

[0144] [Formula 1]

[0145]

[0146] in:

[0147] L1 and L2 are each independently selected from the following group: C3-C 40 Cycloalkylene, C6-C 60 Arylene, and heteroarylene having 5 to 60 nuclear atoms;

[0148] X and Y are each independently selected from the group consisting of deuterium, halogen, cyano, nitro, sulfonyl, C1-C 10 Alkylsulfonyl, azide group, hydroxyl, C1-C 40 Alkyl, C2-C 40 Alkenyl, C1-C 40 Alkoxy, unsubstituted or substituted C6-C 60 Aryloxy, unsubstituted or substituted C3-C 40 Cycloalkyl, unsubstituted or substituted heterocycloalkyl having 3 to 20 nuclear atoms, unsubstituted or substituted C6-C 60 Aryl, unsubstituted or substituted heteroaryl having 5 to 60 nuclear atoms, and -NR'R";

[0149] R' and R" are each independently selected from the group consisting of hydrogen, C1-C 10 Alkyl, C6-C 60 Aryl, C3-C 40 Cycloalkyl, C6-C 60 Arylsulfonyl, and heteroaryl having 5 to 60 nuclear atoms;

[0150] n and m are each independently an integer from 0 to 5, provided that n and m cannot be 0 at the same time;

[0151] When there are multiple X or Y, the multiple X or Y may be the same as or different from each other; and

[0152] The arylsulfonyl group in each of R' and R" may be unsubstituted or substituted with at least one substituent selected from the group consisting of deuterium, halogen, and nitro.

[0153] Embodiments of the invention

[0154] Below, the present invention will be described in more detail by combining embodiment.Obviously, to those skilled in the art, these embodiments are only used to explain the present invention in more detail, and according to the subject matter of the present invention, the scope of the present invention is not limited by these embodiments. Example

[0155] [Preparation Example 1-15] Synthesis of candidate compounds

[0156] The compounds of Preparation Examples 1 to 15 shown in Table 1 below were prepared using Method 1 or Method 2.

[0157] [Table 1]

[0158]

[0159]

[0160] [Method 1]

[0161] The method comprises the steps of: adding an acetophenone derivative (1 equivalent), a benzaldehyde derivative (1 equivalent) and NaOH (1 equivalent) into an ethanol solvent, and then stirring at room temperature; after the reaction is completed, adding water to the reaction mixture, and then extracting with ethyl acetate; and collecting an organic solvent layer, washing the collected material once with water, drying the washed material with anhydrous MgSO4, removing the solvent by distillation under reduced pressure, and purifying the residue by silica gel chromatography.

[0162] [Method 2]

[0163] The method comprises the steps of: adding an acetophenone derivative (1 equivalent), a 4-(tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde derivative (1 equivalent) and NaOH (1 equivalent) in an ethanol solvent, and then stirring at room temperature; after the reaction is completed, adding 4M HCl to the reaction mixture, and then stirring for 20 minutes, adding water, and then extracting with ethyl acetate; and collecting an organic solvent layer, washing the collected material once with water, drying the washed material with anhydrous MgSO4, removing the solvent by distillation under reduced pressure, and purifying the residue by silica gel chromatography.

[0164] [Example 1] (E)-1-(4-aminophenyl)-3-(2,4-dimethoxyphenyl)prop-2-en-1-one (YE-01)

[0165] According to the above method 1, 4-aminoacetophenone (0.30 g, 2.22 mmol), 2,4-dimethoxybenzaldehyde (0.37 g, 2.22 mmol) and NaOH (0.09 g, 2.22 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:2 → 1:1) to obtain the compound of Preparation Example 1 (0.15 g, yield 23.0%) as a yellow solid. f0.33 (ethyl acetate / n-hexane=1:1); 1 H-NMR (400MHz, CDCl3) δ3.85(s,3H),3.89(s,3H),6.47(d,J=2.0Hz,1H),6.52(dd,J=8.4,2.4Hz,1H),6.69(d ,J=8.4Hz,2H),7.55(d,J=15.6Hz,1H),7.56(d,J=8.8Hz,1H),7.92(d,J=8.4Hz,2H),8.02(d,J=15.6Hz,1H); 13 C-NMR (100MHz, CDCl3)55.6,55.7,98.7,105.5,114.1,117.8,120.7,129.4,130.8,131.1,139.0,150.9,160.4,162.8,189.1ppm.

[0166] [Example 2] (E)-1-(4-aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one (YE-02)

[0167] According to the above method 1, 4-aminoacetophenone (0.50 g, 3.70 mmol), 2,5-dimethoxybenzaldehyde (0.62 g, 3.70 mmol) and NaOH (0.15 g, 3.70 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:2 → 1:1) to obtain the compound of Preparation Example 2 (0.66 g, yield 62.5%) as a yellow solid. f 0.36 (ethyl acetate / n-hexane=1:1); 1 H-NMR(400MHz, CDCl3)δ3.81(s,3H),3.85(s,3H),4.19(br s,2H),6.69(d,J=8.8Hz,2H),6.86(d,J=8.8Hz,1H),6.91(dd,J=8.8,2.8Hz,1H),7.16( d,J=2.8Hz,1H),7.58(d,J=15.6Hz,1H),7.92(d,J=8.8Hz,2H),8.04(d,J=15.6Hz,1H); 13 C-NMR (100MHz, CDCl3)56.0,56.3,112.7,113.9,114.1,116.8,123.3,125.2,128.9,131.3,138.6,151.2,153.4,153.7,188.8ppm.

[0168] [Example 3] (E)-1-(3-aminophenyl)-3-(4-methoxyphenyl)prop-2-en-1-one (YE-03)

[0169] According to the above method 1, 3-aminoacetophenone (1.00 g, 7.40 mmol), 4-methoxybenzaldehyde (1.00 g, 7.40 mmol) and NaOH (0.30 g, 7.40 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:2 → 1:1) to obtain the compound of Preparation Example 3 (0.83 g, yield 62.5%) as an orange solid. f 0.40 (ethyl acetate / n-hexane=1:1); 1 H-NMR(400MHz,CDCl3)δ3.83(br s,2H),3.85(s,3H),6.88(ddd,J=8.0,2.4,0.8Hz,1H),6.93(d,J=8.8Hz,2H),7.27(dd,J=8.0,7.6Hz,1H),7.31(dd,J=2 .0,1.6Hz,1H),7.36(d,J=15.6Hz,1H),7.38(ddd,J=7.6,1.6,0.8Hz,1H),7.59(d,J=8.8Hz,2H),7.76(d,J=15.6Hz,1H); 13 C-NMR (100MHz, CDCl3)56.0,56.3,112.7,113.9,114.1,116.8,123.3,125.2,128.9,131.3,138.6,151.2,153.4,153.7,188.8ppm.

[0170] [Example 4] (E)-1-(3-aminophenyl)-3-(4-hydroxy-2-methoxyphenyl)prop-2-en-1-one (YE- 04)

[0171] According to the above method 2, 3-aminoacetophenone (0.40 g, 2.96 mmol), 2-methoxy-4-((tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.70 g, 2.96 mmol) and NaOH (0.12 g, 2.96 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:1) to obtain the compound of Preparation Example 4 (0.28 g, yield 35.1%) as an orange solid. f 0.25 (ethyl acetate / n-hexane=1:1); 1H-NMR (400MHz, CDCl3) δ3.71(s,3H),6.30(d,J=2.4Hz,1H),6.34(dd,J=8.4,2.0Hz,1H),6.72(ddd,J=8.0,2.4,0.8Hz,1H),7.08(dd,J=8.4,8.0 Hz,1H),7.13(d,J=2.4Hz,1H),7.16(ddd,J=7.6,7.6,0.8Hz,1H),7.29(d,J=15.6Hz,1H),7.32(d,J=8.4Hz,1H),7.85(d,J=15.6Hz,1H),9.35(br s,1H); 13 C-NMR (100MHz, CDCl3)55.3,99.1,108.2,114.1,115.3,118.1,118.7,119.3,129.1,130.6,139.8,140.3,147.0,60.5,161.4,191.1ppm.

[0172] [Example 5] (E)-1-(3-aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one (YE-05)

[0173] According to the above method 1, 4-aminoacetophenone (0.50 g, 3.70 mmol), 2,5-dimethoxybenzaldehyde (0.62 g, 3.70 mmol) and NaOH (0.15 g, 3.70 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:3) to obtain the compound of Preparation Example 5 (0.27 g, yield 25.4%) as a yellow solid. f 0.62 (ethyl acetate / n-hexane=1:1); 1 H-NMR (400MHz, CDCl3) δ3.81(s,3H),3.86(s,3H),6.87(d,J=8.8Hz,1H),6.88(ddd,J=8.4,2.0,0.8Hz,1H),6.94(dd,J=8.8,2.8Hz,1H),7.16(d,J=2. 8Hz,1H),7.27(dd,J=8.0,8.0Hz,1H),7.31(dd,J=2.8,2.8Hz,1H),7.38(d dd,J=8.0,2.0,1.6Hz,1H),7.53(d,J=15.6Hz,1H),8.06(d,J=15.6Hz,1H); 13C-NMR (100MHz, CDCl3)56.1,56.3,112.7,113.9,114.7,117.4,119.1,119.4,123.6,124.8,129.6,139.8,140.0,147.0,153.5,153.7,191.4ppm.

[0174] [Example 6] (E)-4-chloro-N-(4-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide (YE-06)

[0175] 4-Chlorobenzenesulfonyl chloride (0.75 g, 3.54 mmol) was added to a CH2Cl2 solution of the compound of Preparation Example 2 (0.67 g, 2.36 mmol) and triethylamine (TEA, 0.26 g, 2.60 mmol), and the mixture was stirred at room temperature for 24 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic solvent layer was collected, washed with saturated NaHCO3, and dried over anhydrous MgSO4. The solvent was then distilled off under reduced pressure, and the residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:2) to obtain the compound of Preparation Example 6 (0.55 g, yield 50.9%) as a yellow solid. f 0.18 (ethyl acetate / n-hexane=1:2); 1 H-NMR (400MHz, DMSO-d6) δ3.74(s,3H),3.80(s,3H),6.84(d,J=8.8Hz,1H),6.88(dd,J=8.8,2.4Hz,1H),7.16(d,J=2.4Hz,1H),7.20(d,J=8. 4Hz, 2H), 7.40 (d, J = 8.4Hz, 2H), 7.54 (d, J = 15.6Hz, 1H), 7.74 (d, J = 8.4Hz, 2H), 7.85 (d, J = 8.4Hz, 2H), 7.91 (d, J = 15.6Hz, 1H), 10.58 (s, 1H); 13 C-NMR(100MHz,DMSO-d6)55.2,55.6,112.1,112.7,117.0,118.0,121.6,123.5,1 28.0,128.1,128.7,129.4,132.9,137.8,138.3,141.5,152.5,152.9,187.9ppm.

[0176] [Example 7] (E)-4-chloro-N-(3-(3-(4-methoxyphenyl)acryloyl)phenyl)benzenesulfonamide (YE-07)

[0177] N-(3-acetylphenyl)-4-chlorobenzenesulfonamide (0.10 g, 0.32 mmol), 4-methoxybenzaldehyde (0.04 g, 0.32 mmol) and NaOH (0.03 g, 0.80 mmol) were added to the ethanol solvent, followed by stirring at room temperature for 72 hours. A dilute aqueous hydrochloric acid solution was added to the reaction mixture, followed by extraction with ethyl acetate. The organic solvent layer was collected, washed with water, and dried over anhydrous MgSO4. The solvent was removed by distillation under reduced pressure, and the residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:3→1:1) to obtain the compound of Preparation Example 7 (0.01 g, yield 6.5%) as a yellow solid. 1 H-NMR (400MHz, CDCl3) δ3.84(s,3H),6.92(d,J=8.8Hz,2H),7.28(d,J=15.6Hz,1H),7.33(dd,J=8.0,7.6Hz,1H),7.36(d,J=8.8Hz ,2H),7.41(ddd,J=8.0,2.0,1.2Hz,1H),7.57(d,J=8.8Hz,2H),7.68(ddd,J=7.6,1.6,1.2Hz,1H),7.71-7.78(m,4H),9.25(s,1H); 13 C-NMR (100MHz, CDCl3)55.6,114.6,119.5,121.1,124.9,125.0,127.6,128.8,129.4,129.7,130.5,137.

[0178] [Example 8] (E)-4-Chloro-N-(3-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide (YE-08)

[0179] 4-Chlorobenzenesulfonyl chloride (0.09 g, 0.43 mmol) was added to a CH2Cl2 solution of the compound of Example 5 (0.12 g, 0.43 mmol) and triethylamine (0.03 g, 2.60 mmol), and the mixture was stirred at room temperature for 24 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic solvent layer was collected, washed with saturated NaHCO3, and dried over anhydrous MgSO4. The solvent was then removed by distillation under reduced pressure, and the residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:3) to obtain the compound of Example 8 (0.08 g, yield 37.3%) as a yellow solid. f 0.33 (ethyl acetate / n-hexane=1:1); 1H-NMR (400MHz, CDCl3) δ3.82(s,3H),3.88(s,3H),6.88(d,J=8.8Hz,1H),6.96(dd,J=8.4,2.8Hz,1H),7.15(d,J=2.8Hz,1H),7.40(d,J=8. 8Hz,2H),7.41-7.43(m,2H),7.55(d,J=16.0Hz,1H),7.73(d,J=8.8Hz,2H),7.71-7.72(m,1H),7.76-7.79(m,1H),8.08(d,J=16.0Hz,1H); 13 C-NMR (100MHz, CDCl3)56.1,56.3,112.7,114.5,117.9,121.6,122.8,124.4,125.5,12 5.8,128.9,129.7,130.0,137.1,137.7,139.9,140.0,141.6,153.7,153.8,190.3ppm.

[0180] [Example 9] (E)-4-chloro-N-((4-chlorophenyl)sulfonyl)-N-(3-(3-(2,5-dimethoxyphenyl)propene Acyl)phenyl)benzenesulfonamide (YE-09)

[0181] 4-Chlorobenzenesulfonyl chloride (0.25 g, 1.17 mmol) was added to a CH2Cl2 solution of the compound of Example 5 (0.22 g, 0.78 mmol) and trimethylamine (0.22 g, 2.12 mmol), and the mixture was stirred at room temperature for 24 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic solvent layer was collected, washed with saturated NaHCO3, and dried over anhydrous MgSO4. The solvent was then removed by distillation under reduced pressure, and the residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:4) to obtain the compound of Example 8 (0.20 g, yield 39.8%) as a yellow solid. f 0.77 (ethyl acetate / n-hexane=1:1); 1H-NMR (400MHz, CDCl3) δ3.83 (s, 3H), 3.86 (s, 3H), 6.89 (d, J = 9.2Hz, 1H), 6.97 (d d,J=9.2,3.2Hz,1H),7.13(d,J=2.8Hz,1H),7.19(dd,J=8.4,2.0Hz,1H),7.42(d ,J=16.0Hz,1H),7.52(d,J=8.4Hz,1H),7.54(d,J=8.8Hz,4H),7.66(dd,J=1.6,1 .6Hz,1H),7.89(d,J=8.8Hz,4H),8.05(d,J=16.0Hz,1H),8.09(d,J=8.0Hz,1H),; 13 C-NMR (100MHz, CDCl3)56.1,56.3,112.7,114.3,117.9,122.9,124.3,129.8,129.9,13 0.3,130.6,131.6,134.6,135.2,137.8,140.2,141.4,141.8,153.7,153.8,189.9ppm.

[0182] [Example 10] (E)-1-(4-aminophenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one (YE-10)

[0183] According to the above method 1, 4-aminoacetophenone (0.40 g, 2.96 mmol), 4-(piperidin-1-yl)benzaldehyde (0.56 g, 2.96 mmol) and NaOH (0.12 g, 2.96 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: MeOH:CHCl3=1:19) to obtain the compound of Preparation Example 10 (0.28 g, yield 30.9%) as an orange solid. f 00.43 (ethyl acetate / n-hexane=1:1); 1 H-NMR (400MHz, CDCl3) δ1.62-1.71 (m, 6H), 3.29 (t, J = 5.2Hz, 4H), 4.10 (br s, 2H), 6.69 (d, J = 8.8Hz, 2H), 6.89 (d, J = 8.8Hz, 2H), 7.37 (d, J = 15.6Hz, 1H), 7.53 (d, J = 8.8Hz, 2H), 7.75 (d, J = 15.6Hz, 1H), 7.95 (d, J = 8.8Hz, 2H).

[0184] [Example 11] (E)-1-(3-aminophenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one (YE-11)

[0185] According to the above method 1, 3-aminoacetophenone (0.40 g, 2.96 mmol), 4-(piperidin-1-yl)benzaldehyde (0.56 g, 2.96 mmol) and NaOH (0.12 g, 2.96 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: MeOH:CHCl3=1:19) to obtain the compound of Preparation Example 11 (0.45 g, yield 49.6%) as an orange solid. f 0.47 (ethyl acetate / n-hexane=1:1); 1 H-NMR (400MHz, CDCl3) δ1.61-1.72 (m, 6H), 3.31 (t, J = 5.6Hz, 4H), 3.80 (br s,2H),6.86(ddd,J=8.0,2.4,0.8Hz,1H),6.89(d,J=8.8Hz,2H),7.28(dd,J=8.0,8.0Hz,1H),7.30(d,J=15.6Hz,1 H),7.31(dd,J=2.0,2.0Hz,1H),7.37(ddd,J=8.8,1.2,1.2Hz,1H),7.53(d,J=8.8Hz,2H),7.75(d,J=15.6Hz,1H); 13 C-NMR (100MHz, CDCl3) 24.6, 25.7, 49.3, 114.7, 115.0, 118.4, 119.0, 119.1, 124.7, 129.5, 130.4, 140.3, 145.4, 146.9, 153.4, 191.1ppm.

[0186] [Example 12] (E)-1-(4-hydroxyphenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one (YE-12)

[0187] According to the above method 2, 1-4-(tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethane-1-one (0.50 g, 2.27 mmol), 4-(piperidin-1-yl)benzaldehyde (0.43 g, 2.27 mmol) and NaOH (0.09 g, 2.27 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:3→1:1) to obtain the compound of Preparation Example 12 (0.24 g, yield 33.7%) as an orange solid. f 0.17 (ethyl acetate / n-hexane=1:1); 1H-NMR (400MHz, CDCl3) δ1.58-1.65(m,6H),3.25(t,J=5.6Hz,4H),6.84(d,J=8.8Hz,2H),6.87(d,J=8.8Hz,2H ),7.32(d,J=15.6Hz,1H),7.48(d,J=8.8Hz,2H),7.68(d,J=15.6Hz,1H),7.90(d,J=8.8Hz,2H),9.34(s,1H); 13 C-NMR (100MHz, CDCl3) 24.4, 25.5, 49.2, 114.9, 115.6, 117.8, 124.8, 130.1, 130.6, 130.9, 144.2, 153.1, 161.7, 189.0ppm.

[0188] [Example 13] (E)-1-(2,4-dihydroxyphenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one (YE-13)

[0189] According to the above method 2, 1-4-(tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethane-1-one (0.50 g, 2.12 mmol), 4-(piperidin-1-yl)benzaldehyde (0.40 g, 2.12 mmol) and Ba(OH)2·8H2O (0.73 g, 2.33 mmol) were used as raw materials. The residue was purified by silica gel chromatography (developing solvent: ethyl acetate / n-hexane = 1:1) to obtain the compound of Preparation Example 13 (0.12 g, yield 16.8%) as an orange solid. f 0.66 (ethyl acetate / n-hexane=1:1); 1 H-NMR (400MHz, CDCl3) δ1.58-1.65(m,6H),3.27(t,J=5.6Hz,4H),6.37-6.40(m,2H),6.84(d,J=8.8Hz,2H),7.34( d,J=15.2Hz,1H),7.49(d,J=8.8Hz,2H),7.73(d,J=8.4Hz,1H),7.76(d,J=15.2Hz,1H),9.64(s,1H),13.6(s,1H).

[0190] [Example 14] (E)-3-(4-hydroxy-2-methoxyphenyl)-1-(4-(piperazin-1-yl)phenyl)prop-2-ene-1- Ketone (YE-14)

[0191] According to the above method 2, 1-4-(tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethane-1-one (0.50 g, 2.45 mmol), 2-methoxy-4-(tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.58 g, 2.45 mmol) and NaOH (0.20 g, 4.90 mmol) were used as raw materials. After removing the solvent, the obtained solid was treated with a mixed solvent of ethyl acetate / n-hexane. The obtained solid was filtered and dried in vacuo to obtain the compound of Production Example 14 (0.28 g, yield 33.8%) as an orange solid. 1 H-NMR (400MHz, DMSO-d6) δ2.89(dd,J=7.2,3.2Hz,4H),3.26(dd,J=7.2,4.0Hz,4H),3.82(s,3H),6.40(dd,J=8.8,2.0Hz,1H),6.41( d,J=1.6Hz,1H),6.89(d,J=8.8Hz,2H),7.51(d,J=15.6Hz,1H),7.55(d,J=8.8Hz,1H),7.87(d,J=15.6Hz,1H),7.90(d,J=8.8Hz,2H); 13 C-NMR (100MHz, DMSO-d6) 45.1, 47.5, 55.1, 98.7, 107.9, 112.9, 114.7, 118.0, 127.7, 129.7, 129.8, 137.7, 153.7, 159.7, 161.2, 186.8ppm.

[0192] [Example 15] (E)-3-(4-hydroxyphenyl)-1-(4-(methylpiperazin-1-yl)phenyl)prop-2-en-1-one (YE-15)

[0193] According to the above method 2, 1-(4-(tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethane-1-one (0.50 g, 2.29 mmol), 4-(tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.47 g, 2.29 mmol) and NaOH (0.09 g, 2.29 mmol) were used as raw materials. After removing the solvent, the obtained solid was treated with a mixed solvent of ethyl acetate / n-hexane. The obtained solid was filtered and dried in vacuo to obtain the compound of Production Example 15 (0.70 g, yield 94.8%) as an orange solid. 1H-NMR(400MHz,,CDCl3)δ2.33(s,3H),2.55(t,J=5.2Hz,4H),3.37(t,J=5.2Hz,4H),6.86(d,J=8.8Hz,2H),6.89 (d,J=8.8Hz,2H),7.38(d,J=15.6Hz,1H),7.49(d,J=8.8Hz,2H),7.72(d,J=15.6Hz,1H),7.95(d,J=8.8Hz,2H); 13 C-NMR (100MHz, CDCl3) 46.2, 47.4, 54.8, 113.7, 116.2, 118.8, 126.8, 128.7, 130.3, 130.6, 143.7, 154.0, 159.7, 188.4ppm.

[0194] [Experimental methods]

[0195] [Experimental method 1] Test cell lines and cell line culture methods

[0196] The cytotoxicity of the compound of Preparation Example 6 of the present invention was evaluated using human cell lines (VERO, HFL-1, L929, NIH 3T3, and CHO-K1). In addition, the following tumor cell lines were used to evaluate the ability of the compound of Preparation Example 6 and 13 compounds of the present invention (including structurally similar compounds (YE-01, 02, 03, 04, 05, 08, 09, 10, 11, 12, 13, 14 and 15)) to inhibit the growth of tumor cell lines: human gastric cancer cell lines (NCI-N87, and SNU-216), human brain cancer cell lines (U-87MG, and Hs 683), human pancreatic cell lines (AsPC-1, and MIA PaCa-2), human breast cancer cell lines (BT549, MDA-MB-231 and Hs578t) and mouse breast cancer cell line 4T1 or 4T1-luc (a cell line constructed so that luciferase can be expressed in the 4T1 cell line and the location of the cells can be visually confirmed by simple pretreatment).

[0197] Each cell line is provided by ATCC (American Type Culture Collection), JCRB Cell Bank (Japan Research Bioresource Cell Bank) or Korean Cell Line Bank (KCRB). Each cell line is cultured according to the culture medium and culture conditions recorded on the ATCC official website. In addition, to avoid mycoplasma infection that may affect gene expression in cell lines, routine checks for mycoplasma infection are performed during cell line culture. When mycoplasma infection is confirmed, anti-mycoplasma antibiotics are used for 1-2 weeks. Here, the 4T1-luc cell line is used to construct a xenogeneic / orthotopic animal model.

[0198] [Experimental method 2] Conditions for treating cell lines with the compound of Preparation Example 6

[0199] The compound of Preparation Example 6 was dissolved in DMSO at a concentration of 1-50 μM. Subsequently, the concentration and time at which the compound of Preparation Example 6 showed the best survival rate and a clear trend for each breast cancer cell line of Experimental Method 1 were detected. At the same time, the control group of Preparation Example 6 compound therapy was treated with DMSO only. In order to avoid causing interference such as inhibiting cell survival rate, DMSO did not exceed 5% of the total amount of the culture medium during DMSO therapy.

[0200] [Experimental method 3] Methods for measuring stabilization and activation of AMPK (AMP-activated protein kinase)

[0201] Cellular thermal shift analysis (CTSA) was performed to detect whether the compound of Preparation Example 6 and 13 compounds (including structurally similar compounds (YE-01, 02, 03, 04, 05, 08, 09, 10, 11, 12, 13, 14 and 15) of Preparation Examples 1-5 and 8-15) bind to the AMP binding site on AMPK. Specifically, trypsin was used to separate cultured human breast cancer-derived cells (BT549 or MDA-MB-231 cell lines) from a culture dish. The separated cell lines were diluted in PBS, and an equal amount of the cell lines were allocated to each test tube for PCR and heated from room temperature to 65°C for 3 minutes. After heating, liquid nitrogen was used to separate proteins from the cell lines, and a Western blotting experiment was performed to confirm the temperature at which AMPK undergoes high temperature denaturation.

[0202] [3-1] The breast cancer cell line of the above experimental method 1 was treated with the compound of Preparation Example 6 at a concentration of 0-30 μM, followed by culturing for 24 hours. Afterwards, the cells were collected and lysed in RIPA buffer, protein electrophoresis was performed using SDS-PAGE, and then protein expression levels were analyzed using specific anti-AMPK and phospho-AMPK (Thr172) antibodies. GAPDH or β-actin was used as an internal reference for protein quantification.

[0203] [3-2] The three breast cancer cell lines (4T1, BT549, and MDA-MB-231) of Experimental Method 1 were treated with the compound of Preparation Example 6 and 13 compounds (including compounds with similar structures (YE-01, 02, 03, 04, 05, 08, 09, 10, 11, 12, 13, 14, and 15)) at a concentration of 10 μM, respectively, and then cultured for 24 hours. In addition, two types of gastric cancer cell lines, brain cancer cell lines, and pancreatic cancer cell lines were treated with the compound of Preparation Example 6 or the compound with similar structure of Preparation Example 8 at a concentration of 10 μM, respectively, and then cultured for 12 hours. Afterwards, the cells were collected and lysed in RIPA buffer, protein electrophoresis was performed using SDS-PAGE, and then the protein expression level was analyzed using antibodies specific for AMPK and phospho-AMPK (Thr172). GAPDH or β-actin was used as an internal reference control for protein quantification.

[0204] [Experimental method 4] Methods for determining growth inhibition and apoptosis levels in tumor cell lines

[0205] [4-1] Cell viability assay

[0206] [4-1-1] The breast cancer cell line of Experimental Method 1 was treated with 5-aminoimidazole-4-carboxamide ribonucleotide (not the compound of Preparation Example 6) at a concentration of 0-30 μM or the compound of Preparation Example 6, followed by culturing for 24 hours, 48 ​​hours and 72 hours. Thereafter, the cultured cells were treated with the WST-1 reagent, and the absorbance at a wavelength of 450 nm was measured. Here, the WST-1 reagent can determine the cell survival rate by changing the color of the culture medium according to the level of mitochondrial dehydrogenase present in the cells.

[0207] [4-1-2] The breast cancer cell lines (4T1, BT549, or MDA-MB-231) of Experimental Method 1 were treated with the compound of Preparation Example 6 and 13 compounds (including compounds with similar structures (YE-01, 02, 03, 04, 05, 08, 09, 10, 11, 12, 13, 14, and 15)) at a concentration of 0-50 μM, respectively, and then cultured for 24 hours. Three types of gastric cancer cell lines, brain cancer cell lines, and pancreatic cancer cell lines were treated with the compound of Preparation Example 6 or the compound with similar structure of Preparation Example 8 at a concentration of 0 μM, respectively, and then cultured for 24 hours. Thereafter, the cultured cells were treated with the WST-1 reagent according to the manufacturer's instructions, and the absorbance at a wavelength of 450 nm was measured after a certain period of time. Here, the WST-1 reagent can determine the cell survival rate by changing the color of the culture medium according to the level of mitochondrial dehydrogenase present in the cells.

[0208] [4-2] Clonogenic assay

[0209] The breast cancer cell line of Experimental Method 1 was cultured at 1-2×10 3 The cells were distributed into a 6-well plate at a density of 10 cells / well. After the cells were fully attached, the cells were treated with the compound of Preparation Example 6 at a concentration of 0-30 μM and cultured for 10 to 14 days. Thereafter, the cultured cells were fixed with 4% formaldehyde and stained with 1% crystal violet, and clones were counted under a microscope.

[0210] [4-3] Flow cytometry

[0211] The breast cancer cell line of Experimental Method 1 was treated with the compound of Preparation Example 6 at a concentration of 0-30 μM and cultured for 24 hours. Afterwards, the cultured cells were detached using trypsin, and then double-stained with Annexin V (recognizing cell apoptosis) and PI (propidium iodide; identifying cell necrosis), and then the pattern and level of cell apoptosis were determined using flow cytometry.

[0212] [Experimental method 5] Methods for measuring epithelial-mesenchymal transition (EMT) in tumor cell lines

[0213] The culture medium of the breast cancer cell line in Experimental Method 1 was replaced with a serum-free medium. Subsequently, the cells were treated with 2 ng / mL of TGF-β1 and the compound of Preparation Example 6 and cultured for 20-24 hours. Thereafter, as described in Experimental Method 3, the protein expression level was determined using an antibody specific for EMT-related proteins.

[0214] [Experimental method 6] Construction of xenograft / orthotopic animal models of breast cancer cell lines

[0215] Purchase 4-week-old athymic nude mice (female), then make mice adapt to 7 days in the animal breeding room environment and only select healthy mice at the same time. To identify each mouse, use ear number clamps to mark the ear. In addition, the breeding box is identified by attaching a mouse identification card with test number, test substance name, test item, receipt date, test date, test content and test person in charge. The environment of the animal breeding room is maintained at a temperature of 22 ± 3 ° C, relative humidity of 50 ± 20%, 10-15 ventilation / hour, 12 hours of light (8: 30-20: 30) / dark cycle, light brightness 150-300lux. In addition, these animals are fed with feed and drinking water during feeding, and quarantined during adaptation and testing.

[0216] Nude mice were anesthetized by inhalation of 2.5% isoflurane and then injected with a sterile syringe into the mice with the breast cancer cell line 4T1-luc from Experimental Method 1. The cells were diluted in 100 μl PBS (phosphate buffered saline) so that 1-2.5×10 cells could be injected into each nude mouse. 6The cells were then injected into the flank or mammary fat pad. The composition of the xenograft animal model obtained by injecting the breast cancer cell line into the flank is shown in Table 2 below. In addition, the composition of the orthotopic animal model obtained by injecting the breast cancer cell line into the mammary fat pad is shown in Table 3 below.

[0217] [Table 2]

[0218]

[0219] [Table 3]

[0220]

[0221] At the same time, after eliminating abnormal mice, mice with abnormal weight gain, and unhealthy mice during the animal adaptation period, the mice were grouped to make the average weight and tumor size consistent.

[0222] [Experimental method 7] Method for evaluating the effect of the compound of Preparation Example 6 in an animal model

[0223] [7-1] Methods for evaluating efficacy in xenograft animal models

[0224] Tumor volume was measured daily from the day of injection of mouse breast cancer cell line 4T1-luc to the end of the experiment (dissection date). 3 The compound of Preparation Example 6 was injected into the tail vein of the animal model at a concentration of 4 mg / kg, once every 3 days, for a total of 6 times. In order to confirm the tumor regression effect, the volume of the tumor was measured before the administration of the compound of Preparation Example 6, and on the 28th day after the first administration of the compound of Preparation Example 6, the animals were euthanized, the primary tumor was excised and its volume was compared.

[0225] [7-2] Methods for evaluating the effect of increasing survival in in situ animal models

[0226] From the day of injection of mouse breast cancer cell line 4T1-luc to the end of the experiment (dissection date), tumor volume was measured every 3 days, and the survival rate of the animal model was checked every day. 3 The compound of Example 6 was injected into the abdominal cavity of the animal model at a concentration of 20 mg / kg, once every 3 days, for a total of 6 times. At the same time, for the dignity of the animal model, the experiment was carried out in the control group when the primary tumor volume reached 3000 mm 3 The experiment was terminated at the 32nd day and the animal model was euthanized.

[0227] [7-3] Methods for evaluating efficacy in in situ animal models

[0228] From the injection day of the mouse breast cancer cell line 4T1-luc cell line to the end of the experiment (dissection date), the lung metastasis of the tumor cell line was checked every 1-2 days. After the animal model was anesthetized with 2.5% isoflurane inhalation, the fluorescence intensity was measured using an in vivo imaging system (IVIS). The lung metastasis was checked in real time by measuring the position and number of the 4T1-luc cell line transferred to the lungs. From the first injection day of the compound of Preparation Example 6 to the end of the experiment (dissection date), 20 mg / kg of Preparation Example 6 compound was continuously injected intraperitoneally every 2 days.

[0229] result:

[0230] [Example 1] Evaluation of the effect of improving AMPK protein stability

[0231] The CTSA method described in Experimental Method 3 was used to detect whether the compound of Preparation Example 6 binds to the AMP binding site on AMPK. The results are as follows: Figure 1 shown.

[0232] like Figure 1 As shown, when AMPK protein was present alone, the protein level decreased from 50°C (T m When the cells were treated with the compound of Preparation Example 6, the AMPK protein level was maintained at up to 55°C (T m value is 55℃).

[0233] It can be seen from the above results that the compounds of the present invention can increase the thermal stability of AMPK protein by binding to the AMP binding site of AMPK.

[0234] [Example 2] Evaluation of the inhibitory effects on survival and growth of cancer cell lines and the ability to activate AMPK

[0235] [2-1] Evaluation of inhibitory effects on survival and growth of cancer cell lines

[0236] The effects of reducing the survival rate and inhibiting the growth of cancer cell lines were evaluated by cell viability assay (a) and monoclonal formation assay (b) described in Experimental Method 4. The results are shown in Figure 2a and 2b shown.

[0237] like Figure 2a and 2b As shown, when the cell line was treated with the positive control AICAR, the cell viability was inhibited by more than 50% at 1 mM, and when both 4T1 and BT549 cell lines were treated with 10 μM of the compound of Preparation Example 6, the cell viability was inhibited by more than 50% (see Figure 2aIn addition, when the cell lines were treated with 10 μM of the compound of Preparation Example 6, colony formation in 4T1, BT549 and MDA-MB-231 cell lines was significantly reduced (see Figure 2b ).

[0238] From the above results, it can be seen that the compounds of the present invention can reduce the survival rate of cancer cell lines and inhibit the growth of cancer cells.

[0239] [2-2] Evaluation of the effect of inhibiting the growth of cancer cell lines

[0240] The effect of inhibiting the growth of cancer cell lines was evaluated by the cell viability assay described in Experimental Method 4-1. Figure 3a and 4a -4c. Figure 3a As shown, it was verified that when various concentrations of each compound in a total of 14 compounds (including the compound of Preparation Example 6 and the structurally similar compounds (YE-01, 02, 03, 04, 05, 08, 09, 10, 11, 12, 13, 14 and 15) of Preparation Examples 1-5 and 8-15))) were used to treat breast cancer cell lines 4T1, BT549 and MDA-MB-231 for 24 hours, the compound of Preparation Example 6 or the compound of Preparation Example 8 was able to inhibit the survival rate of cancer cells by more than 50% in all three breast cancer cell lines at a concentration of 10 μM. In addition, it was verified that when the three cell lines of gastric cancer, brain cancer and pancreatic cancer were treated with various concentrations of the compound of Preparation Example 6 or the compound of Preparation Example 8 for 24 hours, the survival rate of cancer cells was effectively inhibited by more than 50% (see Figure 4a-4c ). From the above results, it can be seen that the compound of Preparation Example 6 of the present invention and the compound of Preparation Example 8 having a similar structure can more effectively reduce the survival rate of breast cancer cells and the survival rates of gastric cancer, brain cancer and pancreatic cancer cells.

[0241] [2-3] Evaluation of AMPK activation ability in cancer cell lines

[0242] According to the method described in the above experimental method 3, the AMPK activation ability in cancer cell lines was evaluated by the expression level of p-AMPK (Thr172). Figure 3b Here, the ratio of p-AMPK / AMPK was plotted by calculating the normalized value of each blot relative to the internal control (GAPDH). Figure 3bAs shown, it was verified that when the breast cancer cell lines 4T1, BT549 and MDA-MB-231 were treated with the compound of Preparation Example 6 and the compound of Preparation Example 8 for 24 hours, the compound of Preparation Example 6 and the compound of Preparation Example 8 effectively increased the p-AMPK level in the three cell lines. In addition, it was verified that when the three cell lines of gastric cancer, brain cancer and pancreatic cancer were treated with the compound of Preparation Example 6, the compound of Preparation Example 8 and AICAR at a concentration of 10 μM for 12 hours, compared with the untreated group (control group), the compound of Preparation Example 6 and the compound of Preparation Example 8 increased the expression of p-AMPK (see Figure 4a-4c ). Here, it can be seen that, compared with the compound of Preparation Example 6 and the compound of Preparation Example 8, the effect of AICAR used as a positive control is not significant, while compared with the AMPK activator AICAR, in addition to breast cancer cells, the compound of Preparation Example 6 of the present invention and the compound of Preparation Example 8 having a similar structure can more effectively induce the activation of AMPK in gastric cancer, brain cancer and pancreatic cancer cells.

[0243] [Example 3] Evaluation of the effect of inducing apoptosis in cancer cell lines

[0244] The effect of inducing apoptosis in cancer cell lines was detected by flow cytometry as described in Experimental Method 4 and Western blot as described in Experimental Method 3. Figure 5 and 6 shown.

[0245] like Figure 3b As shown, it was confirmed that when the 4T1 cell line and the MDA-MB-231 cell line were treated with the compound of Preparation Example 6, the apoptosis marker cleaved PARP (poly (ADP-ribose) polymerase) protein increased in a concentration-dependent manner along with the increase of phosphorylated AMPK protein, and the levels of Bcl-2 and Cyclin D1 proteins decreased.

[0246] like Figure 6 As shown, when the 4T1 cell line and the MDA-MB-231 cell line were treated with the compound of Preparation Example 6, the number of cells stained with Annexin V or PI increased in a concentration-dependent manner.

[0247] From the above results, it can be seen that the compounds of the present invention can induce apoptosis of cancer cells.

[0248] [Example 4] Evaluation of the effect of inhibiting the growth of cancer cell lines

[0249] The EMT of cancer cell lines was determined by the method described in Experimental Method 5, and the results were shown in Figure 7a and 7bHere, MMP2 (matrix metalloproteinase 2) and MMP9 (matrix metalloproteinase 9) are enzymes that degrade type IV collagen, which is an important component of the basement membrane and is directly related to cancer migration and metastasis. Therefore, the secretion levels of MMP2 and MMP9 proteins were detected.

[0250] like Figure 7a As shown, when BT549, MDA-MB-231 and 4T1 cell lines were treated with TGF-β1, the EMT protein markers α-SMA, Vimentin, ZEB-1 and Slug protein increased, and when these cell lines were treated with TGF-β1 in combination with the compound of Preparation Example 6, the EMT protein markers decreased.

[0251] like Figure 7b As shown, by treating with the compound of Preparation Example 6, the levels of secreted MMP2 and MMP9, which are EMT protein markers present in the BT549 cell line when treated with TGF-β1, were significantly reduced.

[0252] From the above results, it can be seen that the compounds of the present invention can very effectively inhibit cancer cell invasion and cancer metastasis by inhibiting the EMT of cancer cells.

[0253] [Example 5] Evaluation of tumor regression effects in animal models

[0254] The tumor regression effect of the compound of Preparation Example 6 was evaluated according to Method 7-1 of Experimental Method 7. The results are as follows Figure 8a-8c In addition, the effect of the compound of Preparation Example 6 on increasing the survival rate was evaluated according to Method 7-2 of Experimental Method 7, and the results were as follows Figure 8d shown.

[0255] like Figure 8a-8c As shown, in the animal model, the volume of the tumor in the group (YE-06) to which the compound of Preparation Example 6 was administered was reduced in a feeding time-dependent manner compared to the case where the compound of Preparation Example 6 was not administered (control) ( Figure 8a and 8b In addition, it was observed that the body weight did not decrease with feeding time in both the case where the compound of Preparation Example 6 was not administered (control) and the case where the compound of Preparation Example 6 was administered (YE-06) ( Figure 8c ).

[0256] In addition, if Figure 8d As shown, it was confirmed that the survival rate was significantly increased in the group (YE-06) to which the compound of Preparation Example 6 was administered, compared with the group (control) to which the compound of Preparation Example 6 was not administered.

[0257] From the above results, it can be seen that the compounds of the present invention are non-toxic in animal models, and the administration of the compounds can very effectively inhibit tumor growth. Further, it can be seen that in animal models, the compounds of the present invention can significantly increase the overall survival rate of cancer patients by effectively inhibiting tumor growth and metastasis.

[0258] [Example 6] Evaluation of the inhibitory effect on tumor metastasis in animal models

[0259] According to method 7-3 of experimental method 7, the tumor metastasis inhibition effect of the compound of preparation example 6 was evaluated. The results are as follows Figure 9a and 9b shown.

[0260] In addition, if Figure 9a and 9b As shown, it was verified that 4T1-luc metastasis was only observed in the lungs, liver, spleen and kidneys, and that very high fluorescence intensity was observed in the lungs of the in situ animal model when the compound of Preparation Example 6 was not administered (control), while in the case of administration of the compound of Preparation Example 6 (YE-06), the luminescence intensity in the lungs of the in situ animal model was significantly reduced.

[0261] It can be seen from the above results that in animal models, the compounds of the present invention can very effectively inhibit tumor metastasis, especially lung metastasis.

[0262] Although the present invention has been described in detail with reference to specific features, it is obvious to those skilled in the art that such description is only a preferred embodiment of the present invention and does not limit the scope of the present invention. Therefore, the essential scope of the present invention should be the scope defined by the attached claims and their equivalents.

[0263] Industrial Applicability

[0264] The composition of the present invention can not only be used to prevent, improve, or treat cancer very efficiently, but also can inhibit cancer metastasis by inhibiting the growth of cancer cells and very efficiently inhibiting the metastasis of cancer cells to other tissues.

Claims

1. Use of a pharmaceutical composition in the preparation of a therapeutic agent for preventing or treating cancer, the pharmaceutical composition comprising a compound represented by the following formula 1 as an active ingredient: [Formula 1] The cancer is at least one selected from the group consisting of gastric cancer, breast cancer, pancreatic cancer and brain cancer. in: L1 is phenylene; and L2 is C1 alkoxyphenylene; X is -NR'R"; and Y is C1 alkoxy; R' and R" are hydrogen and a substituent represented by the following formula 4: [Formula 4] R2 is halogen; and n and m are 1.

2. The use according to claim 1, which contains at least one compound selected from the following as an active ingredient:

3. The use according to claim 1, further comprising an anticancer drug.

4. Use of a pharmaceutical composition in the preparation of a therapeutic agent for inhibiting cancer metastasis, the pharmaceutical composition containing a compound represented by the following formula 1 as an active ingredient: [Formula 1] The cancer is at least one selected from the group consisting of gastric cancer, breast cancer, pancreatic cancer and brain cancer. in: L1 is phenylene; and L2 is C1 alkoxyphenylene; X is -NR'R"; and Y is C1 alkoxy; R' and R" are hydrogen and a substituent represented by the following formula 4: [Formula 4] R2 is halogen; and n and m are 1.

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