Novel cinnamamide derivatives and uses thereof

Abstract

The present invention relates to novel cinnamamide derivatives and their use in the prevention or treatment of cancer.

Description

[Technical Field]

[0001] The present invention relates to novel cinnamamide derivatives and their use in the prevention or treatment of cancer. [Background technology]

[0002] According to statistics released in May 2021 (Global Cancer Statistics 2020), there were 19.3 million new cancer cases worldwide in 2020, and the number of cancer-related deaths reached 10 million (H. Sung et al., CA Cancer J. Clin. 71,209,2021).

[0003] Since the first chemical anticancer drugs were used in the 1940s, various anticancer drugs have been developed with the goal of eradicating cancer cells and ultimately conquering cancer. First-generation anticancer drugs, developed until the mid-1990s, were drugs that suppressed the growth of rapidly growing cancer cells, but they had side effects that also affected normal cells. To address this issue, second-generation anticancer drugs, or targeted anticancer drugs, that selectively attack cancer cells were developed in the late 1990s. However, problems were discovered in which resistance developed over a certain period of time, resulting in a decrease in efficacy (P. Cohen et al. Nature Rev. Drug Discov. 20,551,2021).

[0004] To address this issue, third-generation anticancer drugs, or immune anticancer drugs, were developed in the 2010s. Immunotherapy is a treatment that inhibits or eliminates cancer growth by activating and suppressing the body's immune system. Examples of immunotherapy include immune checkpoint inhibitors, immune cell therapy, and therapeutic antibodies. Immune checkpoint inhibitors block the activation of immune checkpoint proteins involved in T cell suppression, activating T cells to attack cancer cells. Examples include CTLA-4, PD-1, and PD-L1 inhibitors. Immune cell therapy drugs include CAR T cell therapy, which strengthens and transforms T cells collected from the body using the autologous cell transplant (ACT) method, and then injects them to enhance cellular immunity against cancer cells. Natural killer cell (NK cell) therapy, which induces apoptosis of cancer cells by inducing a natural immune response, is also included. In addition, therapeutic antibodies include antibody-drug conjugates (ADCs), which are drugs that release drugs to attack cancer cells when the antibody-drug conjugate binds to the cancer cells (Y. Zhang, et al., Cell. Mol. Immunol. 17,807,2020).

[0005] Immunotherapy is a new anticancer drug that bridges the gap between chemotherapy and targeted therapy. It has achieved clinical success by dramatically increasing patient survival rates (AD Waldman, et al., Nature Rev. Immunol. 20, 651, 2020). However, the low immune reactivity of cancer cells is thought to be the cause of the low response rate to immune barrier inhibitors, among other immunotherapy drugs. In particular, the overall response rate is low in solid tumors (e.g., 15-20% in lung cancer), and side effects and resistance to immunotherapy have been reported, highlighting the limitations of immunotherapy (G. Morad, et al., Cell 184, 5309, 2021).

[0006] In a 2014 Nature article titled "Three known unknowns," Bourzac discussed three things about cancer that remain unclear despite progress in cancer treatment: anticancer drug resistance, cancer metastasis, and the role of normal cells surrounding cancer cells, i.e., the tumor microenvironment (K. Bourzac, Nature 509, S69, 2014). It is believed that only by finding answers to these three questions and developing regulators can humanity be freed from the disease of cancer.

[0007] Resistance to anticancer drugs can be broadly classified into two types: intrinsic resistance, in which factors that cause resistance to drugs are present in cancer cells from the beginning, causing the drug to not respond; and acquired resistance, in which mutations occur during the continuous administration of drugs after the drug has shown its anticancer effect, or the drug no longer responds due to interactions with other signaling systems.

[0008] There are numerous causes of resistance, but one area that has recently attracted attention is the tumor microenvironment (TME). Tumors themselves are highly heterogeneous and interact with a wide variety of cells in their surroundings. This is collectively referred to as the tumor microenvironment (TME). The tumor microenvironment contains not only tumor cells but also normal epithelial cells, endothelial cells, fibroblasts, and immune cells. These cells are organically connected to each other and are closely related to anticancer drug resistance (MR Junttila, Nature 501, 346, 2013). Immune cells that are important in the tumor microenvironment include regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), cancer-associated fibroblasts (CAFs), and natural killer cells (NK cells).

[0009] Meanwhile, signal transducer and activator of transcription 3 (STAT3) has been identified as an important protein in intracellular signal transduction, whose function is regulated by cytokines and hormones and is involved in cell growth and differentiation. Numerous studies have demonstrated that sustained STAT3 activity promotes tumor cell survival and metastasis (M. El-Tanani, et al., Cellular Signaling 92, 100275, 2022). Over the past decade, various studies have shown that STAT3 is involved in the induction of anticancer drug resistance and the maintenance of cancer stem cells (P. Shih, et al., Drug Discovery Today, 26, 1450, 2021). As previously mentioned, STAT3 is involved in regulating the function of immune cells, including regulatory T cells, myeloid-derived suppressor cells, and natural killer cells, in the tumor microenvironment, a major cause of anticancer drug resistance (D. Bayik, et al., Nature Rev. Cancer, 21, 526, 2021).

[0010] In this regard, there is an emerging need for research into the development of drugs that overcome anticancer drug resistance and enhance activity through anticancer immune activation, which converts the tumor microenvironment from cancers that do not respond to immune anticancer drugs (cold tumors) to cancers that do respond to immune anticancer drugs (hot tumors).

[0011] Under these circumstances, the present inventors have made intensive efforts to develop a method for more effectively treating or preventing cancer by regulating the growth of cancer cells and the cancer cell microenvironment. As a result, they synthesized a novel cinnamamide derivative and confirmed that the compound inhibits the growth of cancer cells and increases the activity of immune cells, which play an important role in the tumor microenvironment, by suppressing STAT3 activity, thereby completing the present invention. [Prior art documents] [Non-patent literature]

[0012] [Non-licensed document 1] H. Sung et al.,CA Cancer J. Clin. 71,209,2021 [Non-licensed document 2] P. Cohen et al. Nature Rev. Drug Discov. 20,551,2021 [Non-licensed document 3] Y. Zhang,et al.,Cell. Mol. Immunol. 17,807,2020

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

Non-licensed Document 13

[0013] The present inventors have developed novel cinnamamide derivatives that can be useful for the prevention or treatment of cancer, and have completed the present application. [Means for solving the problem]

[0014] One object of the present invention is to provide a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof:

[0015] [Chemical formula 1] JPEG2025525320000002.jpg3989

[0016] In the above Chemical Formula 1, R1 is C 1-6 Alkyl;Hydroxy;Halogen;C 1-6 alkynyloxy; or unsubstituted or substituted C 1-6 is alkoxy; The substituted C 1-6 Alkoxy is C 1-6 containing aryl or 5-14 membered heteroaryl as a substituent, the heteroaryl contains one or more O, N, or S; R2 is hydrogen or alkyl.

[0017] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0018] Another object of the present invention is to provide a method for preventing or treating cancer, which comprises administering to an individual other than a human a pharmaceutical composition containing, as an active ingredient, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0019] Another object of the present invention is to provide a food composition for preventing or ameliorating cancer, which comprises the compound represented by Chemical Formula 1 or a nutrient-acceptable salt thereof as an active ingredient.

[0020] Another object of the present invention is to provide a feed composition for preventing or improving cancer, which comprises the compound represented by Chemical Formula 1 or a nutrient-acceptable salt thereof as an active ingredient. [Effects of the Invention]

[0021] The novel compound of the present invention or a pharmaceutically acceptable salt thereof inhibits STAT3 activity, thereby exhibiting the effects of suppressing the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment, and is therefore useful for the prevention or treatment of cancer. [Brief explanation of the drawings]

[0022] [Figure 1] 1H-NMR data of compound CG165 according to an embodiment of the present invention. [Figure 2] FIG. 1 shows that Stat 3 phosphorylation is inhibited in a concentration-dependent manner in prostate cancer cell lines by CG165, a compound of the present invention. [Figure 3] FIG. 1 shows that CG165, a compound of the present invention, inhibits Stat 3 phosphorylation in a time-dependent manner in prostate cancer cell lines. [Figure 4] FIG. 1 shows the results of measuring the cell growth inhibitory activity of CG165, a compound of the present invention, in prostate cancer cell lines. [Figure 5] FIG. 1 shows the results of verifying that CG165, a compound of the present invention, has no cell growth inhibitory activity in normal cell lines. [Figure 6]FIG. 1 shows that CG165, a compound of the present invention, concentration-dependently inhibits Stat 3 phosphorylation in breast cancer and lung cancer cell lines. [Figure 7] FIG. 1 shows the results of measuring the cell growth inhibitory activity of CG165, a compound of the present invention, in breast cancer cell lines. [Figure 8] FIG. 1 shows that the compound of the present invention, CG165, suppresses the expression of Stat3 target proteins in cancer cell lines, thereby inducing cell death. [Figure 9] FIG. 1 shows that Stat 3 phosphorylation is suppressed by the compounds of the present invention, CG155 and CG165, in natural killer cells (NKL), which are immune cells. [Figure 10] FIG. 1 shows that Stat 3 phosphorylation is suppressed by the compounds of the present invention, CG155 and CG165, in bone marrow-derived macrophages (BMDMs), which are immune cells. [Figure 11] FIG. 1 shows the results of measuring the weight change of cancer tissue when CG165, a compound of the present invention, was orally administered to mice subcutaneously implanted with prostate cancer. [Figure 12] FIG. 1 shows the results of measuring the change in size of cancer tissue when CG165, a compound of the present invention, was orally administered to mice subcutaneously implanted with prostate cancer. [Figure 13] FIG. 1 shows the results of measuring changes in animal weight when CG165, a compound of the present invention, was orally administered to mice subcutaneously implanted with prostate cancer. [Figure 14] FIG. 1 shows the results of measuring the weight change of cancer tissue when CG126, a compound of the present invention, was intraperitoneally administered to mice subcutaneously implanted with prostate cancer. [Figure 15] FIG. 1 shows the results of a toxicity verification test of a single oral administration of CG165, a compound of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0023] This will be explained in more detail as follows. Meanwhile, each description and embodiment disclosed in the present invention also applies to each other description and embodiment. That is, any combination of various elements disclosed in the present application falls within the scope of the present invention. Furthermore, the following specific description is not considered to limit the category of the present application.

[0024] One aspect of the present invention provides a compound represented by the following formula 1, or a pharmaceutically acceptable salt thereof:

[0025] [Chemical formula 1] TIFF2025525320000003.tif3786

[0026] In the above Chemical Formula 1, R1 is C 1-6 Alkyl;Hydroxy;Halogen;C 1-6 alkynyloxy; or unsubstituted or substituted C 1-6 is alkoxy; The substituted C 1-6 Alkoxy is C 1-6 containing aryl or 5-14 membered heteroaryl as a substituent, the heteroaryl contains one or more O, N, or S; R2 is hydrogen or alkyl.

[0027] JPEG2025525320000004.jpg97170

[0028] For example, the compound of the present invention may be a compound having the following compound structure:

[0029] [Table 1] JPEG2025525320000006.jpg231148

[0030] Meanwhile, the compounds of the present invention can exist in the form of pharmaceutically acceptable salts. As salts, acid addition salts formed with pharmaceutically acceptable free acids are useful. The term "pharmaceutically acceptable salt" as used herein refers to any and all organic or inorganic addition salts of the compounds of Chemical Formula 1 that are relatively non-toxic to patients and whose side effects attributable to the salt do not diminish the beneficial effects of the compound at concentrations that have an innocuous effective effect.

[0031] Acid addition salts are prepared by conventional methods, for example, by dissolving the compound in an excess amount of aqueous acid and precipitating the salt with a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. Equal molar amounts of the compound and an acid or alcohol (e.g., glycol monomethyl ether) in water are heated, and the mixture is then evaporated to dryness, or the precipitated salt can be filtered off with suction.

[0032] In this regard, the free acid may be an organic acid or an inorganic acid. Examples of inorganic acids that may be used include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, and tartaric acid. Examples of organic acids that may be used include, but are not limited to, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid.

[0033] Pharmaceutically acceptable metal salts can also be prepared using bases. Alkali metal salts or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. In this case, sodium, potassium, or calcium salts are particularly suitable for pharmaceutical purposes, but are not limited to these. Corresponding silver salts can also be obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (e.g., silver nitrate).

[0034] Unless otherwise specified, pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compounds of Formula 1. For example, pharmaceutically acceptable salts may include sodium, calcium, and potassium salts of hydroxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate), and p-toluenesulfonate (tosylate) salts, which can be prepared by salt preparation methods known in the art.

[0035] The salt of the cinnamamide derivative compound of the present invention may be any cinnamamide derivative salt that is pharmaceutically acceptable and exhibits pharmacological activity equivalent to that of the cinnamamide derivative compound, without any limitation.

[0036] Furthermore, the compound represented by Chemical Formula 1 according to the present invention includes, without limitation, not only its pharmaceutically acceptable salts but also solvates such as hydrates that can be prepared therefrom and all possible stereoisomers. The solvates and stereoisomers of the compound represented by Chemical Formula 1 can be prepared from the compound represented by Chemical Formula 1 using methods known in the art.

[0037] Furthermore, the compound represented by Chemical Formula 1 according to the present invention may be prepared in a crystalline or amorphous form, and if prepared in a crystalline form, may optionally be hydrated or solvated. The present invention may include not only stoichiometric hydrates of the compound represented by Chemical Formula 1, but also compounds containing various amounts of water. The solvates of the compound represented by Chemical Formula 1 according to the present invention include both stoichiometric and non-stoichiometric solvates.

[0038] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0039] The compound represented by Chemical Formula 1, its pharmaceutically acceptable salts, and the prevention and treatment are as described above.

[0040] The term "active ingredient" as used herein means an ingredient that exhibits a desired activity alone, or that exhibits a desired activity together with a carrier or the like that is inactive by itself.

[0041] As used herein, the term "prevention" refers to any action that inhibits or delays the occurrence, spread, and recurrence of cancer-related diseases by administering the composition of the present invention, and "treatment" refers to any action that improves or beneficially alters the symptoms of the disease by administering the composition of the present invention.

[0042] Specifically, the pharmaceutical composition of the present invention can prevent or treat cancer-related diseases by inhibiting the activity of signal transducers and activators of transcription 3 (STAT3), but is not limited thereto.

[0043] The term "STAT 3" as used herein refers to a transcriptional regulator that regulates the expression of genes related to cell growth, differentiation, apoptosis, etc. by delivering signals corresponding to various cytokines and growth factors from outside the cell into cells. STAT 3 is known to contribute to the malignant progression of cancer cells through overexpression and non-ideal activation in various cancer cells (HQ Wang, et al., MedChem. 3, e124, 2022). Furthermore, activation of STAT 3 is known to weaken the anti-cancer immune effect by regulating the function of various immune cells, such as natural killer cells, in the tumor microenvironment (J. Huynh, Nature Rev. Cancer, 19, 82, 2019).

[0044] Specifically, the pharmaceutical composition of the present invention can prevent or treat cancer-related diseases by suppressing the activity of Stat 3 and regulating the activity of immune cells such as natural killer cells or bone marrow-derived macrophages, but is not limited thereto.

[0045] In one embodiment of the present invention, it was confirmed that the novel compounds of the present invention inhibit the phosphorylation of STAT3 in cancer cells, suppress the growth of cancer cells, inhibit the expression of STAT3 target genes and apoptosis genes, degrade apoptosis marker proteins, and induce apoptosis in cancer cells. They also inhibit the activity of STAT3 in natural killer cells and bone marrow-derived macrophages. It was confirmed that the novel compounds of the present invention inhibit the growth of cancer cells and increase the activity of immune cells in the tumor microenvironment by inhibiting STAT3 activity.

[0046] Specifically, the cancer that can be prevented or treated by the pharmaceutical composition of the present invention may be any one selected from the group consisting of colon cancer, pancreatic cancer, gastric cancer, liver cancer, breast cancer, cervical cancer, thyroid cancer, parathyroid cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, ovarian cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, colon cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulva carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, and pituitary adenoma, but is not limited thereto.

[0047] Specifically, the pharmaceutical composition according to the present invention may contain, as an active ingredient, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof in an amount of 0.0001 to 99.9 wt %, specifically 0.01 to 80 wt %, more specifically 1 to 10 wt %, based on the total weight of the final composition, but is not limited thereto.

[0048] The compositions of the present invention may further comprise a pharmaceutically acceptable carrier, diluent, or excipient, and may be formulated into various forms, such as oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, or sterile injection solutions, according to the intended purpose, and may be administered orally or via various routes, including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration. Examples of suitable carriers, excipients, or diluents contained in such compositions include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. The composition of the present invention may further contain fillers, anti-agglomerating agents, lubricants, wetting agents, flavorings, emulsifiers, preservatives, and the like.

[0049] Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are formulated by mixing the composition with at least one or more excipients, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

[0050] Oral liquid preparations include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to water and liquid paraffin, which are frequently used simple diluents.

[0051] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin. Injectables may contain conventional additives such as solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0052] The compositions of the present invention are administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment, without causing adverse effects. The effective dose level can be determined based on factors including the patient's health status, the type and severity of the disease, the activity of the drug, sensitivity to the drug, the method, time, route and excretion rate of administration, duration of treatment, coadministered or concurrently used drugs, and other factors well known in the medical field. The compositions of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, in single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that will achieve maximum efficacy at the minimum dose without adverse effects, which can be easily determined by one of ordinary skill in the art.

[0053] For example, the dosage may be increased or decreased depending on the route of administration, severity of the disease, sex, body weight, age, etc., and therefore the above dosage does not limit the scope of the present invention in any way.

[0054] Specifically, the dosage of the pharmaceutical composition of the present invention may be, for example, 1 to 10 mg / kg body weight per day to animals, including humans, but is not limited thereto. The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be once a day or divided into several doses and administered several times a day. However, since the dosage may be increased or decreased depending on the route of administration, severity of the disease, sex, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0055] Specifically, the composition may further comprise an anti-cancer agent, but is not limited thereto.

[0056] More specifically, the anti-cancer agent may be at least one selected from the group consisting of DNA alkylating agents, anti-cancer antibiotics, plant alkaloids, targeted anti-cancer agents, and cancer immunotherapy, but is not limited thereto.

[0057] More specifically, the anticancer drug may be mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (actinomycin D), doxorubicin (doxorubicin: adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin, C bleomycin (C Bleomycin; vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, irinotecan, Glivec, Tasigna, Herceptin, Iressa, Nexavar, Yervoy, Opdivo, Keytruda, Tecentriq, Imfinzi, and Bavencio, but are not limited thereto.

[0058] Among the many causes of anticancer drug resistance, the tumor microenvironment has recently attracted considerable attention. Tumors themselves are highly heterogeneous and interact with a wide variety of cells in their surroundings. This is collectively referred to as the tumor microenvironment (TME). The tumor microenvironment includes not only tumor cells but also normal epithelial cells, endothelial cells, fibroblasts, and immune cells. These cells are interconnected and closely related to anticancer drug resistance (MR Junttila, Nature 501, 346, 2013). Immune cells important in the tumor microenvironment include regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), cancer-associated fibroblasts (CAFs), and natural killer cells (NK cells).

[0059] In this regard, activation of Stat 3 is known to regulate the function of various immune cells, including natural killer cells, in the tumor microenvironment, thereby reducing the immune anti-cancer effect (H. Yu, Nature Reviews Immunology 7, 41, 2007).

[0060] The novel compound of the present invention or a pharmaceutically acceptable salt thereof exhibits the effects of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by suppressing STAT3 activity, and can therefore be usefully used as a drug that overcomes resistance to anticancer drugs and enhances their activity through anticancer immune activation, which converts the tumor microenvironment from a cancer that does not respond to immune anticancer drugs (cold tumor) to a cancer that responds to immune anticancer drugs (hot tumor).

[0061] Another aspect of the present invention provides a method for preventing or treating cancer, comprising administering to an individual a pharmaceutical composition comprising, as an active ingredient, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0062] The compound represented by Chemical Formula 1, its pharmaceutically acceptable salt, active ingredient, pharmaceutical composition, cancer prevention and treatment are as described above.

[0063] As used herein, the term "individual" may refer to any animal, including, but not limited to, cattle, horses, sheep, pigs, goats, camels, antelopes, dogs, cats, and humans, which has or may develop cancer.

[0064] The prevention or treatment method of the present invention may specifically include the step of administering a pharmaceutically effective amount of the composition to an individual who has or is at risk of developing cancer.

[0065] The term "administration" as used herein means introducing the composition of the present invention into an individual who exhibits or is at risk of exhibiting symptoms of cancer and related diseases by any appropriate method. The composition of the present invention can be administered via various routes, such as oral or parenteral, as long as it can be delivered to the target tissue.

[0066] Specifically, the composition of the present invention can be administered via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, intranasal administration, intrapulmonary administration, and intrarectal administration, depending on the purpose, although this is not particularly limited thereto.

[0067] Specifically, the dosage of the pharmaceutical composition of the present invention may be, for example, 1 to 10 mg / kg body weight per day to animals, including humans, but is not limited thereto. The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be once a day or divided into several doses and administered several times a day. However, since the dosage may vary depending on the route of administration, severity of the disease, sex, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0068] Specifically, the pharmaceutical composition can prevent or treat cancer-related diseases by inhibiting the activity of signal transducers and activators of transcription 3 (STAT3), but is not limited thereto.

[0069] Another aspect of the present invention provides a food composition for preventing or ameliorating cancer, which comprises the compound represented by Chemical Formula 1 or a nutrient-acceptable salt thereof as an active ingredient.

[0070] The compound represented by Chemical Formula 1, the active ingredient, cancer and prevention are as described above.

[0071] The term "food-based acceptable salt" as used herein is as defined above in the term "pharmaceutically acceptable salt".

[0072] The term "amelioration" in the present invention means any action that at least reduces the parameters associated with the condition treated by administration of a composition according to the present invention, for example the severity of the symptoms.

[0073] The term "food" as used herein includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, drinking water, tea, energy drinks, alcoholic beverages, vitamin complexes, functional health foods and health foods, and includes all foods in the ordinary sense.

[0074] The food composition of the present invention can be taken on a daily basis and is expected to have a significant effect in improving cancer, making it extremely useful for the purpose of promoting health.

[0075] The food composition of the present invention can be used as a functional health food. The term "functional health food" refers to a food manufactured and processed using ingredients or components that have beneficial functions for the human body, as defined by the Health Functional Food Act. "Functional" refers to the food being consumed for the purpose of obtaining beneficial health effects, such as regulating nutrients for the structure and function of the human body or achieving physiological effects. The food of the present invention can be manufactured using methods commonly used in the art, and can be prepared by adding ingredients and components commonly used in the art. The food can be prepared in any dosage form that is recognized as a food. The food composition of the present invention can be manufactured into various dosage forms, and unlike common pharmaceuticals, it is made from natural ingredients, eliminating side effects that can occur with long-term drug administration. Furthermore, because of its portability, the food of the present invention can be taken as an adjuvant to enhance the effects of improving Parkinson's disease.

[0076] The term "health food" refers to food that has more active health maintenance and promotion effects than general foods, and "health supplement food" refers to food intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement" are used interchangeably.

[0077] Specifically, the functional health food is a food product obtained by adding the extract of the present invention to food ingredients such as beverages, teas, spices, gum, and confectioneries, or by encapsulating, powdering, or preparing a suspension, etc., and when ingested, it brings about specific health benefits. However, unlike general medicines, it has the advantage of being made from food ingredients and therefore free from side effects that can occur when taking medicines for a long period of time.

[0078] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

[0079] The food composition may also contain additional ingredients commonly used in food compositions to enhance the sense of smell, taste, or vision, such as vitamins A, C, D, E, B1, B2, B6, and B12, niacin, biotin, folate, and panthotenic acid. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr) may also be included, as well as amino acids such as lysine, tryptophan, cysteine, and valine.

[0080] The food compositions may also contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high-strength bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclomate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium D-bitartrate, etc.), fortifiers, emulsifiers, thickeners (thickeners), coating agents, gum bases, foam inhibitors, solvents, and improvers. The additives may be selected depending on the type of food and used in appropriate amounts.

[0081] The food composition of the present invention may contain common food additives, and unless otherwise specified, the suitability of the food composition as a "food additive" is determined in accordance with the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety.

[0082] Examples of the items listed in the "Food Additives Code" include chemically synthesized substances such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon color, licorice extract, crystalline cellulose, sorghum color, and guar gum; and mixed preparations such as monosodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0083] The food composition of the present invention may contain 0.01 to 95 wt %, preferably 1 to 80 wt %, of the compound of Chemical Formula 1 or a nutritively acceptable salt thereof based on the total weight of the composition. In addition, the compound of Chemical Formula 1 or a nutritively acceptable salt thereof contained in the food composition of the present invention may be obtained by the same or similar extraction method as mentioned in the preparation of the pharmaceutical composition, but is not limited thereto.

[0084] Furthermore, the food composition of the present invention can be manufactured and processed into the form of tablets, capsules, powder, granules, liquid, pills, etc. for the purpose of preventing or ameliorating cancer.

[0085] For example, the tablet-shaped health functional food may be prepared by granulating a mixture of the compound of Chemical Formula 1 or its nutrient-acceptable salt, excipient, binder, disintegrant, and other additives in a conventional manner, and then compressing the mixture with a lubricant, etc. Alternatively, the tablet-shaped health functional food may be directly compressed and molded. The tablet-shaped health functional food may also contain a flavoring agent, etc., if necessary, and may be coated with a suitable coating agent, if necessary.

[0086] Among capsule-type health functional foods, hard capsules can be produced by filling a powder mixture of the compound of Chemical Formula 1 or its nutrient-acceptable salt and additives such as excipients, or granules or coated granules thereof, into a conventional hard capsule, while soft capsules can be produced by filling a mixture of the compound of Chemical Formula 1 or its nutrient-acceptable salt and additives such as excipients into a capsule base such as gelatin. The soft capsules can contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., as needed.

[0087] The round health functional food can be prepared by molding a mixture of the compound of Chemical Formula 1 or its nutrient-acceptable salt, excipients, binders, disintegrants, etc. in an appropriate manner, and if necessary, the compound can be coated with white sugar or other appropriate coating agents, or coated with starch, talc, or other appropriate substances.

[0088] The granular health functional food can be prepared by a suitable method using a mixture of the compound of Chemical Formula 1 or its nutrient-acceptable salt, excipients, binders, disintegrants, etc., and can contain flavoring agents, flavoring agents, etc., as needed. When the granular health functional food is subjected to the following particle size test using sieves No. 12 (1680 μm), No. 14 (1410 μm), and No. 45 (350 μm), the amount that passes through the No. 12 sieve and remains on the No. 14 sieve may be 5.0% or less of the total amount, and the amount that passes through the No. 45 sieve may be 15.0% or less of the total amount.

[0089] The definitions of the terms excipient, binder, disintegrant, lubricant, flavoring agent, flavoring agent, etc. are those described in documents known in the art, and may include substances whose functions are the same or similar (Commentary on the Korean Pharmacopoeia, Moonsung Publishing, Korea Pharmaceutical University Association, 5th Revised Edition, pp. 33-48, 1989).

[0090] Another aspect of the present invention provides a feed composition for preventing or ameliorating cancer, which comprises the compound represented by Chemical Formula 1 or a nutrient-acceptable salt thereof as an active ingredient.

[0091] The compound represented by Chemical Formula 1, its nutrient-acceptable salt, active ingredient, and cancer prevention and amelioration are as described above.

[0092] The term "feed" as used herein means food ingested by animals, specifically, a substance that provides organic or inorganic nutrients necessary to sustain the life of an animal or to produce meat, milk, etc. The feed may contain feed additives and may be prepared in various forms known in the art.

[0093] The composition comprising the compound of Formula 1 or a nutrient-acceptable salt thereof of the present invention can be used to prevent or treat cancer in non-human individuals such as livestock or pets, and can be used as a functional feed additive or a feed composition.

[0094] The content of the compound of Chemical Formula 1 or its nutrient-acceptable salt in the feed composition according to the present invention can be appropriately adjusted depending on the type and age of the livestock to which it is applied, the application method, the desired effect, etc., and can be, for example, 1 to 99 wt %, preferably 10 to 90 wt %, more preferably 20 to 80 wt %, but is not limited thereto.

[0095] The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, oils and fats, starches, lees, or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0096] Alternatively, the compound of the present invention or a nutrient-acceptable salt thereof can be used as a feed additive to be added to a feed composition. The feed additive may be, but is not limited to, for improving the productivity or health of the target animal. The feed additive may be a supplemental feed under the Feed Control Act.

[0097] The feed additive of the present application can further contain one or more of organic acids such as citric acid, fumaric acid, adipic acid, and lactic acid, and natural antioxidants such as polyphenols, catechin, tocopherol, vitamin C, green tea extract, chitosan, and tannic acid, and can also contain other common additives such as buffers and bacteriostatic agents as needed. Furthermore, the feed additive can be formulated into liquid, capsule, granule, or tablet form as needed.

[0098] The feed or feed additive may further contain substances that have various effects, such as supplementing nutrients and preventing weight loss, increasing the digestibility of fiber in the feed, improving milk quality, preventing reproductive disorders and increasing conception rates, preventing heat stress in summer, etc. For example, in addition to main ingredients such as various supplements such as amino acids, inorganic salts, vitamins, antioxidants, antifungals, and microbial preparations, and plant protein feeds such as crushed or shredded wheat, barley, and corn, animal protein feeds such as blood meal, meat meal, and fish meal, and animal and plant fats, the feed or feed additive may also contain nutrient supplements, growth promoters, digestion and absorption promoters, and disease preventive agents.

[0099] The feed and feed additives of the present application can be fed to a wide variety of animals, including mammals, poultry, and, for example, but not limited to, livestock such as cows, goats, and pets such as dogs and cats to maintain or improve their appearance.

[0100] Another aspect of the present invention provides a use of a composition comprising the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient for preventing or treating cancer.

[0101] The compound represented by Chemical Formula 1, its pharmaceutically acceptable salt, active ingredient, cancer, prevention and treatment are as described above.

[0102] The present invention will be described in more detail below with reference to examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0103] Synthesis Example 1: Synthesis of CG133 Among the compounds of Chemical Formula 1, CG133 was synthesized according to the following reaction scheme 1.

[0104] [Reaction Scheme 1] JPEG2025525320000007.jpg107137

[0105] Specifically, methyl 2-hydroxycinnamate (5 g) was dissolved in methylene chloride (200 mL) and then 3,4-dihydropyran (2.6 g, 1.1 equivalents) was added. A trace amount of p-toluenesulfonic acid (5-10 mg) was added to the reaction solution and stirred at room temperature for 4 hours. A small amount of triethylamine (0.5 mL) was added to terminate the reaction. The solvent was then concentrated under reduced pressure and extracted three times with ethyl acetate (EA) and water. The organic layer was then dried over MgSO4, filtered, and concentrated under reduced pressure. The concentrate was used in the next reaction without further purification.

[0106] The concentrate and methacrylamide (2.7 g, 1.1 equivalents) were dissolved in DMF (N,N-dimethylformamide, 100 mL). Sodium hydride (800 mg, 1.2 equivalents) was added to the mixture and stirred at room temperature for 7 hours. 50 mL of saturated ammonium chloride solution and water were added to terminate the reaction, followed by extraction three times with ethyl acetate and water. The organic layer was then dried over MgSO4, filtered, and concentrated under reduced pressure. The concentrate was dissolved in 100 mL of methyl alcohol, and p-toluenesulfonic acid (20 mg) was added. The mixture was stirred at room temperature for 4 hours. The solvent was then concentrated under reduced pressure, followed by extraction three times with ethyl acetate (EA) and water. The organic layer was then dried over MgSO4, filtered, and concentrated under reduced pressure. The concentrate was purified using column chromatography (EA:Hexane = 20:80) to obtain compound CG133 (3.1 g, 47.7%, light yellow powder).

[0107] The mp of compound CG133 is 198°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0108] 1 HNMR (400 MHz, CDCl3)δ9.38 (brs, 1H), 9.30 (brs, 1H), 7.85 (d, J = 16 Hz, 1H), 7.29 (d, J = 16 Hz, 1H), 7.25 (d, J = 7.6 Hz, 1H), 6.93 (t, J = 7.6 Hz, 1H), 6.65 (d, J = 7.6 Hz, 1H), 6.57 (t, J = 7.6 Hz, 1H), 5.67 (s, 1H), 5.35 (s, 1H), 1.74 (s, 3H); 13 CNMR (100 MHz, CDCl3)δ167.08, 166.96, 156.55, 140.48, 139.26, 130.86, 128.32, 121.82, 121.21, 119.02, 118.85, 115.87, 17.93.

[0109] Synthesis Example 2: Synthesis of CG164 Among the compounds of Chemical Formula 1, CG164 was synthesized in the same manner as in Reaction Scheme 2 below.

[0110] [Reaction Scheme 2] JPEG2025525320000008.jpg104122

[0111] Specifically, methyl 2-hydroxycinnamate (5 g) was dissolved in acetone (200 mL), and potassium carbonate (4.7 g, 1.2 equivalents) was added to the solution. Propargyl bromide (4.0 g, 1.2 equivalents) was added to the reaction solution, and the solution was stirred at 50-60°C for 6 hours. Upon completion of the reaction, the reaction solution was stirred at room temperature for 1 hour, filtered, and concentrated under reduced pressure. The concentrated solution was used in the next reaction without further purification.

[0112] The concentrate and methacrylamide (2.7 g, 1.1 equivalents) were dissolved in DMF (N,N-dimethylformamide, 100 mL). Sodium hydride (800 mg, 1.2 equivalents) was then added to the mixture and stirred at room temperature for 7 hours. The reaction was terminated by adding 50 mL of saturated ammonium chloride solution and water, followed by extraction three times with ethyl acetate and water. The organic layer was then dried over MgSO4, filtered, and concentrated under reduced pressure. The concentrate was purified using column chromatography (EA:Hexane = 15:85) to obtain compound CG164 (3.8 g, 50.4%, white powder).

[0113] The mp of compound CG164 is 122°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0114] 1HNMR (600 MHz, CDCl3)δ8.32 (brs, 1H), 8.25 (d, J = 16 Hz, 1H), 7.76 (d, J = 16 Hz, 1H), 7.71 (dd, J = 1.8, 7.8 Hz, 1H), 7.40 (t, J = 7.8 Hz, 1H), 7.08 (d, J = 7.6 Hz, 1H), 7.04 (t, J = 7.8 Hz, 1H), 5.89(s, 1H), 5.65 (s, 1H), 4.81 (d, J = 2.4 Hz, 2H), 2.53 (t, J = 2.4 Hz, 1H), 2.05 (s, 3H); 13 CNMR (150 MHz, CDCl3)δ167.61, 166.60, 156.45, 141.31, 139.93, 131.74, 128.92, 124.26, 122.50, 121.68, 119.73, 112.79, 78.19, 76.01, 56.17, 18.46.

[0115] Synthesis Example 3: Synthesis of CG165 Among the compounds of Chemical Formula 1, CG165 was synthesized by the following method.

[0116] Specifically, a compound was synthesized by reacting CG164 synthesized in Synthesis Example 2 with methyl iodide in acetone in the presence of potassium carbonate (K2CO3), and named CG165. 1 The H-NMR data is shown in Figure 1.

[0117] The mp of compound CG165 is 115°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0118] 1HNMR (600 MHz, CDCl3)δ7.87(d, J = 15.6 Hz, 1H), 7.47 (dd, J = 1.8, 7.8 Hz, 1H), 7.35 (dt, J = 1.8, 7.8 Hz, 1H), 7.04 (d, J = 7.8 Hz, 1H), 7.01 (t, J = 7.8 Hz, 1H), 6.86 (d, J = 15.6 Hz, 1H), 5.43 (m, 2H), 4.77 (d, J = 2.4 Hz, 2H), 3.30 (s, 3H), 2.53 (t, J = 2.4 Hz, 1H), 2.06 (d, J = 1.2 Hz, 3H); 13 CNMR (150 MHz, CDCl3)δ174.85, 170.07, 156.40, 142.26, 138.41, 131.30, 129.52, 124.15, 122.34, 121.82, 121.71, 112.73, 78.06, 76.07, 56.09, 32.62, 19.05.

[0119] Synthesis Example 4: Synthesis of CG126 Among the compounds of Chemical Formula 1, CG126 was synthesized by the following method.

[0120] Specifically, CG126 was synthesized using 2-methoxycinnamic acid as a starting material in the same manner as in Reaction Scheme 2 above.

[0121] The mp of compound CG126 is 126°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0122] 1HNMR (500 MHz, CDCl3) δ8.22(d, J = 15.5 Hz, 1H), 8.20 (brs, 1H), 7.75 (d, J = 15.5 Hz, 1H), 7.64 (dd, J = 1.5, 7.5 Hz, 1H), 7.36 (dt, J = 1.5, 7.7 Hz, 1H), 6.96 (t, J = 7.5 Hz, 1H), 6.92 (d, J = 8.0 Hz, 1H), 5.85 (s, 1H), 5.62 (m, 1H), 3.90 (s, 3H), 2.03 (s, 3H); 13 CNMR (100 MHz, CDCl3) δ167.61, 166.47, 158.66, 141.87, 140.00, 131.96, 129.12, 123.60, 122.34, 120.68, 119.32, 111.14, 55.53, 18.45.

[0123] Synthesis Example 5: Synthesis of CG127 Among the compounds of Chemical Formula 1, CG127 was synthesized by the following method.

[0124] Specifically, CG127 was synthesized using 2-methylcinnamic acid as a starting material in the same manner as in Reaction Scheme 2.

[0125] The mp of compound CG127 is 135°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0126] 1HNMR (500 MHz, CDCl3) δ8.21 (brs, 1H), 8.18(d, J = 16 Hz, 1H), 7.69 (d, J = 7.5 Hz, 1H), 7.66 (d, J = 15.5 Hz, 1H), 7.29 (dt, J = 1.5, 7.5 Hz, 1H), 7.23 (m, 2H), 5.87 (s, 1H), 5.64 (m, 1H), 2.47 (s, 3H), 2.04 (s, 3H); 13 CNMR (100 MHz, CDCl3) δ167.32, 166.46, 144.16, 139.87, 138.18, 133.47, 130.81, 130.44, 127.00, 126.35, 122.57, 120.08, 18.85, 18.42.

[0127] Synthesis Example 6: Synthesis of CG155 Among the compounds of Chemical Formula 1, CG155 was synthesized by the following method.

[0128] Specifically, CG133 produced in Synthesis Example 1 above was reacted with 4-bromomethylpyridine in the presence of sodium hydride to synthesize CG155.

[0129] The mp of compound CG155 is 156°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0130] 1HNMR (600 MHz, CDCl3) δ8.65 (s, 2H), 8.44 (brs, 1H), 8.30 (d, J = 15.6 Hz, 1H), 7.82 (d, J = 15.6 Hz, 1H), 7.71 (dd, J = 1.2, 7.8 Hz, 1H), 7.41 (d, J = 4.8 Hz, 2H), 7.34 (dt, J = 1.2, 8.4 Hz, 1H), 7.03 (t, J = 7.8 Hz, 1H), 6.89 (d, J = 7.8 Hz, 1H), 5.90 (s, 1H), 5.65 (d, J = 1.2 Hz, 1H), 5.21 (s, 2H), 2.05 (s, 3H); 13 CNMR (150 MHz, CDCl3) δ176.66, 166.55, 156.97, 150.02, 145.74, 141.18, 139.88, 131.89, 129.42, 124.13, 122.59, 121.61, 121.56, 120.11, 112.46, 68.63, 18.45.

[0131] Synthesis Example 7: Synthesis of CG168 Among the compounds of Chemical Formula 1, CG168 was synthesized by the following method.

[0132] Specifically, CG168 was synthesized using 2-chlorocinnamic acid as a starting material in the same manner as in Reaction Scheme 2.

[0133] The mp of compound CG168 is 147°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0134] 1HNMR (600 MHz, CDCl3) δ8.55 (brs, 1H), 8.29 (d, J = 16.2 Hz, 1H), 7.79 (dd, J = 1.2, 7.8 Hz, 1H), 7.76 (d, J = 15.6 Hz, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.31 (m, 2H), 5.94 (s, 1H), 5.69 (d, J = 1.2 Hz, 1H), 2.06 (s, 3H); 13 CNMR (150 MHz, CDCl3) δ167.12, 166.71, 141.98, 139.68, 135.40, 132.84, 131.35, 130.16, 128.11, 127.08, 122.94, 121.79, 18.45.

[0135] Synthesis Example 8: Synthesis of CG187 Among the compounds of Chemical Formula 1, CG187 was synthesized by the following method.

[0136] Specifically, CG187 was synthesized using 2-hydroxycinnamic acid as a starting material in the same manner as in Reaction Scheme 2.

[0137] The mp of compound CG187 is 127°C, 1 H, 13 The results of the C-NMR analysis are as follows:

[0138] 1 HNMR (600 MHz, CDCl3) δ8.43 (brs, 1H), 8.31 (d, J = 15.6 Hz, 1H), 7.76 (d, J = 16.2 Hz, 1H), 7.70 (dd, J = 1.2, 7.8 Hz, 1H), 7.5-7.3 (m, 6H), 6.98 (m, 2H), 5.89 (s, 1H), 5.62 (d, J = 1.2 Hz, 1H), 5.20 (s, 2H), 2.04 (s, 3H); 13CNMR (100 MHz, CDCl3) δ167.70, 166.55, 157.65, 141.56, 139.90, 136.59, 131.89, 120.01, 128.69, 128.64, 127.99, 127.77, 127.36, 124.06 122.47, 121.07, 121.04, 119.63, 112.73, 70.42, 18.45.

[0139] Experimental Example 1: Cell Culture All cell lines used to verify the in vitro activity of the compounds prepared in Synthesis Examples 1 to 8 were purchased from ATCC (Manassas, VA, USA). Prostate cancer cell line DU-145, breast cancer cell line MDA-MB-468, lung cancer cell line HCC827, and human foreskin fibroblasts (HFF) were incubated in RPMI-1640 (Gibco, Gaithersburg, MD, USA) medium supplemented with 10% FBS (Gibco) and 100 μg / ml penicillin. Human breast epithelial cells (MCF10A) were incubated in DMEM-F12 (Gibco) medium supplemented with 10% fetal bovine serum (FBS), 0.5% penicilin / streptomycin, EGF (20 ng / ml), insulin (10 μg / ml), and hydrocortisone (0.5 mg / ml). The natural killer cell (NKL) cell line was grown in RPMI-1640 (ATCC, 30-2001) medium supplemented with 10% FBS (Gibco) and 50 ng / ml hIL-2 (Peprotech, East Windsor, NJ, USA). All cell lines used in the experiments were maintained in a 5% CO incubator.

[0140] Experimental Example 2: Western blot analysis Prostate cancer cell line DU-145, breast cancer cell line MDA-MB 468, lung cancer cell line HCC827, immune cell natural killer cells (NKL), or bone marrow-derived macrophages (BMDM) (4 × 10 5Cells) were plated in a 6-well plate. After 24 hours, the cells were treated with 0.1% DMSO, HCA, EA, and the compounds prepared in Synthesis Examples 1 to 8 at various concentrations and for various time periods. After washing with PBS, the cells were harvested using RIPA buffer. 20 μg of protein quantified using a Bio-Rad protein assay kit (Bio-Rad, Hercules, California, USA) was subjected to 8% SDS-PAGE electrophoresis and transferred onto a PVDF membrane (EMD Millipore, Billerica, Massachusetts, USA). The membrane was blocked with 5% skim milk in TBST, and experiments were performed using primary and secondary antibodies. The primary antibodies used were p-STAT3 (Y705) (Cell Signaling), STAT3 (Cell Signaling), Cyclin D1 (Santa Cruz), Cyclin A (Santa Cruz), Mcl-1 (Santa Cruz), Survivin (Cell Signaling), Bcl-xL (Santa Cruz), PARP (Cell Signaling), and GAPDH (Santa Cruz). The secondary antibodies used were HRP-conjugated goat anti-rabbit and anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). After washing three times with TBST, bands were detected using Luminata Forte Western HRP substrate (EMD Millipore) reagent and an LAS 4000 mini (GE Healthcare Life Sciences) instrument. Subsequently, correction using GAPDH was carried out using the MultiGauge program (Fuji Photo Film, Tokyo, Japan).

[0141] Example 1: Confirmation of the inhibitory effect of Stat 3 phosphorylation in cancer cell lines - 1 The inhibitory effect of the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8 on Stat 3 phosphorylation in cancer cell lines was confirmed by Western blotting.

[0142] Specifically, we decided to confirm the inhibitory effect on Stat 3 phosphorylation in a colon cancer cell line, which is a representative example of cancer.

[0143] The results are shown in Table 2 below.

[0144] [Table 2] JPEG2025525320000010.jpg185141 JPEG2025525320000011.jpg83138

[0145] As a result, it was confirmed that the cinnamamide derivative compounds of Formula 1 prepared in Synthesis Examples 1 to 8 of the present invention could inhibit STAT3 activity (p-STAT3) by 50% or more.

[0146] Next, the inhibitory effect of the compound of the present invention, CG165, on Stat 3 phosphorylation in cancer cell lines was confirmed by Western blotting.

[0147] As a result, as shown in Figures 2 and 3, it was confirmed that treatment with the compound CG165 of the present invention inhibited Stat 3 activity (p-STAT3) in the prostate cancer cell line DU-145 in a concentration-dependent manner, and in particular, it was confirmed that treatment with 2 μM inhibited Stat 3 activity (p-STAT3) by more than 50% (Figure 2).

[0148] Furthermore, it was confirmed that treatment with the compound of the present invention, CG165, inhibited Stat 3 activity (p-STAT3) in a time-dependent manner in the prostate cancer cell line DU-145 (FIG. 3).

[0149] Example 2: Confirmation of the cancer cell growth inhibitory effect of suppressing Stat 3 phosphorylation in cancer cell lines - 1 In Example 1, it was confirmed that Stat 3 activity (p-STAT3) was inhibited in cancer cells by treatment with the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8. To confirm the effect of inhibiting Stat 3 activity on the inhibition of cancer cell growth, a cell proliferation assay was performed using a prostate cancer cell line, a representative carcinoma.

[0150] Specifically, the prostate cancer cell line DU-145 (0.8x10 4 Cells were plated in a 96-well plate. After 24 hours, the cells were treated with DMSO (control) or CG-165 at various concentrations for 24 or 48 hours, followed by treatment with 10 μl of WST-1 (Dojindo Laboratories, Kumamoto, Japan) in each well. After approximately 1 hour of treatment, the absorbance was measured at 450 nm using a microplate reader (Multiskan GO system, Thermo Fisher Scientific, Rockford, IL, USA) and cell viability was calculated using the values.

[0151] As a result, as shown in FIG. 4, it was confirmed that the compound of the present invention, CG165, inhibits the growth of cancer cells in a concentration-dependent manner.

[0152] Therefore, it was confirmed that the compound of the present invention, CG165, inhibits the activity of Stat 3 in cancer cells, thereby inhibiting the growth of cancer cells.

[0153] Example 3: Confirmation of the effect of the compound of the present invention on normal cells To confirm that the compound CG165 of the present invention does not affect the growth of normal cells, human foreskin fibroblasts (HFF) and human breast epithelial cells (MCF10A) were treated with CG-165 at different concentrations for 48 hours, and then cell viability was measured using the WST-1 (Dojindo Laboratories, Kumamoto, Japan) method.

[0154] As a result, as shown in FIG. 5, it was confirmed that the compound of the present invention, CG165, did not significantly affect the growth of normal cells even when treated at a concentration of 10 μM for 48 hours.

[0155] Therefore, it was confirmed that the compound CG165 of the present invention selectively inhibits the growth of cancer cells while having little effect on normal cells.

[0156] Example 4: Confirmation of the inhibitory effect of Stat 3 phosphorylation in cancer cell lines - 2 The inhibitory effect of the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8 on Stat 3 phosphorylation was also confirmed by Western blotting in breast cancer cell lines and lung cancer cell lines, which are representative examples of cancers other than prostate cancer cell lines.

[0157] Specifically, the inhibitory effect of CG165 of the present invention on Stat 3 phosphorylation was confirmed by Western blotting in breast cancer cell line MDA-MB-468 and lung cancer cell line HCC827.

[0158] As a result, as shown in Figure 6, when the breast cancer cell line MDA-MB-468 and the lung cancer cell line HCC827 were treated with the compound CG165 of the present invention, Stat3 activity (p-STAT3) was inhibited in a concentration-dependent manner, and in particular, Stat3 activity (p-STAT3) was inhibited by more than 50% when treated at 5 μM.

[0159] Example 5: Confirmation of the cancer cell growth inhibitory effect of suppressing Stat 3 phosphorylation in cancer cell lines - 2 In Example 1, it was confirmed that treatment of cancer cells with the compound CG165 of the present invention inhibited Stat 3 activity (p-STAT3) by more than 50%. To confirm the effect of inhibiting Stat 3 activity on the inhibition of cancer cell growth, a cell proliferation assay was performed using a breast cancer cell line, a representative carcinoma.

[0160] Specifically, breast cancer cell line MDA-MB-468 was treated with CG-165 at different concentrations for 24 or 48 hours, and cell viability was measured using the WST-1 (Dojindo Laboratories, Kumamoto, Japan) method.

[0161] As a result, as shown in FIG. 7, it was confirmed that the compound of the present invention, CG165, inhibits the growth of breast cancer cells in a concentration-dependent manner.

[0162] Therefore, it was confirmed that the compound of the present invention, CG165, inhibits the activity of Stat 3 in representative cancer cell lines, such as prostate cancer cell lines and breast cancer cell lines, thereby inhibiting the growth of cancer cells.

[0163] Example 6: Inhibition of Stat3 target protein expression and cell death effects in cancer cell lines In Example 1, it was confirmed that treatment with the compound CG165 of the present invention inhibits Stat3 activity (p-STAT3) in cancer cells. The inhibitory effect of Stat3 target protein expression and the cell death effect due to the inhibition of Stat3 activity were confirmed by Western blotting.

[0164] As a result, as shown in Figure 8, it was confirmed that the compound of the present invention, CG165, significantly suppressed the expression of Cyclin A, Mcl-1, and Survivin, which are target proteins of Stat 3. It was also confirmed that Caspase 3, an enzyme related to cancer cell apoptosis, was activated, which in turn promoted the degradation of PARP protein, a cell death biomarker, confirming that CG165 exerts its anticancer effect through cell death.

[0165] Example 7: Confirmation of the effect of inhibiting Stat 3 phosphorylation in natural killer cells (NKL) The inhibitory effect of CG165 produced in Synthesis Example 3 or CG155 produced in Synthesis Example 6 on Stat3 phosphorylation in natural killer cells (NKL), which are immune cells, was confirmed by Western blotting.

[0166] Specifically, NKL(4x 105 The cells were plated in 6 wells and treated with 2, 5, or 10 μM of CG155 or CG165, respectively, and the level of Stat3 phosphorylation was measured by Western blotting using p-STAT3 antibody.

[0167] As a result, as shown in Figure 9, treatment with the compounds of the present invention, CG155 or CG165, was confirmed to inhibit the level of Stat 3 phosphorylation in immune cells in a concentration-dependent manner, and when treated at 5 uM or higher, both compounds were confirmed to inhibit Stat 3 activation by more than 90%.

[0168] Therefore, it was confirmed that the cinnamamide derivatives of the present invention have the ability to increase the activity of suicide cells.

[0169] Example 8: Confirmation of the inhibitory effect of Stat 3 phosphorylation in bone marrow-derived macrophages (BMDM) Bone marrow-derived macrophages (BMDMs) are cells isolated from bone marrow and have diverse biological functions, such as phagocytosis, which removes pathogens and cellular debris, and inducing immune responses. BMDMs are used to study the functions of these macrophages.

[0170] In response to this, the inhibitory effect of CG165 produced in Synthesis Example 3 or CG155 produced in Synthesis Example 6 on Stat3 phosphorylation in bone marrow-derived macrophages was confirmed by Western blotting.

[0171] Specifically, 10 mL of macrophage culture medium (RPMI1640 containing 10% fetal bovine serum (FBS), 1% penicillin / streptomycin, and 2 mM glutamine) was placed in the bone cavity of a C57BL / 6 mouse, and bone marrow cells were harvested from the medium. Bone marrow-derived macrophages were isolated and incubated from the harvested bone marrow cells using a reported method (J. Weischenfeldt, et al., Cold Spring Harb. Protoc. 3, 5080, 2008).

[0172] To confirm the inhibitory effects of CG155 and CG165 on Stat 3 activation in bone marrow-derived macrophages isolated from mouse bone marrow, bone marrow-derived macrophages were pretreated with DMSO, CG155 (5 μM, 10 μM), or CG165 (2 μM, 5 μM), respectively, for 1 hour. After removing the culture medium, the macrophages were treated with interleukin-10 (IL-10), a cytokine that activates Stat 3, for 15 minutes. The inhibitory effects of CG155 and CG165 on Stat 3 activation were then measured by Western blot analysis using a p-STAT3 antibody.

[0173] As a result, as shown in Figure 10, it was confirmed that treatment with the compounds of the present invention, CG155 or CG165, inhibited the degree of phosphorylation of Stat 3 in macrophages in a concentration-dependent manner, and in the case of CG165, treatment with 5 uM inhibited Stat 3 activation by more than 90%.

[0174] Therefore, it was confirmed that the compounds of the present invention have the ability to regulate not only the activity of suicide cells but also the activity of macrophages.

[0175] Example 9: Confirmation of the growth inhibitory effect on cancer tissue in nude mice The anti-cancer efficacy of oral administration of the compound of the present invention, CG165, was evaluated using a nude mouse subcutaneously transplanted model of human prostate cancer (DU-145).

[0176] Specifically, 5-week-old Athymic-NCr NCr-nu specific pathogen-free (SPF) nude mice were used. The tumor cell concentration was 3 × 10 7 The cell culture medium was adjusted to 100 cells / mL and injected subcutaneously into the axilla between the right shoulder and chest wall at 0.3 mL per mouse. CG165 was then adjusted to the appropriate concentration by adding DMAC 10% + Tween 80 10% + 20% HPβCD 80% in stages and orally administered at 0.2 mL per 20 g mouse for a total of 23 doses over 30 days. Thirty days after the start of drug administration, the DU-145 tumor was excised and weighed.

[0177] As a result, as shown in Figure 11, on the final day (day 30), statistically significant tumor weight reductions of 43.3% (p<0.05) and 64.3% (p<0.05) were confirmed in the CG165 (30 mg / kg) and CG165 (60 mg / kg) groups, respectively, compared to the vehicle control group (vehicle).

[0178] Furthermore, as shown in Figure 12, the results on the final day (day 30) confirmed that, compared to the vehicle control group, the CG165 (PO, 30 mg / kg) and CG165 (60 mg / kg) groups showed statistically significant tumor growth inhibition of 45.6% (p<0.05) and 63.3% (p<0.05), respectively.

[0179] On the other hand, as shown in FIG. 13, it was confirmed that no decrease in body weight was observed in the CG165 (30 mg / kg) or CG165 (60 mg / kg) groups compared to the vehicle control group.

[0180] Therefore, it was confirmed that the compound of the present invention, CG165, effectively inhibits the growth of cancer tissues in animal models.

[0181] Next, the anti-cancer efficacy of the compound of the present invention, CG126, was evaluated by intraperitoneal administration using a nude mouse subcutaneous transplant model of human prostate cancer (DU-145) using the same method as above.

[0182] As a result, as shown in Figure 14, on the final day (day 25), a statistically significant reduction in tumor weight was confirmed at 54.2% (p<0.06) in both the vehicle control group and the CG126 (30 mg / kg) administration group compared to the vehicle control group.

[0183] Therefore, it was confirmed that the compound of the present invention, CG126, effectively inhibits the growth of cancer tissues in animal models.

[0184] Example 10: Verification of single oral administration toxicity in mice In order to obtain information on the acute toxicity of CG165, a compound of the present invention whose anticancer effect has been verified in an animal model, the test substance was orally administered in a single dose of 100, 200, and 300 mg / kg to male mice, and the mortality rate, general symptoms, weight change, and autopsy findings were observed for 7 days.

[0185] Specifically, 20 5-week-old Athymic-NCr NCr-nu specific pathogen-free (SPF) nude mice were used, supplied by Coretec (Pyeongtaek, Gyeonggi-do). CG165 was then orally administered to the mice by adding DMAC 10% + Tween 80 10% + 20% HPβCD 80% in stages to prepare the administration solution for each dose.

[0186] Observations for general symptoms and dead animals were made every hour from 1 hour to 6 hours after the final administration on the day of administration, and once a day from the day after administration until the 7th day, for changes in general symptoms, toxic symptoms, and the presence or absence of dead animals.

[0187] To observe changes in body weight, the body weights of all animals used in the test were measured before the start of administration and on days 0, 1, 2, 4 and 7 after administration.

[0188] On the seventh day after administration, all animals were anesthetized with CO2 gas, and then the abdomen was opened and the abdominal arteries and veins were cut to exsanguinate the animals, after which all internal organs were observed with the naked eye.

[0189] As a result, no animals died, and no general symptoms were observed that were attributable to the administration of the test substance.

[0190] Furthermore, as shown in Figure 15, it was confirmed that there was no statistically significant weight loss after administration until the final day of the test. At the time of autopsy on the final day, no toxicological changes due to the administration of the test substance were observed.

[0191] Therefore, it was confirmed that the compound CG165 of the present invention did not induce toxicological changes when administered orally to male mice at doses of 100, 200, and 300 mg / kg.

[0192] In summary, the novel cinnamamide derivative compounds of the present invention have been confirmed to inhibit Stat 3 activity in cancer cells, suppress the expression of Stat 3 target molecules, and induce cell death. Furthermore, they have been confirmed to inhibit Stat 3 activity in immune cells, such as natural killer cells and macrophages, and thus, the compounds of the present invention have been confirmed to regulate immune cell activity and enhance anti-cancer effects. Furthermore, they have been confirmed to effectively inhibit the growth of cancer tissue in animal models.

[0193] Furthermore, it was confirmed that the compound of the present invention did not induce toxicological changes up to a dose of 300 mg / kg in a single oral administration toxicity study.

[0194] Therefore, it has been confirmed that the novel compound of the present invention or a pharmaceutically acceptable salt thereof exhibits the effect of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by suppressing STAT3 activity, and therefore can be useful for the prevention or treatment of cancer.

[0195] From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are merely illustrative and not limiting. The scope of the present invention should be interpreted as including within the meaning and scope of the claims below, and all modifications and variations derived from the equivalent concepts thereof, rather than the above detailed description.

Claims

1. A compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof: 【Chemical 1】 In the above Chemical Formula 1, R １ is C １－６ Alkyl; Hydroxy; Halogen; C １－６ alkynyloxy; or unsubstituted or substituted C １－６ is alkoxy; The substituted C １－６ Alkoxy is C １－６ containing aryl or 5-14 membered heteroaryl as a substituent; the heteroaryl contains one or more O, N, or S; R ２ is hydrogen or alkyl.

2. A pharmaceutical composition for preventing or treating cancer, comprising, as an active ingredient, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: 【Chemistry 2】 In the above Chemical Formula 1, R １ is C １－６ Alkyl; Hydroxy; Halogen; C １－６ alkynyloxy; or unsubstituted or substituted C １－６ is alkoxy; The substituted C １－６ Alkoxy is C １－６ containing aryl or 5-14 membered heteroaryl as a substituent; the heteroaryl contains one or more O, N, or S; R ２ is hydrogen or alkyl.

3. The composition according to claim 2, wherein the composition inhibits the activity of signal transducers and activators of transcription 3 (STAT3).

4. The composition according to claim 2, characterized in that the composition suppresses the activity of Stat 3 to regulate the activity of immune cells, such as natural killer cells or bone marrow-derived macrophages.

5. 3. The composition of claim 2, wherein the cancer is any one selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, liver cancer, breast cancer, cervical cancer, thyroid cancer, parathyroid cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, ovarian cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, colon cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulva carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, and pituitary adenoma.

6. The composition of claim 2, further comprising a pharmaceutically acceptable carrier, excipient, or diluent.

7. The composition of claim 2 , further comprising an anti-cancer agent.

8. 8. The composition of claim 7, wherein the anticancer agent is at least one selected from the group consisting of DNA alkylating agents, anticancer antibiotics, plant alkaloids, targeted anticancer agents, and cancer immunotherapy.

9. The anticancer drugs include mechlorethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, and dactinomycin (actinomycin). D), doxorubicin (doxorubicin: adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin, C bleomycin (C Bleomycin; vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, irinotecan, Glivec, Tasigna The composition of claim 7, which is any one or more selected from the group consisting of: fluticasone, fluoxetine, fluoxetine-100, fluoxetine-100 agonist ...

10. A method for preventing or treating cancer, comprising administering to an individual other than a human a pharmaceutical composition comprising, as an active ingredient, a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof: 【Chemistry 3】 In the above Chemical Formula 1, R １ is C １－６ Alkyl; Hydroxy; Halogen; C １－６ alkynyloxy; or unsubstituted or substituted C １－６ is alkoxy; The substituted C １－６ Alkoxy is C １－６ containing aryl or 5-14 membered heteroaryl as a substituent; the heteroaryl contains one or more O, N, or S; R ２ is hydrogen or alkyl.

11. The method of claim 10, wherein the composition inhibits the activity of signal transducers and activators of transcription 3 (STAT3).

Images