Novel cinnamamide derivative and use 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 disclosure relates to novel cinnamamide derivatives and their use in preventing or treating cancer. Background Art

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

[0003] Since the first use of chemical anticancer drugs in the 1940s, various anticancer drugs have emerged with the goal of fundamentally eliminating cancer cells and ultimately defeating cancer. The first generation of anticancer drugs developed before the mid-1990s inhibited the growth of rapidly growing cancer cells, but the side effects of affecting normal cells at the same time became a problem. In order to solve this problem, the second generation of anticancer drugs, i.e., targeted anticancer drugs that selectively attack cancer cells, have been developed since the late 1990s, but a problem has been found in which drug resistance will develop after a period of use, thereby reducing the effectiveness of the drug (P. Cohen et al., Nature Rev. Drug Discov. 20, 551, 2021).

[0004] To solve this problem, the third generation of anticancer drugs, namely immune anticancer drugs, have emerged since 2010. Immunotherapy is a treatment method that inhibits or eliminates cancer growth by activating and inhibiting the immune system in the human body, and this method includes immune checkpoint inhibitors, immune cell therapy, therapeutic antibodies, etc. Immune checkpoint inhibitors are drugs that attack cancer cells by activating T cells by blocking the activation of immune checkpoint proteins involved in T cell inhibition. Examples include CTLA-4, PD-1, PD-L1 inhibitors, etc. Immunocell therapy is a drug that enhances cellular immunity against cancer cells by collecting, strengthening and modifying T cells and then returning them to the body through autologous cell transplantation (ACT). Examples include CAR T cell therapy and natural killer cell (NK cell) therapy that induces cancer cell death by inducing innate immune response, etc. In addition, therapeutic antibodies refer to drugs that attack cancer cells by releasing drugs when the antibody-drug complex binds to cancer cells, and examples thereof may include antibody-drug conjugates (ADCs), etc. (Y. Zhang et al., Cell. Mol. Immunol. 17, 807, 2020).

[0005] Cancer immunotherapy is a new type of anticancer drug that connects chemotherapy and targeted therapy, and has achieved clinical success by significantly improving patient survival (ADWaldman et al., Nature Rev.Immunol.20,651,2020). However, the low immune reactivity of cancer cells is considered to be the reason for the decreased response rate of cancer immunotherapy, especially immune checkpoint inhibitors. In particular, the overall reactivity of solid cancers is low (for example, the reactivity of lung cancer is 15% to 20%), and the side effects and drug resistance of cancer immunotherapy have been reported, so its limitations are obvious (G.Morad et al., Cell 184,5309,2021).

[0006] In 2014, Nature published an article titled "Three known unknowns", which pointed out that despite the progress in cancer treatment, there are still three unknowns in cancer: anticancer drug resistance, cancer metastasis and the role of normal cells around cancer cells, namely the tumor microenvironment (K. Bourzac, Nature 509, S69, 2014). It is believed that only by finding the answers to the above three points and developing regulators can humans get rid of cancer.

[0007] The development of anticancer drug resistance can be roughly divided into two types: one is intrinsic resistance, that is, because the resistance factors exist in cancer cells from the beginning, the cancer cells do not respond to the drug; the other is acquired resistance, that is, the drug shows anti-cancer effects for a period of time, but due to mutations or interactions with other signaling systems during continuous administration, the cancer cells no longer respond to the drug.

[0008] There are many reasons for drug resistance, among which the tumor microenvironment has recently become a topic of concern. Tumors themselves have very complex heterogeneity and interact with various cells around them, which are collectively referred to as 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 organically connected to each other and are closely related to anticancer drug resistance (MRJunttila, Nature 501,346,2013). For example, important immune cells in the tumor microenvironment include regulatory T cells (Treg), bone marrow-derived suppressor cells (MDSC), cancer-associated fibroblasts (CAF), natural killer cells (NK cells), etc.

[0009] At the same time, signal transduction and transcription activator protein 3 (STAT3) has been identified as an important protein for intracellular signal transduction, its function is regulated by cytokines or hormones, and it is involved in cell growth and differentiation, etc. Many studies have confirmed that the sustained activity of STAT3 can promote the survival and metastasis of tumor cells (M. El-Tanani et al., Cellular Signalling 92, 100275, 2022). In addition, a number of studies in the past 10 years have found that STAT3 is associated with inducing anticancer drug resistance and maintaining cancer stem cells (P. Shih et al., Drug Discov. Today, 26, 1450, 2021). As mentioned above, STAT3 is involved in regulating the functions of regulatory T cells, bone marrow-derived suppressor cells and natural killer cells in the tumor microenvironment, especially immune cells, which are the main cause of anticancer drug resistance (D. Bayik et al., Nature Rev. Cancer, 21, 526, 2021).

[0010] To this end, it is necessary to conduct research on the development of drugs that overcome anticancer drug resistance and increase drug activity by activating anticancer immunity, which converts the tumor microenvironment from a cold cancer that does not respond to cancer immunotherapy to a hot cancer that responds to cancer immunotherapy.

[0011] Under such circumstances, the present inventors have endeavored to develop a method that can more effectively treat or prevent cancer by controlling the growth of cancer cells and cancer cell microenvironment. Therefore, they synthesized novel cinnamamide derivatives and confirmed that the compounds inhibit the activity of STAT3, thereby showing the effects of inhibiting the growth of cancer cells and enhancing the activity of immune cells that play an important role in the tumor microenvironment, and completed the present disclosure. Summary of the invention

[0012] [Technical issues]

[0013] The present inventors have developed a novel cinnamamide derivative which can be effectively used for preventing or treating cancer, thereby completing the present disclosure.

[0014] [Technical solution]

[0015] One object of the present disclosure is to provide a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0016] [Chemical formula 1]

[0017]

[0018] In Chemical Formula 1,

[0019] R 1 It is C 1-6 Alkyl; Hydroxyl; Halogen; C1-6 Alkynyloxy; or unsubstituted or substituted C 1-6 Alkoxy; wherein substituted C 1-6 Alkoxy contains C 1-6 aryl or 5- to 14-membered heteroaryl as a substituent, wherein the heteroaryl contains one or more O, N or S, and

[0020] R 2 is hydrogen or an alkyl group.

[0021] Another object of the present disclosure is to provide a pharmaceutical composition for preventing or treating cancer, which includes the compound represented by the above Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0022] Still another object of the present disclosure is to provide a method for preventing or treating cancer, comprising administering a pharmaceutical composition containing a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient to a subject other than a human.

[0023] Still another object of the present disclosure is to provide a food composition for preventing or improving cancer, which includes the compound represented by the above Chemical Formula 1 or a sitologically acceptable salt thereof as an active ingredient.

[0024] Even another object of the present disclosure is to provide a feed composition for preventing or ameliorating cancer, which includes the compound represented by the above Chemical Formula 1 or a dietetically acceptable salt thereof as an active ingredient.

[0025] [Beneficial Effects]

[0026] The novel compound or pharmaceutically acceptable salt disclosed in the present invention inhibits the activity of STAT3, thereby inhibiting the growth of cancer cells and enhancing the activity of immune cells in the tumor microenvironment, and thus can be effectively used for the prevention or treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Compound CG165 according to an embodiment of the present disclosure is shown 1 H-NMR data.

[0028] Figure 2 is a schematic diagram showing that the disclosed compound CG165 inhibits STAT3 phosphorylation in a concentration-dependent manner in prostate cancer cell lines.

[0029] Figure 3 is a schematic diagram showing that the disclosed compound CG165 inhibits STAT3 phosphorylation in a time-dependent manner in prostate cancer cell lines.

[0030] Figure 4Results of assaying the cell growth inhibitory activity of the disclosed compound CG165 in prostate cancer cell lines are presented.

[0031] Figure 5 is a schematic diagram showing the results of verifying that the compound CG165 of the present disclosure has no cell growth inhibitory activity in normal cell lines.

[0032] Figure 6 is a schematic diagram showing that the disclosed compound CG165 inhibits STAT3 phosphorylation in breast cancer and lung cancer cell lines in a concentration-dependent manner.

[0033] Figure 7 Results of assaying the cell growth inhibitory activity of the disclosed compound CG165 in breast cancer cell lines are presented.

[0034] Figure 8 is a schematic diagram showing that the compound CG165 of the present disclosure inhibits the expression of STAT3 target protein and induces cell death in cancer cell lines.

[0035] Fig. 9 2 is a schematic diagram showing that the compound CG155 or CG165 of the present disclosure inhibits STAT3 phosphorylation in natural killer cells (NKL) which are immune cells.

[0036] Fig.10 is a schematic diagram showing that the disclosed compound CG155 or CG165 inhibits STAT3 phosphorylation in bone marrow-derived macrophages (BMDMs), which are immune cells.

[0037] Fig.11 is a graph showing the results of measuring changes in cancer tissue weight when the compound CG165 of the present disclosure was orally administered to mice with subcutaneous implantation of prostate cancer.

[0038] Fig.12 is a graph showing the results of measuring changes in the size of cancer tissue when the compound CG165 of the present disclosure was orally administered to mice with subcutaneous implantation of prostate cancer.

[0039] Fig.13 is a graph showing the results of measuring changes in animal body weight when the compound CG165 of the present disclosure was orally administered to mice with subcutaneous implantation of prostate cancer.

[0040] Fig.14 : is a graph showing the results of measuring the change in cancer tissue weight when the compound CG126 of the present disclosure is intraperitoneally administered to mice in which prostate cancer is subcutaneously implanted.

[0041] Fig.15 Schematic diagram showing the results of a single oral dose toxicity validation test of the disclosed compound CG165. DETAILED DESCRIPTION

[0042] The present disclosure will be described in detail below. At the same time, each description and embodiment disclosed herein can be applied to other descriptions and embodiments, respectively. That is to say, all combinations of various elements disclosed herein belong to the scope of the present disclosure. In addition, the scope of the present disclosure is not limited by the specific description described below.

[0043] One aspect of the present disclosure provides a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0044] [Chemical formula 1]

[0045]

[0046] In Chemical Formula 1,

[0047] R 1 It is C 1-6 Alkyl; Hydroxyl; Halogen; C 1-6 Alkynyloxy; or unsubstituted or substituted C 1-6 Alkoxy; wherein substituted C 1-6 Alkoxy contains C 1-6 aryl or 5- to 14-membered heteroaryl as a substituent, wherein the heteroaryl contains one or more O, N or S, and

[0048] R 2 is hydrogen or an alkyl group.

[0049] For example, in the compounds of the present disclosure,

[0050] R 1 Can be -OCH 3 、-CH 3 、-OH、-Cl、 and

[0051] R 2 It may be hydrogen or methyl, but is not limited thereto.

[0052] For example, the compound of the present disclosure may be a compound having the following compound structure.

[0053] [Table 1]

[0054]

[0055]

[0056]

[0057] At the same time, the compounds of the present disclosure may exist in the form of pharmaceutically acceptable salts. Acid addition salts formed from pharmaceutically acceptable free acids may be used as salts. As used herein, the term "pharmaceutically acceptable salt" refers to any organic or inorganic addition salt of a compound, the concentration of which makes it exhibit an effective effect that is relatively non-toxic and harmless to the patient, and the side effects caused by the salt will not impair the beneficial effects of the compound represented by Chemical Formula 1.

[0058] Acid addition salts can be prepared in conventional manner, for example, by dissolving the compound in an excess of aqueous acid and then precipitating the salt with a water-soluble organic solvent such as methanol, ethanol, acetone or acetonitrile. Equimolar amounts of the compound and the acid or alcohol (e.g., ethylene glycol monomethyl ether) in water can be heated and the mixture dried by evaporation, or the precipitated salt can be filtered with suction.

[0059] Here, organic acids and inorganic acids can be used as free acids. As inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. can be used. As organic acids, 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, hydroiodic acid, etc. can be used, but are not limited thereto.

[0060] In addition, a base can be used to prepare a pharmaceutically acceptable metal salt. Alkali metal salts or alkaline earth metal salts are obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. Here, it is preferred to prepare sodium salts, potassium salts or calcium salts as pharmaceutical metal salts, but it is not limited thereto. In addition, a silver salt corresponding to the metal salt can also be obtained by reacting an alkali metal salt or an alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0061] Unless otherwise specifically stated, pharmaceutically acceptable salts of the compounds of the present disclosure may include salts of acidic or basic groups that may be present in the compounds of Chemical Formula 1. For example, pharmaceutically acceptable salts may include sodium, calcium, and potassium salts of hydroxyl groups, and other pharmaceutically acceptable amino salts may include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate), p-toluenesulfonate (toluenesulfonate), etc. In addition, these salts may also be prepared by preparation methods known in the relevant art.

[0062] The salts of the cinnamamide derivative compounds disclosed herein are pharmaceutically acceptable salts, and any cinnamamide derivative salts that exhibit equivalent pharmacological activity to the cinnamamide derivative compounds may be used without limitation.

[0063] In addition, the compound represented by Chemical Formula 1 according to the present disclosure includes not only its pharmaceutically acceptable salts, but also all its solvates or hydrates and all possible stereoisomers that can be prepared therefrom. The solvates and stereoisomers of the compound represented by Chemical Formula 1 can be prepared from the compound represented by Chemical Formula 1 using conventional methods known in the art.

[0064] In addition, the compound represented by Chemical Formula 1 according to the present disclosure can be prepared in a crystalline form or in a non-crystalline form, and when the compound of Chemical Formula 1 is prepared in a crystalline form, it can be optionally hydrated or solvated. In the present disclosure, the compound of Chemical Formula 1 may include not only stoichiometric hydrates but also compounds containing various amounts of water. The solvates of the compounds of Chemical Formula 1 according to the present disclosure include stoichiometric solvates and non-stoichiometric solvates.

[0065] Another aspect of the present disclosure provides a pharmaceutical composition for preventing or treating cancer, comprising the compound represented by the above Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0066] The compound represented by the above Chemical Formula 1, its pharmaceutically acceptable salt, prevention and treatment are as described above.

[0067] As used herein, the term "active ingredient" refers to an ingredient that possesses the desired activity by itself or an ingredient that possesses the desired activity together with an ingredient that is inactive by itself, such as a carrier.

[0068] As used herein, the term "prevention" refers to all actions of inhibiting or delaying the occurrence, spread or recurrence of cancer-related diseases by administering the compositions of the present disclosure, and the term "treatment" refers to all actions of improving or beneficially changing the symptoms of the above-mentioned diseases by administering the compositions of the present disclosure.

[0069] Specifically, the pharmaceutical composition of the present disclosure can prevent or treat cancer-related diseases by inhibiting the activity of signal transducer and activator of transcription 3 (STAT3), but is not limited thereto.

[0070] As used herein, the term "STAT3" is a transcriptional regulator that regulates the expression of genes involved in cell growth, differentiation, and apoptosis through intracellular signaling associated with various cytokines and growth factors. It is known that overexpression and abnormal activation of STAT3 in various cancer cells can lead to malignant cancer cells (HQ Wang et al., MedChem.3, e124, 2022). In addition, it is known that when STAT3 is activated, it regulates the functions of various immune cells such as natural killer cells in the tumor microenvironment, thereby weakening the anti-cancer immune effect (J. Huynh, Nature Rev. Cancer, 19, 82, 2019).

[0071] Specifically, the pharmaceutical composition of the present disclosure can prevent or treat cancer-related diseases by inhibiting the activity of STAT3 and controlling the activity of natural killer cells or bone marrow-derived macrophages, which are immune cells, but is not limited thereto.

[0072] In one embodiment of the present disclosure, it is confirmed that the novel compound of the present disclosure inhibits the phosphorylation of STAT3 in cancer cells, inhibits the growth of cancer cells, inhibits the expression of STAT3 target genes and apoptosis genes, induces apoptosis of cancer cells by decomposing apoptosis marker proteins, and even inhibits the activity of STAT3 in natural killer cells and bone marrow-derived macrophages. The results confirm that the novel compound of the present disclosure exhibits the effect of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by inhibiting the activity of STAT3.

[0073] Specifically, the cancer that can be prevented or treated by the pharmaceutical composition of the present invention can be 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, bile duct 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, rectal cancer, brain tumor, anal cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, vulvar cancer, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic cancer, central nervous system tumors, primary central nervous system lymphoma, spinal cord tumors, brain stem gliomas and pituitary adenomas, but is not limited thereto.

[0074] Specifically, the pharmaceutical composition according to the present disclosure may contain the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and its content is 0.0001% to 99.9% (by weight), specifically 0.01% to 80% (by weight), and more preferably 1% to 10% (by weight), based on the total weight of the composition, but is not limited thereto.

[0075] Compositions of the present disclosure may further include pharmaceutically acceptable carriers, diluents or excipients, and may be prepared into various preparations according to the desired purpose by conventional methods, including oral preparations, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and sterile injectable solutions, etc. In addition, compositions may be used by various routes, including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, etc. Examples of suitable carriers, excipients, and diluents that may be included in the compositions may include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginic acid, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talcum powder, magnesium stearate, and minerals. In addition, compositions of the present disclosure may further include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc.

[0076] Examples of oral solid preparations include tablets, pills, powders, granules, capsules, etc. These solid preparations are prepared by mixing the composition with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate or talc can also be used.

[0077] Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc. Liquid preparations may include various excipients such as wetting agents, sweeteners, aromatics, preservatives, etc. in addition to water, commonly used simple diluents or liquid paraffin.

[0078] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils (such as olive oil) or injectable esters (such as ethyl oleate) may be used. As the matrix of suppository preparations, witepsol, polyethylene glycol, Tween 61, cocoa butter, lauric fat, glycerin gelatin etc. may be used. Meanwhile, injections may include conventional additives such as solubilizers, isotonic agents, suspensions, emulsifiers, stabilizers, preservatives.

[0079] Compositions disclosed herein can be used in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to the amount that is sufficient to treat a disease without causing adverse reactions with a reasonable benefit / risk ratio applicable to any medical method, and the effective dose level can be determined according to the health status of the patient, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the method of application, the time of application, the route of administration and the excretion rate, the duration of treatment, the medicine used simultaneously or in combination, and other factors known to the medical field. Compositions can be used as a single therapeutic agent, can also be used in combination with other therapeutic agents, can be used sequentially, can also be used simultaneously with conventional therapeutic agents, and can also be used once or multiple times. Considering all the above factors, it is important that the dosage of the composition is used, at this dosage, the maximum effect can be achieved with the minimum amount, without producing adverse reactions. Therefore, those skilled in the art can easily determine the dosage of the composition.

[0080] For example, the dosage may be increased or decreased according to the administration route, severity of the disease, gender, body weight, age, etc., and therefore, the above dosage does not limit the scope of the present disclosure in any way.

[0081] Specifically, the pharmaceutical composition of the present disclosure can be administered to animals including humans, for example, at a dosage of 1 mg / kg to 10 mg / kg body weight per day, but is not limited thereto. The frequency of administration of the composition of the present disclosure is not particularly limited, but can be administered once a day or divided into multiple administrations. However, the dosage can be increased or decreased according to 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 disclosure in any way.

[0082] Specifically, the pharmaceutical composition of the present disclosure may further include an anticancer agent, but is not limited thereto.

[0083] More specifically, the anticancer agent may be any one of the group consisting of a DNA alkylating agent, an anticancer antibiotic, a plant alkaloid, a targeted anticancer agent, and a cancer immunotherapy, but is not limited thereto.

[0084] Even more specifically, the anticancer agent can be selected from the group consisting of dichloromethyl diethylamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin C, bleomycin, vincristine, vinblastine, e), paclitaxel, docetaxel, etoposide, teniposide, topotecan, irinotecan, Glivec, Tasigna, Herceptin, Iressa, Nexavar, Yervoy, Opdivo, Keytruda, Atezol, Tecentriq and Bavencio, but not limited thereto.

[0085] There are many reasons for the resistance to anticancer drugs, and the tumor microenvironment has recently become a topic of concern. The tumor itself has a very complex heterogeneity and interacts with various cells around it, which 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 organically linked together and are closely related to the resistance to anticancer drugs (MRJunttila, Nature 501, 346, 2013). For example, important immune cells in the tumor microenvironment include regulatory T cells (Treg), bone marrow-derived suppressor cells (MDSC), cancer-associated fibroblasts (CAF) and natural killer cells (NK cells).

[0086] In this regard, it is known that when STAT3 is activated, it regulates the functions of various immune cells such as natural killer cells in the tumor microenvironment, thereby weakening the anti-cancer immune effect (J. Huynh, Nature Rev. Cancer, 19, 82, 2019).

[0087] Since the novel compound or its pharmaceutically acceptable salt disclosed herein exhibits the effects of inhibiting cancer cell growth and enhancing the activity of immune cells in the tumor microenvironment by inhibiting STAT3 activity, it can be effectively used as a drug that overcomes anticancer drug resistance, enhances drug activity by activating anticancer immunity, and transforms the tumor microenvironment from a cold cancer that is unresponsive to cancer immunotherapy to a hot cancer that is responsive to cancer immunotherapy.

[0088] Yet another aspect of the present disclosure provides a method for preventing or treating cancer, comprising administering to a subject a pharmaceutical composition containing the compound represented by the above Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0089] The compound represented by the above Chemical Formula 1, its pharmaceutically acceptable salt, active ingredient, pharmaceutical composition, cancer, prevention and treatment are as described above.

[0090] As used herein, the term "subject" may refer to all animals that suffer from or may suffer from cancer, such as cows, horses, sheep, pigs, goats, camels, antelopes, dogs, cats, humans, etc., but is not limited thereto.

[0091] The prevention or treatment method of the present disclosure may specifically include the step of administering a pharmaceutically effective amount of the composition to a subject who has or may have cancer.

[0092] As used herein, the term "administration" refers to introducing the composition of the present disclosure into the body of a subject who has or may have symptoms of cancer and related diseases by any suitable method. The composition of the present disclosure can be administered by various routes, such as oral or parenteral administration, as long as it can reach the desired tissue.

[0093] Specifically, the composition of the present disclosure can be administered by intraperitoneal route, intravenous route, intramuscular route, subcutaneous route, intradermal route, oral route, intranasal route, intrapulmonary route, intrarectal route, etc. according to the purpose, but is not particularly limited thereto.

[0094] Specifically, the pharmaceutical composition of the present disclosure can be administered to animals including humans at a dosage of, for example, 1 mg / kg to 10 mg / kg body weight per day, but is not limited thereto. The frequency of administration of the composition of the present disclosure is not particularly limited, but can be administered once a day or divided into multiple administrations. However, the dosage can be increased or decreased according to 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 disclosure in any way.

[0095] Specifically, the pharmaceutical composition can prevent or treat cancer-related diseases by inhibiting the activity of signal transducer and activator of transcription 3 (STAT3), but is not limited thereto.

[0096] Even another aspect of the present disclosure provides a food composition for preventing or improving cancer, comprising the compound represented by the above Chemical Formula 1 or a dietetically acceptable salt thereof as an active ingredient.

[0097] The compound represented by the above Chemical Formula 1, the active ingredient, cancer and prevention are as described above.

[0098] As used herein, the term "sitologically acceptable salt" has the same definition as the term "pharmaceutically acceptable salt."

[0099] As used herein, the term "improvement" may refer to all effects that at least reduce a parameter (eg, degree of symptoms) associated with the disorder to be treated by administering a composition according to the present disclosure.

[0100] As used herein, the term "food" may include all foods in the conventional sense, such as meat, sausage, bread, chocolate, candy, snacks, biscuits, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, multivitamins, health functional foods and health foods, etc.

[0101] The food composition of the present disclosure can be conveniently taken as a daily food, and is therefore expected to have a great cancer improvement effect, and accordingly, can be very effectively used for the purpose of promoting health.

[0102] In addition, the food composition of the present disclosure can also be used as health functional food. As used herein, the term "health functional food (health functional food)" refers to the food prepared and processed using raw materials or ingredients with useful functions for the human body according to the Health Functional Food Act (health functional food law). In particular, the term "functionality (functionality)" refers to the purpose of intake is to control the nutrition required for human body structure and function, or to obtain beneficial health effects, such as physiological effects, etc. The food of the present disclosure can be prepared according to the method commonly used in the art, and raw materials and ingredients commonly used in the art can be added during preparation. In addition, as long as it is a recognized food, its formula is not limited. The food composition of the present disclosure can be prepared into various preparations, and the food composition of the present disclosure is different from general medicines, using food as raw materials, so there is no side effect that may occur during the long-term use of medicines, with strong portability, and can be used as an adjuvant to enhance the anti-cancer effect.

[0103] Health food refers to food that has a positive effect of maintaining or promoting health compared to general food, and health supplementary food refers to food used to supplement health. In some cases, the terms health functional food, health food and health supplementary food can be used interchangeably.

[0104] Specifically, health functional food refers to food prepared by adding the extract of the present disclosure to food raw materials such as beverages, teas, spices, chewing gum and candy, or food prepared by encapsulation, powder or suspension. When health functional food is ingested, a specific health effect can be obtained. However, unlike general drugs, since health functional food uses food as raw materials, it has the advantage that side effects that may be caused by long-term administration of drugs will not occur.

[0105] The food composition may further include a physiologically acceptable carrier, and the type of the carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0106] In addition, the food composition may also contain other ingredients conventionally used in food compositions to improve the sense of smell, taste, vision, etc. For example, the composition may contain vitamins A, C, D, E, B1, B2, B6 and B12, niacin, biotin, folic acid, pantothenic acid, etc. In addition, the food composition may contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) and chromium (Cr). In addition, the food composition may also contain amino acids such as lysine, tryptophan, cysteine, valine, etc.

[0107] In addition, the food composition may also contain food additives, such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, advanced bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar pigments, etc.), color developers (sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (monosodium glutamate (MSG), etc.), sweeteners (gin, sodium cyclamate, saccharin, sodium salt, etc.), flavors (vanillin, lactone, etc.), expanders (alum, potassium hydrogen tartrate, etc.), enhancers, emulsifiers, thickeners (adhesive paste), film formers, gum bases, defoamers, solvents, improvers, etc. Additives can be selected and used in appropriate amounts according to the type of food.

[0108] The food compositions of the present disclosure may include conventional food additives, unless otherwise specified, and eligibility as a "food additive" is determined by standards or guidelines regarding relevant provisions in accordance with the general provisions of the Korean Food Additives Codex and General Test Method and the general test methods approved by the Korean Ministry of Food and Drug Safety (MFDS).

[0109] Items listed in the Korean Food Additives Codex may include, for example, chemical compositions such as ketones, glycine, potassium citrate, niacin, cinnamic acid, natural additives such as persimmon pigment, licorice extract, crystalline cellulose, sorghum pigment, guar gum, and mixed preparations such as sodium L-glutamate agents, alkali agents for noodles, preservative preparations, tar pigment preparations, etc.

[0110] The food composition of the present disclosure may include the compound of Chemical Formula 1 or its dietetic acceptable salt, and its usage is 0.01% to 95% (by weight), preferably 1% to 80% (by weight), based on the total weight of the composition. In addition, the compound of Chemical Formula 1 or its dietetic acceptable salt contained in the food composition of the present disclosure may be obtained by a method identical or similar to the extraction method mentioned when preparing the pharmaceutical composition, but is not limited thereto.

[0111] In addition, the food composition of the present disclosure can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. to achieve the purpose of preventing or improving cancer.

[0112] For example, a health functional food in the form of a tablet can be prepared as follows: a mixture of the compound of Chemical Formula 1 or a dietetically acceptable salt thereof, an excipient, a binder, a disintegrant, and other additives is granulated by a conventional method, and then a compression molding process is performed with a lubricant. Alternatively, the mixture can also be directly subjected to a compression molding process. In addition, if necessary, the health functional food in the form of a tablet can also include a sweetener, and if necessary, a coating material can also be applied.

[0113] In the health functional food in capsule form, hard capsules can be prepared by filling a mixture of additives (such as the compound of Chemical Formula 1 or its dietary acceptable salt and excipients, etc.) or its granules or coated granules into conventional hard capsules, and soft capsules can be prepared by filling a mixture of additives (such as guanine derivatives or its dietary acceptable salt and excipients, etc.) into a capsule matrix (such as gelatin, etc.). If necessary, a plasticizer (such as glycerol or sorbitol, etc.), a colorant, a preservative, etc. can also be added to the soft capsule.

[0114] The health functional food in the form of a pellet can be prepared by molding a mixture of the compound of Chemical Formula 1 or a dietetically acceptable salt thereof, an excipient, a binder and a disintegrant by an appropriate method, and can be coated with sugar or other coating materials, or coated with starch, talcum powder or an appropriate material.

[0115] The health functional food in the form of particles can be produced by making a mixture of the compound of Chemical Formula 1 or its dietetic acceptable salt, excipients, binders and disintegrants into particles by an appropriate method. If necessary, flavoring agents, sweeteners, etc. can also be added. When the particle size of the health functional food in the form of particles is tested with No. 12 (1680 μm), No. 14 (1410 μm) and No. 45 (350 μm) sieves, all particles can pass through the No. 12 sieve, and based on the total amount, the residue of the No. 14 sieve does not exceed 5.0%, and the amount of particles passing the No. 45 sieve does not exceed 15.0%.

[0116] Definitions of terms such as excipients, binders, disintegrants, lubricants, sweeteners, flavoring agents, etc. are described in literature known in the art, and may include terms having the same or similar functions (The Korean Pharmacopoeiareview, Moonsungsa Publishing Co., Ltd., Korea Pharmaceutical University Association, 5th edition, pp. 33-48, 1989).

[0117] Still another aspect of the present disclosure provides a feed composition for preventing or improving cancer, comprising the compound represented by the above Chemical Formula 1 or a dietetically acceptable salt thereof as an active ingredient.

[0118] The compound represented by the above Chemical Formula 1, its dietetically acceptable salt, active ingredient, cancer, and prevention and improvement are as described above.

[0119] As used herein, the term "feed" refers to food ingested by an animal, and specifically, it may refer to materials that provide organic or inorganic nutrients required to maintain animal life or produce meat, milk, etc. The feed may include feed additives and may be prepared in various forms known in the art.

[0120] The composition including the compound of Chemical Formula 1 of the present disclosure or a dietetically acceptable salt thereof can be used to prevent or treat the occurrence of cancer in subjects other than humans (such as livestock or companion animals), and can be used as a functional feed additive or feed composition.

[0121] The amount of the compound of Chemical Formula 1 or a dietary acceptable salt thereof in the feed composition according to the present disclosure can be appropriately adjusted according to the type and age of the livestock to be treated, the application form, the desired effect, etc., for example, in the range of 1% to 99% (by weight), preferably 10% to 90% (by weight), more preferably 20% to 80% (by weight), but is not limited thereto.

[0122] The type of feed is not particularly limited, and feeds conventionally used in the corresponding technical field can be used. Non-limiting examples of feeds include plant feeds, such as grains, root vegetables, food processing byproducts, algae, fiber, oil, starch, cucurbitaceous vegetables or grain byproducts; and animal feeds, such as protein, inorganic substances, oil, minerals, single-cell protein, animal plankton or food. These feeds can be used alone or in combination of at least two feeds.

[0123] In addition, the compounds of the present disclosure or their dietary acceptable salts can be used as feed additives added to feed compositions. Feed additives can be used to improve the productivity or health of target animals, but are not limited thereto. According to the Control of Livestock and Fish Feed Act, the feed additive may correspond to a supplementary feed.

[0124] The feed additive of the present disclosure can be used by further mixing one or more components of organic acids such as citric acid, fumaric acid, adipic acid, lactic acid and natural antioxidants such as polyphenols, catechins, tocopherols, vitamin C, green tea extract, chitosan, tannic acid, etc., and adding other conventional additives such as buffers, antibacterial agents, etc. as needed. In addition, it can be formulated into liquids, capsules, granules or tablets as needed.

[0125] Feed or feed additives may further include substances with various functions, such as supplementing nutrition and preventing weight loss, improving the digestibility and utilization of fiber in feed, improving oil quality, preventing reproductive disorders and improving conception rate, preventing summer high temperature stress, etc. For example, in addition to main ingredients such as amino acids, inorganic salts, vitamins, antioxidants, antifungal and microbial preparations, such as ground or crushed wheat, barley, corn and other plant protein feeds, such as blood meal, meat meal, fish meal and other animal protein feeds, animal fats and vegetable fats, etc., it can also be used together with nutritional supplements, growth promoters, digestion and absorption promoters and preventive agents.

[0126] The feed and feed additives disclosed herein can be used to feed a variety of animals, including mammals and poultry. For example, to maintain or improve the appearance, they can be used for livestock, such as cattle, goats, etc.; and pets, such as dogs and cats, etc., but are not limited thereto.

[0127] Still another aspect of the present disclosure provides use of a composition comprising the compound represented by the above Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient for preventing or treating cancer.

[0128] The compound represented by Chemical Formula 1, its pharmaceutically acceptable salt, active ingredient, cancer, prevention and treatment are as described above.

[0129] [Embodiments of the present invention]

[0130] The present disclosure will be described in detail by way of examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not intended to be limited by these examples.

[0131] Synthesis Example 1: Synthesis of CG133

[0132] In the compound of Chemical Formula 1, CG133 is synthesized by the method shown in the following Reaction Scheme 1:

[0133] [Reaction Scheme 1]

[0134]

[0135] Specifically, methyl 2-hydroxycinnamate (5 g) was dissolved in dichloromethane (200 mL), and then 3,4-dihydropyran (2.6 g, 1.1 equivalents) was added thereto. A small 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, and the solvent was concentrated under reduced pressure and extracted three times with ethyl acetate (EA) and water. Afterwards, the organic layer was heated to MgSO 4 The concentrate was dried, filtered, and concentrated under reduced pressure. The concentrate was used for the next reaction without purification.

[0136] The concentrate and methacrylamide (2.7 g, 1.1 equivalents) were dissolved in DMF (N,N-dimethylformamide, 100 mL), and sodium hydride (800 mg, 1.2 equivalents) was added to the mixed solution, and stirred at room temperature for 7 hours. 50 mL of saturated ammonium chloride solution and water were added to terminate the reaction, and then extracted three times with ethyl acetate and water. Thereafter, the organic layer was concentrated in MgSO 4 The mixture was dried over 400 °C, filtered, and concentrated under reduced pressure. The concentrate was dissolved in 100 mL of methanol, p-toluenesulfonic acid (20 mg) was added thereto, and the mixture was stirred at room temperature for 4 hours. Then, the solvent was concentrated under reduced pressure, and extracted three times with ethyl acetate (EA) and water. Then, the organic layer was concentrated under MgSO 4 The residue was dried on ice, filtered, and concentrated under reduced pressure. The concentrate was purified by column chromatography (EA:hexane=20:80) to give compound CG133 (3.1 g, 47.7%, light yellow powder).

[0137] The mp of compound CG133 is 198°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0138] 1 H-NMR (400MHz, CDCl 3 )δ9.38(brs,1H),9.30(brs,1H),7.85(d,J=16Hz,1H),7.29(d,J=16Hz,1H),7.25(d,J=7.6Hz,1H),6.9 3(t,J=7.6Hz,1H),6.65(d,J=7.6Hz,1H),6.57(t,J=7.6Hz,1H),5.67(s,1H),5.35(s,1H),1.74(s,3H);

[0139] 13 C-NMR (100 MHz, CDCl 3 )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.

[0140] Synthesis Example 2: Synthesis of CG164

[0141] In the compound of Chemical Formula 1, CG164 is synthesized by the method shown in the following Reaction Scheme 2:

[0142] [Reaction Scheme 2]

[0143]

[0144] Specifically, methyl 2-hydroxycinnamate (5 g) was dissolved in acetone (200 mL), and potassium carbonate (4.7 g, 1.2 equivalents) was then added to the solution. Propylene bromide (4.0 g, 1.2 equivalents) was added to the reaction solution, and stirred at 50-60° C. for 6 hours. When the reaction was completed, the reaction solution was stirred at room temperature for 1 hour, filtered, and concentrated under reduced pressure. The concentrate was used for the next reaction without purification.

[0145] The concentrate and methacrylamide (2.7 g, 1.1 equivalents) were dissolved in DMF (N,N-dimethylformamide, 100 mL), and sodium hydride (800 mg, 1.2 equivalents) was added to the mixed solution, and stirred at room temperature for 7 hours. 50 mL of saturated ammonium chloride solution and water were added to terminate the reaction, and then extracted three times with ethyl acetate and water. Thereafter, the organic layer was concentrated in MgSO 4The residue was dried, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography (EA:hexane=15:85) to obtain compound CG164 (3.8 g, 50.4%, white powder).

[0146] Compound CG164 has an mp of 122°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0147] 1 H-NMR (600MHz, CDCl 3 )δ8.32(brs,1H),8.25(d,J=16Hz,1H),7.76(d,J=16Hz,1H),7.71(dd,J=1.8,7.8Hz,1H),7.40(t,J=7.8Hz,1H),7.08(d, J=7.6Hz,1H),7.04(t,J=7.8Hz,1H),5.89(s,1H),5.65(s,1H),4.81(d,J=2.4Hz,2H),2.53(t,J=2.4Hz,1H),2.05(s,3H);

[0148] 13 C-NMR (150 MHz, CDCl 3 )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.

[0149] Synthesis Example 3: Synthesis of CG165

[0150] Among the compounds of Chemical Formula 1, CG165 was synthesized by the following method.

[0151] Specifically, in potassium carbonate (K 2 CO 3 ) in the presence of , CG164 synthesized in Synthesis Example 2 was reacted with methyl iodide in acetone to synthesize a compound named CG165. 1 H-NMR data are shown in Figure 1 shown.

[0152] The mp of compound CG165 is 115°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0153] 1 H-NMR (600MHz, CDCl3 )δ7.87(d,J=15.6Hz,1H),7.47(dd,J=1.8Hz,7.8Hz,1H),7.35(dt,J=1.8Hz,7.8Hz,1H),7.04(d,J=7.8Hz,1H),7.01(t,J=7.8 Hz,1H),6.86(d,J=15.6Hz,1H),5.43(m,2H),4.77(d,J=2.4Hz,2H),3.30(s,3H),2.53(t,J=2.4Hz,1H),2.06(d,J=1.2Hz,3H);

[0154] 13 C-NMR (150 MHz, CDCl 3 )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.

[0155] Synthesis Example 4: Synthesis of CG126

[0156] Among the compounds of Chemical Formula 1, CG126 was synthesized by the following method.

[0157] Specifically, CG126 was synthesized using 2-methylcinnamic acid as the starting material in the same manner as in Reaction Scheme 2 above.

[0158] The mp of compound CG126 is 126°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0159] 1 H-NMR (500MHz, CDCl 3 )δ8.22(d,J=15.5Hz,1H),8.20(brs,1H),7.75(d,J=15.5Hz,1H),7.64(dd,J=1.5Hz,7.5Hz,1H),7.36(dt,J=1.5 Hz,7.7Hz,1H),6.96(t,J=7.5Hz,1H),6.92(d,J=8.0Hz,1H),5.85(s,1H),5.62(m,1H),3.90(s,3H),2.03(s,3H);

[0160] 13 C-NMR (100 MHz, CDCl 3)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.

[0161] Synthesis Example 5: Synthesis of CG127

[0162] Among the compounds of Chemical Formula 1, CG127 was synthesized by the following method.

[0163] Specifically, CG127 was synthesized using 2-methylcinnamic acid as the starting material in the same manner as in Reaction Scheme 2 above.

[0164] The mp of compound CG127 is 135°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0165] 1 H-NMR (500MHz, CDCl 3 )δ8.21(brs,1H),8.18(d,J=16Hz,1H),7.69(d,J=7.5Hz,1H),7.66(d,J=15.5Hz,1H),7.29 (dt,J=1.5Hz,7.5Hz,1H),7.23(m,2H),5.87(s,1H),5.64(m,1H),2.47(s,3H),2.04(s,3H);

[0166] 13 C-NMR (100 MHz, CDCl 3 )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.

[0167] Synthesis Example 6: Synthesis of CG155

[0168] Among the compounds of Chemical Formula 1, CG155 was synthesized by the following method.

[0169] Specifically, CG155 was synthesized by reacting CG133 prepared in Synthesis Example 1 with 4-(bromomethyl)pyridine in the presence of sodium hydride.

[0170] Compound CG155 has an mp of 156°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0171] 1 H-NMR (600MHz, CDCl 3 )δ8.65(s,2H),8.44(brs,1H),8.30(d,J=15.6Hz,1H),7.82(d,J=15.6Hz,1H),7.71(dd,J=1.2Hz,7.8Hz,1H),7.41(d,J=4.8Hz,2H),7. 34(dt,J=1.2Hz,8.4Hz,1H),7.03(t,J=7.8Hz,1H),6.89(d,J=7.8Hz,1H),5.90(s,1H),5.65(d,J=1.2Hz,1H),5.21(s,2H),2.05(s,3H);

[0172] 13 C-NMR (150 MHz, CDCl 3 )δ176.66,166.55,156.97,150.02,145.74,141.18,139.88,131.89,1 29.42,124.13,122.59,121.61,121.56,120.11,112.46,68.63,18.45.

[0173] Synthesis Example 7: Synthesis of CG168

[0174] Among the compounds of Chemical Formula 1, CG168 was synthesized by the following method.

[0175] Specifically, CG168 was synthesized using 2-chlorocinnamic acid as the starting material in the same manner as in Reaction Scheme 2 above.

[0176] The mp of compound CG168 is 147°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0177] 1 H-NMR (600MHz, CDCl 3 )δ8.55(brs,1H),8.29(d,J=16.2Hz,1H),7.79(dd,J=1.2Hz,7.8Hz,1H),7.76(d,J=15.6Hz ,1H),7.44(d,J=7.8Hz,1H),7.31(m,2H),5.94(s,1H),5.69(d,J=1.2Hz,1H),2.06(s,3H);

[0178] 13C-NMR (150 MHz, CDCl 3 )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.

[0179] Synthesis Example 8: Synthesis of CG187

[0180] Among the compounds of Chemical Formula 1, CG187 was synthesized by the following method.

[0181] Specifically, CG187 was synthesized using 2-hydroxycinnamic acid as the starting material in the same manner as in Reaction Scheme 2 above.

[0182] The mp of compound CG187 is 127°C, and 1 H- and 13 The results of C-NMR analysis are as follows:

[0183] 1 H-NMR (600MHz, CDCl 3 )δ8.43(brs,1H),8.31(d,J=15.6Hz,1H),7.76(d,J=16.2Hz,1H),7.70(dd,J=1.2Hz,7.8Hz,1H ),7.5-7.3(m,6H),6.98(m,2H),5.89(s,1H),5.62(d,J=1.2Hz,1H),5.20(s,2H),2.04(s,3H);

[0184] 13 C-NMR (100 MHz, CDCl 3 )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.

[0185] Experimental Example 1: Cell culture

[0186] 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, Virginia, USA). Prostate cancer cell line DU-145, breast cancer cell line MDA-MB-468, lung cancer cell line HCC827 and human foreskin fibroblast HFF were cultured in RPMI-1640 (Gibco, Gaithersburg, Maryland, USA) medium supplemented with 10% FBS (Gibco) and 100 U / mL penicillin. Human breast epithelial cells MCF10A were cultured in DMEM-F12 (Gibco) medium supplemented with 10% fetal bovine serum (FBS), 0.5% penicillin / streptomycin, EGF (20 ng / mL), insulin (10 μg / mL) and hydrocortisone (0.5 mg / mL). The natural killer cell NKL cell line was cultured 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 cultured in 5% CO 2 Culture in an incubator.

[0187] Experimental Example 2: Western blot analysis

[0188] Prostate cancer cell line DU-145, breast cancer cell line MDA-MB 468, lung cancer cell line HCC827, natural killer cells (NKL) or bone marrow-derived macrophages (BMDM) (4×10 5These immune cells were cultured in 6-well plates. After 24 hours, the cells were treated with 0.1% DMSO, HCA, EA and the compounds prepared in Synthesis Examples 1 to 8 according to the concentration and time, rinsed with PBS, and recovered with RIPA buffer. 20 g of protein quantified with Bio-Rad protein detection kit (Bio-Rad, Hercules, California, USA) was subjected to 8% SDS-PAGE electrophoresis and then transferred to a PVDF membrane (EMDMillipore, Billerica, Massachusetts, USA). After blocking the membrane with TBST containing 5% skim milk, the primary and secondary antibodies were used for the experiment. 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) and PARP (Cell Signaling) GAPDH (Santa Cruz), and the secondary antibodies used were HRP-conjugated goat anti-rabbit and anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, Pennsylvania, USA). All antibodies used were washed three times with TBST, and the bands were detected using Luminata Forte Western HRP substrate (EMD Millipore) and LAS 4000mini (GE Healthcare Life Sciences). Subsequently, GAPDH was calibrated using the MultiGauge program (Fuji Photo Film, Tokyo, Japan).

[0189] Example 1: Confirmation of the STAT3 phosphorylation inhibitory effect in cancer cell line-1

[0190] The STAT3 phosphorylation inhibitory effects of the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8 in cancer cell lines were confirmed by Western blotting.

[0191] Specifically, rectal cancer cells were used as a representative example of cancer, and the STAT3 phosphorylation inhibitory effect was confirmed therein. The results are shown in Table 2 below.

[0192] [Table 2]

[0193]

[0194]

[0195] The results confirmed that the cinnamamide derivative compounds of Chemical Formula 1 prepared in Synthesis Examples 1 to 8 of the present disclosure could inhibit STAT3 activity (p-STAT3) by more than 50%.

[0196] Next, the STAT3 phosphorylation inhibitory effect of the compound CG165 of the present disclosure in cancer cell lines was confirmed by Western blotting.

[0197] Results, such as Figure 2 and Figure 3 As shown, it was confirmed that in the prostate cancer cell line DU-145, treatment with the disclosed compound CG165 inhibited STAT3 activity (p-STAT3) in a concentration-dependent manner. In particular, it was confirmed that when the treatment concentration was 2 μM, STAT3 activity (p-STAT3) was inhibited by more than 50% ( Figure 2 ).

[0198] Furthermore, it was demonstrated that treatment with the disclosed compound CG165 inhibited STAT3 activity (p-STAT3) in a time-dependent manner in the prostate cancer cell line DU-145 ( Figure 3 ).

[0199] Example 2: Demonstration of the effect of inhibiting STAT3 phosphorylation in cancer cell line-1 on cancer cell growth inhibition

[0200] In Example 1, it was confirmed that STAT3 activity (p-STAT3) was inhibited when cancer cells were treated with the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8. In order to confirm the effect of inhibiting STAT3 activity on inhibiting the growth of cancer cells, a cell proliferation assay was performed using a representative cancer cell, a prostate cancer cell line.

[0201] Specifically, the prostate cancer cell line DU-145 (0.8×10 4 Cells were cultured in 96-well plates. After 24 hours, DMSO (control) and CG-165 were treated for 24 or 48 hours according to the concentration, and then treated with 10ul of WST-1 reagent (Dojindo Laboratories, Kumamoto, Japan). After about 1 hour of treatment, the cell viability was calculated by the absorbance measured at 450nm using a microplate reader (Multiskan GO system, Thermo Fisher Scientific, Rockford, IL, USA).

[0202] Results, such as Figure 4 As shown, it was confirmed that the compound CG165 of the present disclosure inhibited the growth of cancer cells in a concentration-dependent manner.

[0203] Therefore, it was confirmed that the compound CG165 of the present disclosure inhibits the growth of cancer cells by inhibiting the activity of STAT3 in cancer cells.

[0204] Example 3: Confirmation of the effects of the disclosed compounds on normal cells

[0205] To confirm that the disclosed compound CG165 does not affect the growth of normal cells, human foreskin fibroblasts HFF and human mammary epithelial cells MCF10A were treated with CG-165 according to the concentration for 48 hours, and then the cell viability was determined using the WST-1 assay (Dojindo Laboratories, Kumamoto, Japan).

[0206] Results, such as Figure 5 As shown, it was confirmed that the compound CG165 of the present disclosure had no significant effect on the growth of normal cells even when treated at a concentration of 10 μM for 48 hours.

[0207] Therefore, it was confirmed that the compound CG165 of the present disclosure had almost no effect on normal cells and selectively inhibited the growth of cancer cells.

[0208] Example 4: Confirmation of the STAT3 phosphorylation inhibitory effect in cancer cell line-2

[0209] Taking representative cancers other than prostate cancer cell lines as examples, the STAT3 phosphorylation inhibitory effects of the cinnamamide derivatives of Chemical Formula 1 prepared in Synthesis Examples 1 to 8 were confirmed in breast cancer cell lines and lung cancer cell lines by Western blotting.

[0210] Specifically, the STAT3 phosphorylation inhibitory effect of CG165 of the present disclosure in breast cancer cell line MDA-MB-468 and lung cancer cell line HCC827 was confirmed by Western blotting.

[0211] Results, such as Figure 6 As shown, it was confirmed that when the breast cancer cell line MDA-MB-468 and the lung cancer cell line HCC827 were treated with the compound CG165 disclosed in the present invention, STAT3 activity (p-STAT3) was inhibited in a concentration-dependent manner. In particular, it was confirmed that when the treatment concentration was 5 μM, STAT3 activity (p-STAT3) was inhibited by more than 50%.

[0212] Example 5: Confirmation of the inhibitory effect of inhibiting STAT3 phosphorylation on cancer cell growth in cancer cell line-2

[0213] In Example 1, it was confirmed that when cancer cells were treated with the compound CG165 disclosed in the present invention, STAT3 activity (p-STAT3) was inhibited by more than 50%. In order to confirm the effect of inhibiting STAT3 activity on inhibiting cancer cell growth, a cell proliferation assay was performed using a representative cancer, breast cancer cell line.

[0214] Specifically, breast cancer cell line MDA-MB-468 was treated with CG-165 for 24 or 48 h, depending on the concentration, and then cell viability was determined using the WST-1 assay (Dojindo Laboratories, Kumamoto, Japan).

[0215] Results, such as Figure 7 As shown, it was confirmed that the compound CG165 of the present disclosure inhibited the growth of breast cancer cells in a concentration-dependent manner.

[0216] Therefore, it was confirmed that the compound CG165 of the present disclosure inhibited the growth of cancer cells in representative cancers, prostate cancer cell line and breast cancer cell line, by inhibiting the activity of STAT3.

[0217] Example 6: Demonstration of the expression inhibition and apoptotic effect of STAT3 target protein in cancer cell lines

[0218] In Example 1, it was demonstrated that when cancer cells were treated with the compound CG165 of the present disclosure, STAT3 activity (p-STAT3) was inhibited. The effect of inhibiting the expression of STAT3 target protein and the apoptotic effect caused by the inhibition of STAT3 activity were demonstrated using Western blotting.

[0219] Results, such as Figure 8 As shown, it was confirmed that the compound CG165 of the present disclosure significantly inhibited the expression of cyclin A, Mcl-1 and survivin, which are target proteins of STAT3. In addition, it was also confirmed that caspase 3, an enzyme associated with cancer cell death, was activated, thereby promoting the decomposition of PARP protein, a cell death biomarker, thereby confirming that CG165 exerts an anti-cancer effect through cell death.

[0220] Example 7: Demonstration of the inhibitory effect of STAT3 phosphorylation in natural killer cells (NKL)

[0221] The STAT3 phosphorylation inhibitory effect of CG165 prepared in Synthesis Example 3 or CG155 prepared in Synthesis Example 6 in immune cell natural killer cells (NKL) was confirmed by Western blotting.

[0222] Specifically, NKL (4×10 5Cells (100 cells) were cultured in 6-well plates and treated with 2 μM, 5 μM, and 10 μM of CG155 or CG165, respectively, and then the phosphorylation level of STAT3 was determined by Western blotting using a p-STAT3 antibody.

[0223] Results, such as Fig. 9 As shown, it was confirmed that the phosphorylation degree of STAT3 in immune cells was inhibited in a concentration-dependent manner when treated with the compounds CG155 or CG165 of the present disclosure. In addition, when treated at a concentration of 5 μM or higher, the inhibition rate of STAT3 activation by these two compounds exceeded 90%.

[0224] Therefore, it was confirmed that the cinnamamide derivatives disclosed in the present invention have the ability to enhance the suicide cell activity.

[0225] Example 8: Confirmation of the inhibitory effect of STAT3 phosphorylation in bone marrow-derived macrophages (BMDM)

[0226] Bone marrow-derived macrophages (BMDM) are cells isolated from the bone marrow and have a variety of biological functions, such as phagocytosis to remove pathogens and cell debris, inducing immune responses, etc. To study the functions of macrophages, bone marrow-derived macrophages were used.

[0227] Therefore, the STAT3 phosphorylation inhibitory effect of CG165 prepared in Synthesis Example 3 or CG155 prepared in Synthesis Example 6 in bone marrow-derived macrophages was confirmed by Western blotting.

[0228] Specifically, 10 mL of RPMI1640 (a macrophage culture medium) containing 10% fetal bovine serum (FBS), 1% penicillin / streptomycin and 2 mM glutamine was added to the bone cavity between the bones of C57BL / 6 mice, and bone marrow cells were collected therefrom. Bone marrow-derived macrophages were isolated and cultured from the collected bone marrow cells using a reported method (J. Weischenfeldt et al., Cold Spring Harb. Protoc. 3, 5080, 2008).

[0229] Subsequently, to confirm the function of CG155 and CG165 in inhibiting STAT3 activation in bone marrow-derived macrophages isolated from mouse bone marrow, bone marrow-derived macrophages were pretreated with DMSO, CG155 (5 μM, 10 μM), and CG165 (2 μM, 5 μM), respectively, for 1 hour. After removing the culture medium, bone marrow-derived macrophages were treated with interleukin-10 (IL-10), a cytokine that can activate STAT3, for 15 minutes, and then the inhibitory effects of CG155 and CG165 on STAT3 activation were determined by Western blotting using a p-STAT3 antibody.

[0230] Results, such as Fig.10 As shown, it was confirmed that the phosphorylation degree of STAT3 in macrophages was inhibited in a concentration-dependent manner when treated with the disclosed compound CG155 or CG165. In the case of CG165, when treated at a concentration of 5 μM, it was confirmed that the activation of STAT3 was inhibited by more than 90%.

[0231] Therefore, it was demonstrated that the compounds of the present disclosure have the ability to modulate the activity of macrophages and suicide cells.

[0232] Example 9: Confirmation of the Growth Inhibitory Effect on Cancer Tissue in Nude Mice

[0233] An attempt was made to evaluate the anticancer efficacy of the disclosed compound CG165 by oral administration using a human prostate cancer (DU-145) nude mouse subcutaneous transplantation model.

[0234] Specifically, 5-week-old specific pathogen-free (SPF) nude mice Athymic-NCr NCr-nu provided by KOATECH (Pyeongtaek-si, Gyeonggi-do) were used as animals. The cancer cell concentration was adjusted to 3 × 10 7 Cells / mL, and then 0.3mL of cell culture was subcutaneously injected into the axillary area between the right shoulder and chest wall for each mouse. Subsequently, the corresponding concentration of CG165 was prepared by gradually adding DMAC 10% + Tween 80 10% + 20% HPβCD 80%, and orally administered at a dose of 0.2mL per 20g mouse, repeated 23 times for 30 days. 30 days after the start of drug administration, DU-145 tumors were removed and their weights were measured.

[0235] Results, such as Fig.11 As shown, as of the last day (Day 30), it was confirmed that the tumor weight of the CG165 (30 mg / kg)- or CG165 (60 mg / kg)-administered group was statistically significantly reduced by 43.3% (p<0.05) and 64.3% (p<0.05), respectively, compared with the vehicle.

[0236] In addition, if Fig.12 As shown, the results on the last day (day 30) showed that the statistically significant tumor growth inhibition rates of the CG165 (PO, 30 mg / kg)- or CG165 (60 mg / kg)-administered groups compared with the vehicle were 45.6% (p<0.05) and 63.3% (p<0.05), respectively.

[0237] At the same time, if Fig.13As shown, it was confirmed that the body weight of the CG165 (30 mg / kg)- or CG165 (60 mg / kg)-administered group did not decrease compared with the vehicle.

[0238] Therefore, it was confirmed that the compound CG165 of the present disclosure effectively inhibited the growth of cancer tissue in animal models.

[0239] Next, the anticancer efficacy of the compound CG126 of the present disclosure was evaluated in the same manner as above by intraperitoneal administration using a nude mouse subcutaneously transplanted human prostate cancer (DU-145) model.

[0240] Results, such as Fig.14 As shown, as of the last day (day 25), the CG126 (30 mg / kg)-administered group had a statistically significant tumor weight reduction of 54.2% compared with the vehicle (P<0.06).

[0241] Therefore, it was confirmed that the compound CG126 of the present disclosure effectively inhibited the growth of cancer tissue in animal models.

[0242] Example 10: Verification of single oral dose toxicity in mice

[0243] In order to obtain acute toxicity information of the disclosed compound CG165 (whose anticancer effect has been verified in animal models), the test substance was orally administered once to male mice at doses of 100 mg / kg, 200 mg / kg and 300 mg / kg to determine mortality, systemic symptoms, body weight changes and postmortem conditions within 7 days.

[0244] Specifically, the animals were 20 5-week-old specific pathogen-free (SPF) nude mice Athymic-NCr NCr-nu provided by KOATECH (Pyeongtaek-si, Gyeonggi-do). Then, different concentrations of CG165 were prepared by gradually adding DMAC 10% + Tween 80 10% + 20% HPβCD 80% as an administration solution and orally administered.

[0245] Observe the general symptoms and animal deaths every hour within 1 to 6 hours after the last application on the day of application, and observe once a day from the second to the seventh day after application to observe changes in general symptoms, poisoning symptoms and animal deaths.

[0246] In order to observe the changes in body weight, the body weights of all experimental animals were measured before administration and on days 0, 1, 2, 4, and 7 after administration.

[0247] On the 7th day after administration, all animals were anesthetized with carbon dioxide gas, laparotomy was performed, and the animals were bled to death after cutting off the abdominal arteries and veins, and all internal organs were observed with the naked eye.

[0248] As a result, no animal death was observed, and no systemic symptoms due to the administration of the test substance were observed.

[0249] In addition, if Fig.15 The results show that although changes in body weight were observed, there was no statistically significant decrease in body weight until the last day of the test after administration. No toxicological changes due to the administration of the test substance were observed in the autopsy on the last day.

[0250] Therefore, it was confirmed that the compound CG165 of the present disclosure did not cause toxicological changes when administered orally in male mice at a single dose of 100 mg / kg, 200 mg / kg, and 300 mg / kg.

[0251] In summary, it has been confirmed that the novel cinnamamide derivative compounds disclosed herein inhibit the activity of STAT3 in cancer cells, inhibit the expression of STAT3 target molecules, and induce cell death. In addition, the inhibition of STAT3 activity has also been confirmed in natural killer cells and macrophages as immune cells, and thus it has been confirmed that the compounds disclosed herein can improve the anti-cancer effect by regulating the activity of immune cells. In addition, it has also been confirmed that the compounds disclosed herein effectively inhibit the growth of cancer tissues in animal models.

[0252] Furthermore, in a single oral dose toxicity test, it was confirmed that the compounds of the present disclosure did not cause toxic changes until the dose reached 300 mg / kg.

[0253] Therefore, the novel compound or a pharmaceutically acceptable salt thereof disclosed in the present invention exhibits the effects of inhibiting cancer cell growth and enhancing the activity of immune cells in the tumor microenvironment by inhibiting the activity of STAT3, and thus can be effectively used for the prevention or treatment of cancer.

[0254] Those of ordinary skill in the art will recognize that the present disclosure may be implemented in other specific forms without departing from the spirit or essential features of the present disclosure. The described embodiments should be considered in all respects as illustrative only and not restrictive. Therefore, the scope of the present disclosure is described by the appended claims rather than the preceding description. All changes within the meaning and equivalent range of the claims should be included within the scope of the present disclosure.

Claims

1. A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical formula 1] In Chemical Formula 1, R1 is C 1-6 Alkyl; Hydroxyl; Halogen; C 1-6 Alkynyloxy; or unsubstituted or substituted C 1-6 Alkoxy; wherein the substituted C 1-6 Alkoxy contains C 1-6 aryl or 5- to 14-membered heteroaryl as a substituent, wherein the heteroaryl contains one or more O, N or S, and R2 is hydrogen or alkyl.

2. A pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical formula 1] In Chemical Formula 1, R1 is C 1-6 Alkyl; Hydroxyl; Halogen; C 1-6 Alkynyloxy; or unsubstituted or substituted C 1-6 Alkoxy; wherein the substituted C 1-6 Alkoxy contains C 1-6 aryl or 5- to 14-membered heteroaryl as a substituent, wherein the heteroaryl contains one or more O, N or S, and R2 is hydrogen or alkyl.

3. The composition according to claim 2, wherein the composition inhibits the activity of signal transducer and activator of transcription 3 (STAT3). 4 . The composition according to claim 2 , wherein the composition regulates the activity of natural killer cells or bone marrow-derived macrophages as immune cells by inhibiting the activity of STAT3.

5. 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, bile duct 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, rectal cancer, brain tumor, anal cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, vulvar cancer, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvis cancer, central nervous system tumors, primary central nervous system lymphoma, spinal cord tumors, brain stem gliomas, and pituitary adenomas.

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

7. The composition of claim 2, wherein the composition further comprises an anticancer agent.

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

9. The composition according to claim 7, wherein the anticancer agent is any one selected from the group consisting of dichloromethyl diethylamine, chlorambucil, phenylalanine, nitrogen mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin C, bleomycin, vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, irinotecan, Gleevec, Dasina, Herceptin, Iressa, Nexavar, Y drug, Opdivo, Keytruda, Atezolizumab, Taishengqi and Bavincia.

10. A method for preventing or treating cancer, comprising administering to a subject other than a human a pharmaceutical composition containing a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical formula 1] In Chemical Formula 1, R1 is C 1-6 Alkyl; Hydroxyl; Halogen; C 1-6 Alkynyloxy; or unsubstituted or substituted C 1-6 Alkoxy; wherein the substituted C 1-6 Alkoxy contains C 1-6 aryl or 5- to 14-membered heteroaryl as a substituent, wherein the heteroaryl contains one or more O, N or S, and R2 is hydrogen or alkyl.

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

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