KR102665346B1 - Novel Dichlorophenoxyacetic acid derivatives and use thereof - Google Patents

Abstract

The present invention relates to novel dichlorophenoxyacetic acid derivatives and their use in preventing or treating cancer.

Description

Novel Dichlorophenoxyacetic acid derivatives and use thereof}

The present invention relates to novel dichlorophenoxyacetic acid derivatives and their use in preventing or treating cancer.

According to statistics released in May 2021 (Global Cancer Statistics 2020), 19.3 million new cancer patients are occurring worldwide as of 2020, and the number of deaths due to cancer reaches 10 million (H. Sung et al., CA Cancer J. Clin. 71, 209, 2021).

In this regard, various anticancer drugs have been developed with the goal of eliminating cancer cells at the source and ultimately conquering cancer. The first-generation anticancer drugs developed until the mid-1990s were drugs that inhibited the division of rapidly growing cancer cells, but had the problem of side effects that also affected normal cells. To solve this problem, second-generation anticancer drugs, or targeted anticancer drugs, that selectively attack cancer cells have been developed since the late 1990s, but there is a problem in that resistance develops after a certain period of time and the effectiveness of the drug decreases (P. Cohen et al. Nature Rev Drug Disco 20, 551, 2021).

To solve this problem, third-generation anticancer drugs, or immune anticancer drugs, are being studied in the 2010s. Specifically, a protein called PD-L1 is expressed in cancer cells, and PD-1 or CTLA-4 is produced in immune cells on the other side. When substances from these cells bind, the immune cells are unable to recognize the cancer cells. Therefore, immune checkpoint inhibition, which blocks the binding between PD-L1, PD-1, and CRLA-4, allows T cells to accurately recognize cancer cells and suppress cancer cells (Y. Yang, J Clin Invest. 125, 3335, 2015).

However, in the case of the above immuno-anticancer drugs, the overall reactivity is low in most solid tumors (for example, 15 to 20% in the case of lung cancer), and side effects and resistance to immuno-anticancer drugs have been reported, so the limitations are clear (P. Sharma, et al., Cell 168, 707, 2017).

Meanwhile, the occurrence of anticancer drug resistance is largely divided into two types: intrinsic resistance, in which cancer cells do not respond to the drug due to factors that are resistant to the drug from the beginning, and persistent anticancer effect due to the drug. Acquired resistance occurs when a drug no longer responds due to mutations occurring during the administration process or interactions with other signaling systems.

There are many causes of resistance, and among them, the tumor microenvironment has recently become the subject of interest. Tumors themselves have very complex heterogeneity and interact with a wide variety of cells around them. This is collectively called 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 connected to each other and are closely related to anticancer drug resistance (M.R. Junttila, Nature 501, 346, 2013). For example, important immune cells in the tumor microenvironment include regulatory T cells (Treg), myeloid-derived suppressor cells (MDSC), fibroblasts (cancer-associated fibroblasts (CAF)), and natural killers. There are cells (NK cells, Natural killer cells), etc.

In this regard, the development of drugs that overcome the resistance of anticancer drugs and increase their activity by activating anticancer immunity, which changes the tumor microenvironment from a cancer that does not respond to immunotherapy drugs (cold tumor) to a cancer that responds to immunotherapy drugs (hot tumor). The need for research is emerging.

Under this background, the present inventors have made diligent efforts to develop a method that can more effectively treat or prevent cancer by controlling the growth of cancer cells and the cancer cell microenvironment. As a result, they have synthesized a novel dichlorophenoxyacetic acid derivative and discovered that the compound activates STAT3 By inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment, the present invention was completed.

One object of the present invention is to provide a compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof:

[Formula 1]

.

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

Another object of the present invention is to prevent cancer, comprising the step of administering a pharmaceutical composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient to an entity other than a human. It provides a treatment method.

Another object of the present invention is to provide a food composition for preventing or improving cancer, comprising the compound represented by Formula 1 above, or a foodologically acceptable salt thereof, as an active ingredient.

Another object of the present invention is to provide a feed composition for preventing or improving cancer, comprising the compound represented by Formula 1 above, or a foodologically acceptable salt thereof, as an active ingredient.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Additionally, the scope of the present invention cannot be considered limited by the specific description described below.

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

[Formula 1]

.

For example, the compound is [(E)-2-(3-oxoprop-1-en-1-yl)phenyl] 2-[2,3-dichloro-4-(2-methylidenebutanoyl)phenoxy ] Acetate ([(E)-2-(3-oxoprop-1-en-1-yl)phenyl] 2-[2,3-dichloro-4-(2-methylidenebutanoyl)phenoxy]acetate).

The compounds of the present invention may exist in the form of pharmaceutically acceptable salts. As a salt, an acid salt formed by a pharmaceutically acceptable free acid is useful. As used herein, the term "pharmaceutically acceptable salt" refers to a concentration of a compound having an effective effect that is relatively non-toxic and harmless to patients, and side effects due to the salt do not reduce the beneficial efficacy of the compound represented by Formula 1. means any and all organic or inorganic addition salts.

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

At this time, organic acids and inorganic acids can be used as free acids. Hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. can be used as inorganic acids, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, and maleic acid can be used as organic acids. (maleic acid), succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic 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 is not limited to these.

Additionally, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal salt or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare metal salts, especially sodium, potassium, or calcium salts, but is not limited to these. Additionally, the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (eg, silver nitrate).

Pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compound of Formula 1 above, unless otherwise indicated. 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, and hydrogen phosphate. , dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, etc., and the preparation of salts known in the art. It can be manufactured through a method.

The salt of the dichlorophenoxyacetic acid derivative compound of the present invention is a pharmaceutically acceptable salt, and any salt of the dichlorophenoxyacetic acid derivative that exhibits pharmacological activity equivalent to that of the dichlorophenoxyacetic acid derivative compound can be used without limitation.

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

Furthermore, the compound represented by Formula 1 according to the present invention can be prepared in crystalline form or amorphous form, and when prepared in crystalline form, it can be arbitrarily hydrated or solvated. In the present invention, not only stoichiometric hydrates of the compound represented by Formula 1 above, but also compounds containing various amounts of water may be included. Solvates of the compound represented by Formula 1 according to the present invention include both stoichiometric solvates and non-stoichiometric solvates.

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

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

The term "active ingredient" in the present invention refers to an ingredient that can exhibit the desired activity alone or in combination with a carrier that is inactive on its own.

The term "prevention" of the present invention refers to all actions that suppress or delay the occurrence, spread, and recurrence of cancer-related diseases by administration of the composition of the present invention, and the term "treatment" refers to the treatment of said disease by administration of the composition of the present invention. It refers to any action that improves or changes symptoms to a benefit.

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.

The term “STAT 3” of the present invention is a transcriptional regulatory factor that regulates the expression of genes related to cell growth, differentiation, and death by transmitting signals corresponding to various cytokines and growth factors from outside the cell into the cell. means. Stat 3 is known to contribute to the malignancy of cancer cells through overexpression and abnormal activation in various cancer cells (Kim, et al Arch. Pharm. Res. 39, 1085, 2016). (Kim, et al Arch. Pharm. Res. 39, 1085, 2016). In addition, it is known that when STAT 3 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 (H. Yu, Nature Reviews Immunology 7, 41, 2007).

In one embodiment of the present invention, the novel compound of the present invention inhibits the phosphorylation of Stat 3 in cancer cells, inhibits the growth of cancer cells, inhibits the expression of Stat 3 target genes and apoptosis genes, and induces cell death. It was confirmed that it induces cancer cell death by decomposing marker proteins and inhibits the activity of STAT 3 in natural killer cells. As a result, the novel compound of the present invention inhibits the growth of cancer cells and inhibits the growth of cancer cells in the tumor microenvironment by inhibiting STAT3 activity. It was confirmed that it showed an effect of increasing the activity of immune cells.

Specifically, cancers that can be prevented or treated with the pharmaceutical composition of the present invention include colon cancer, pancreatic cancer, stomach 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, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, Vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine 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, brainstem glioma, and It may be any one selected from the group consisting of pituitary adenoma, but is not limited thereto.

Specifically, the pharmaceutical composition according to the present invention contains a compound represented by Chemical Formula 1, a tautomer thereof, a stereoisomer thereof, a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, and the total weight of the final composition is As a standard, it may be included in an amount of 0.0001 to 99.9% by weight, specifically 0.01 to 80% by weight, and more specifically 1 to 10% by weight, but is not limited thereto.

The composition of the present invention may further include a pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc. according to conventional methods to suit each purpose of use. It can be formulated and used in a variety of forms, such as oral formulations such as aerosols and injections of sterile injectable solutions, and can be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration. Examples of suitable carriers, excipients or diluents that may be included in such compositions include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, Examples include cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, the composition of the present invention may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. Formulated by mixing. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used.

Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. You can.

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable esters such as ethyl oleate. The basis of suppositories is Wethepsol, Macrogol, and Twin 61. Cacao, laurel, glycerogeratin, etc. can be used. Meanwhile, injectables may contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, etc.

The composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" of the present invention refers to an amount that is sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment and does not cause side effects, and the effective dose level refers to the patient's health condition, Factors including the type of disease, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, drugs combined or used simultaneously, and other factors well known in the field of medicine. You can. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

For example, the dosage may increase or decrease depending on the route of administration, severity of disease, gender, weight, age, etc., so the above dosage does not limit the scope of the present invention in any way.

Specifically, the dosage of the pharmaceutical composition of the present invention may be, for example, 0.1 to 500 mg/kg of 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, but may be administered once a day or divided into multiple doses. However, since it may increase or decrease depending on the route of administration, severity of disease, gender, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

Specifically, the composition may additionally include an anticancer agent, but is not limited thereto.

More specifically, the anti-cancer agent is from the group consisting of DNA alkylating agents, anti-cancer antibiotics, plant alkaloids, targeted anti-cancer agents, and cancer immunotherapy. It may be one or more selected items, but is not limited thereto.

More specifically, the anticancer agents include mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, and carmustine. (carmustine: BCNU), lomustine (lomustine: CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin : actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin, C bre C Bleomycin; Vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, iridotecan, glivec ), tasigna, Herceptin, iressa, nexsavar, yervoy, opdovo, keytruda, Atezol, tecentriq ) It may be one or more selected from the group consisting of alc and babencio, but is not limited thereto.

There are many causes of anticancer drug resistance, and among them, the tumor microenvironment has recently become a subject of interest. Tumors themselves have very complex heterogeneity and interact with a wide variety of cells around them. 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 organically connected to each other and are closely related to anticancer drug resistance (M.R. Junttila, Nature 501, 346, 2013). For example, important immune cells in the tumor microenvironment include regulatory T cells (Treg), myeloid-derived suppressor cells (MDSC), fibroblasts (cancer-associated fibroblasts (CAF)), and natural killers. There are cells (NK cells, Natural killer cells), etc.

In this regard, it is known that when STAT 3 is activated, it regulates the functions of various immune cells such as natural killer cells in the tumor microenvironment, thereby weakening the anticancer immune effect (H. Yu, Nature Reviews Immunology 7, 41, 2007).

The novel compound of the present invention or its pharmaceutically acceptable salt exhibits the effect of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by inhibiting STAT3 activity, thereby preventing the tumor microenvironment from responding to anticancer immunotherapy agents. It can be useful as a drug that overcomes the resistance of anticancer drugs and increases their activity by activating anticancer immunity, converting cancer (cold tumor) into cancer (hot tumor) that responds to immunotherapy drugs.

Another aspect of the present invention is a method for preventing or treating cancer, comprising administering to an entity other than a human a pharmaceutical composition containing the compound represented by Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient. Provides a method.

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

As used herein, the term "individual" may mean, but is not limited to, any animal such as a cow, horse, sheep, pig, goat, camel, antelope, dog, cat, or human that has developed or is likely to develop cancer. does not

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

The term "administration" of the present invention means introducing the composition of the present invention into an individual who is or is likely to exhibit symptoms of cancer and related diseases by any appropriate method, and the route of administration of the composition of the present invention is the target tissue. It can be administered through various routes, either oral or parenteral, as long as it can reach.

Specifically, the composition of the present invention is not particularly limited thereto, but may be administered by 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. It can be administered through.

Specifically, the dosage of the pharmaceutical composition of the present invention may be, for example, 0.1 to 500 mg/kg of 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, but may be administered once a day or divided into multiple doses. However, since it may increase or decrease depending on the route of administration, severity of disease, gender, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

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.

Another aspect of the present invention provides a food composition for preventing or alleviating cancer, comprising the compound represented by Formula 1 above, or a foodologically acceptable salt thereof, as an active ingredient.

The compound represented by Formula 1, active ingredients, cancer prevention and treatment are the same as described above.

The term “foodologically acceptable salt” of the present invention is as defined in “pharmaceutically acceptable salt” above.

The term “improvement” in the present invention means any action that reduces at least the degree of symptoms, for example, parameters related to the condition being treated by administration of the composition according to the present invention.

As used herein, “food” refers to meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, and alcoholic beverages. , vitamin complexes, health functional foods, and health foods, etc., and include all foods in the conventional sense.

Since the food composition of the present invention can be consumed on a daily basis, it can be expected to have a high cancer-improving effect, so it can be very useful for health promotion purposes.

Additionally, the food composition of the present invention can be used as a health functional food. The above “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Health Functional Food Act, and “functional” refers to food that is related to the structure and function of the human body. It means ingestion for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects. The food of the present invention can be manufactured by a method commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry. In addition, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in a variety of dosage forms, and unlike general drugs, it is made from natural products and has the advantage of not having side effects that may occur when taking the drug for a long period of time. It is also highly portable, so it is easy to use. The food of the invention can be consumed as a supplement to improve the effect of improving Parkinson's disease.

The above-mentioned health food refers to food that has a more active health maintenance or promotion effect compared to general food, and health supplement food refers to food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement are used interchangeably.

Specifically, the health functional food is a food manufactured by adding the extract of the present invention to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending it, and consuming it may cause certain health problems. It means bringing about an effect, but unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food.

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

In addition, the food composition may contain additional ingredients that are commonly used in food compositions to improve smell, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Additionally, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition contains preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxide) roxitoluene (BHT), etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin) , sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengtheners, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. Food May contain food additives. The additives may be selected depending on the type of food and used in an appropriate amount.

The food composition of the present invention may contain ordinary food additives, and its suitability as a "food additive" is determined in accordance with the general provisions of the Food Additives Code and General Test Methods approved by the Ministry of Food and Drug Safety, unless otherwise specified. The decision is made based on the specifications and standards for the item.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as reduced pigment, licorice extract, crystalline cellulose, high-lyin pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.

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

In addition, the food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing or improving cancer.

For example, the health functional food in the form of a tablet is prepared by granulating a mixture of the compound of Chemical Formula 1 or a foodologically acceptable salt thereof, excipients, binders, disintegrants, and other additives in a conventional manner, and then adding a lubricant. The mixture can be compression molded or the mixture can be compressed directly. In addition, the health functional food in the form of tablets may contain a flavoring agent, etc., if necessary, and may be peeled with a suitable peeling agent, if necessary.

Hard capsules, which are health functional foods in the form of capsules, are ordinary hard capsules filled with a mixture of powdered compounds of formula 1 or their food-acceptable salts and additives such as excipients, or granules thereof or peeled granules. Soft capsules can be manufactured by filling a mixture of the compound of Chemical Formula 1 or its food-acceptable salt and additives such as excipients into a capsule base such as gelatin. The soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary.

Health functional foods in the form of pills can be prepared by molding the compound of Chemical Formula 1 or a mixture of its foodologically acceptable salts, excipients, binders, disintegrants, etc. in an appropriate manner, and if necessary, coated with white sugar or other suitable coating agent. Alternatively, it may be coated with starch, talc, or a suitable substance.

Health functional foods in the form of granules can be manufactured into granules from the compound of Chemical Formula 1 or a mixture of its foodologically acceptable salts, excipients, binders, disintegrants, etc., using an appropriate method. Flavoring agents, flavoring agents, etc. may be added as needed. It may contain. Health functional foods in granular form use No. 12 (1680 μm), No. 14 (1410 μm), and No. 45 (350 μm) sieves. When performing the next particle size test, the entire amount passes through the No. 12 sieve and what remains on the No. 14 sieve is It may be 5.0% or less of the total amount, and what passes through the No. 45 sieve may be 15.0% or less of the total amount.

Definitions of terms such as excipients, binders, disintegrants, lubricants, coagulants, flavoring agents, etc. are described in literature known in the art and may include those with the same or similar functions (Korean Pharmacopoeia Commentary, Moonseongsa) , Korean Association of Colleges of Pharmacy, 5th revised edition, p33-48, 1989).

Another aspect of the present invention provides a feed composition for preventing or improving cancer, comprising the compound represented by Formula 1 above, or a foodologically acceptable salt thereof, as an active ingredient.

The compound represented by Formula 1, its food-acceptable salt, active ingredients, cancer prevention and improvement are the same as described above.

As used herein, the term "feed" refers to food consumed by animals, and may specifically mean a substance that supplies organic or inorganic nutrients necessary to maintain the life of an animal or to produce meat, milk, etc. . The feed may contain feed additives and can be manufactured in various forms known in the art.

The composition containing the compound of formula 1 or a foodologically acceptable salt thereof of the present invention can be used to prevent or treat cancer in non-human subjects such as livestock or companion animals, and can be used as a functional feed additive or for feed. It can be used as a composition.

The content of the compound of formula 1 or its foodologically acceptable salt in the feed composition according to the present invention can be appropriately adjusted depending on the type and age of the applied livestock, application form, desired effect, etc., for example, 1 to 99% by weight, preferably. Preferably, it can be used at 10 to 90% by weight, more preferably at 20 to 80% by weight, but is not limited thereto.

The type of feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include vegetable feed such as grains, roots and fruits, food processing by-products, algae, fibers, oils, starches, cucurbits or grain by-products; Examples include animal feed such as proteins, inorganic substances, fats and oils, minerals, fats and oils, single-cell proteins, zooplanktons, or food. These may be used alone or in combination of two or more types.

Alternatively, the compound of the present invention or a foodologically acceptable salt thereof may be used as a feed additive added to a feed composition. The above feed additives may be intended to improve the productivity or health of target animals, but are not limited thereto. The above feed additives may correspond to supplementary feed under the Feed Management Act.

The feed additive of the present application is a mixture of one or more ingredients selected from the group consisting of organic acids such as citric acid, humalic acid, adipic acid, and lactic acid, and natural antioxidants such as polyphenol, catechin, tocopherol, vitamin C, green tea extract, chitosan, and tannic acid. It can be used, and other common additives such as buffers and bacteriostatic agents can be added as needed. Additionally, it can be formulated into liquid, capsule, granule, or tablet as needed.

The above feed or feed additive may additionally contain substances that have various effects, such as supplementing nutrients and preventing weight loss, improving the digestibility of fiber in the feed, improving milk quality, preventing reproductive disorders and improving conception rates, and preventing high temperature stress in the summer. For example, various supplements such as amino acids, inorganic salts, vitamins, antioxidants, anti-fungal agents, microbial agents, etc., vegetable protein feeds such as ground or crushed wheat, barley, corn, animal protein feeds such as blood meal, meat meal, and fish meal, In addition to the main ingredients such as animal fat and vegetable fat, it can be used as a nutritional supplement, growth promoter, digestion and absorption enhancer, and disease prevention agent.

The feed and feed additives of the present application can be fed to a number of animals, including mammals and poultry. For example, to maintain or improve aesthetics, livestock such as cows and goats; It can be used on pets such as dogs and cats, but is not limited to this.

Another aspect of the present invention provides the use of a composition comprising the compound represented by Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient for the prevention or treatment of cancer.

The compound represented by Formula 1, its pharmaceutically acceptable salt, active ingredients, cancer, prevention and treatment are the same as described above.

The novel compound of the present invention or its pharmaceutically acceptable salt exhibits the effect of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by inhibiting STAT3 activity, and can therefore be usefully used in the prevention or treatment of cancer. there is.

Figure 1 shows ¹ H-NMR data of compound CG191 according to an embodiment of the present invention.
Figure 2 is a diagram showing that Stat 3 phosphorylation is inhibited in a prostate cancer cell line by the compound CG191 of the present invention in a concentration-dependent manner.
Figure 3 is a diagram showing that Stat 3 phosphorylation is inhibited in a time-dependent manner in cancer cell lines by the compound CG191 of the present invention.
Figure 4 is a diagram showing that Stat 3 phosphorylation is inhibited in breast cancer cell lines by the compound CG191 of the present invention in a concentration-dependent manner.
Figure 5 is a diagram showing that Stat 3 phosphorylation is inhibited in lung cancer cell lines by the compound CG191 of the present invention in a concentration-dependent manner.
Figure 6 is a diagram showing the results of measuring the efficacy of the compounds CG191, HCA, EA and HCA-EA mixture of the present invention to inhibit Stat 3 phosphorylation in cancer cell lines.
Figure 7 is a diagram showing the results of measuring the cancer cell growth inhibitory activity of the compound CG191 and HCA-EA mixture of the present invention in prostate cancer cell lines.
Figure 8 is a diagram showing the results of measuring the cancer cell growth inhibitory activity of the compound CG191 of the present invention in breast cancer cell lines.
Figure 9 is a diagram showing that the expression of Stat 3 target proteins is suppressed and cell death is induced in cancer cell lines by the compound CG191 of the present invention.
Figure 10 is a diagram showing that Stat 3 phosphorylation is inhibited in immune cells by the compound CG191 of the present invention.
Figure 11 is a diagram showing the results of measuring body weight changes in mice subcutaneously implanted with the compound CG191 of the present invention and the HCA-EA mixed drug for prostate cancer.
Figure 12 is a diagram showing the results of measuring the effect of the compound CG191 and HCA-EA mixture of the present invention on inhibiting the growth of cancer tissue in mice subcutaneously implanted with prostate cancer.
Figure 13 is a diagram showing the results of measuring the effect of the compound CG191 and HCA-EA mixture of the present invention on inhibiting the growth of cancer tissue in mice subcutaneously implanted with prostate cancer.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Synthesis Example 1: Synthesis of CG191

CG191, a compound of Formula 1, was synthesized using the method shown in Scheme 1 below.

[Scheme 1]

Specifically, 2-Hydroxycinnamaldehyde (400 mg) and ethacrynic acid (800 mg) were dissolved in DMF (N,N-dimethylformamide, 20 mL) and then N -Ethyl- N - (3-dimethylaminopropyl)carbodiimide hydrochloride (EDC, 700 mg) was added and stirred at room temperature for 16 hours. After adding water to terminate the reaction, extraction was performed three times using ethyl acetate and water. Afterwards, the organic layer was washed three times with water and then washed once more with salt water. This was dried using MgSO ₄ , filtered, and concentrated under reduced pressure. The concentrate was purified using column chromatography (MeOH: CHCl ₃ = 3: 97) to obtain compound 1 [(E)-2(3-oxoprop-1-en-1-yl)phenyl 2-[2,3- dichloro-4-(2-methylenebutanoyl)phenoxy]acetate, 650 mg, 57.6%, white powder] was obtained.

The mp of compound 1 is 106°C, and ^{the 1H} , ¹³ C-NMR analysis results are as follows.

¹ HNMR (400MHz, CDCl ₃ )δ9.67 (d, J = 7.6 Hz, 1H), 7.68 (dd, J = 1.6, 7.6 Hz, 1H), 7.55 (d, J = 16 Hz, 1H), 7.49 ( dt, J = 1.6, 7.6 Hz, 1H), 7.35 (t, J = 7.2 Hz, 1H), 7.23 (d, J = 7.2 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 6.93 ( d, J = 6.9 Hz, 1H), 6.73 (dd, J = 7.6, 16 Hz, 1H), 5.95 (s, 11H), 5.59 (s, 1H), 5.07 (s, 2H), 2.47 (q, J = 7.6 Hz, 2H), 1.51 (t, J = 7.2 Hz, 3H);

¹³ CNMR (100MHz, CDCl ₃ )δ195.58, 193.21, 165.86, 155.03, 150.12, 148.36, 144.76, 134.56, 132.17, 131.89, 130.68, 128.78, 128.19, 1 27.08, 126.81, 126.56, 123.67, 122.76, 110.96, 66.38, 23.36, 12.35.

The compound synthesized as above was named CG191, and its ¹ H-NMR data is shown in Figure 1.

Experimental Example 1: Cell Culture

All cell lines used to verify the in vitro activity of CG191 prepared in Synthesis Example 1 were purchased from ATCC (Manassas, VA, USA). Human prostate cancer cell line DU-145, breast cancer cell MDA-MB 468, and lung cancer cell HCC827 were used in RPMI-1640 (Gibco, Gaithersburg, MD, USA) medium containing 10% FBS (Gibco) and 100u/ml penicillin. Natural killer cell NKL cell line was used in RPMI-1640 (ATCC, 30-2001) medium with 10% FBS (Gibco) and 50 ng/ml hIL-2 (Peprotech, East Windsor, NJ, USA). All cell lines used in the experiment were maintained in a 5% CO ₂ incubator.

Experimental Example 2: Western blot analysis

Prostate cancer cell line DU-145, breast cancer cell MDA-MB 468, and lung cancer cell HCC827 (4 x 10 ⁵ cells) were each plated in a 6 well plate. After 24 hours, cells were treated with 0.1% DMSO, HCA, EA, and CG-191 at different concentrations and times, washed with PBS, and recovered using RIPA buffer. 20 g of protein quantified using the Bio-Rad protein assay kit (Bio-Rad, Hercules, California, USA) was subjected to 8% SDS-PAGE electrophoresis and then analyzed using a PVDF membrane (EMD Millipore, Billerica, MA, USA). Transfer was carried out. After blocking the membrane with TBST containing 5% skim milk, the experiment was 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), and Survivin (Cell Signaling). , Bcl-xL (Santa Cruz), PARP (Cell Signaling) GAPDH (Santa Cruz), and the secondary antibodies are HRP-conjugated goat anti-rabbit, anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). used. The antibodies used were each washed three times with TBST, and bands were detected using Luminata Forte Western HRP substrate (EMD Millipore) reagent and LAS 4000 mini (GE Healthcare Life Sciences) device. Subsequently, correction using GAPDH was performed using the MultiGauge program (Fuji Photo Film, Tokyo, Japan).

Example 1: Confirmation of Stat 3 phosphorylation inhibition effect in cancer cell lines

The inhibitory effect of CG191 prepared in Synthesis Example 1 on Stat 3 phosphorylation in cancer cell lines was confirmed by Western blot.

As a result, as shown in Figures 2 and 3, it was confirmed that treatment with the compound CG191 of the present invention inhibited Stat 3 activity (p-STAT3) in a concentration-dependent manner in prostate cancer cells DU-145, especially when treated at 10 μM. It was confirmed that STAT 3 activity (p-STAT3) was inhibited by more than 50% (Figure 2). In addition, it was confirmed that treatment with the compound CG191 of the present invention inhibited STAT 3 activity (p-STAT3) in a time-dependent manner in prostate cancer cells DU-145 (FIG. 3).

In addition, as shown in Figures 4 and 5, it was confirmed that treatment with the compound CG191 of the present invention inhibits Stat 3 activity (p-STAT3) in a concentration-dependent manner in breast cancer cells and lung cancer cells, respectively. In particular, when treated with 30 μM, STAT 3 activity was confirmed. 3 It was confirmed that activity (p-STAT3) was inhibited by more than 90%.

In summary, it was confirmed that treatment with the compound CG191 of the present invention inhibits STAT 3 activity (p-STAT3) in a concentration-dependent manner in all representative carcinoma cells, such as prostate, breast, and lung cancer cells.

Next, 2-hydroxycinnamaldehyde (HCA), ethacrynic acid (EA), and HCA-EA mixed drug (simple mixture of HCA and EA) and the above synthesis example in prostate cancer cell line, which is a representative carcinoma. The inhibitory effect of CG191 prepared in step 1 on Stat 3 phosphorylation was confirmed by Western blotting and compared.

As a result, as shown in Figure 6, it was confirmed that treatment with the compound CG191 of the present invention exhibited a significant Stat 3 phosphorylation inhibitory effect compared to HCA, EA, and HCA-EA mixed drugs.

Example 2: Confirmation of cancer cell growth inhibition effect due to inhibition of Stat 3 phosphorylation in cancer cell lines

In Example 1, it was confirmed that treatment with the compound CG191 of the present invention inhibits Stat 3 activity (p-STAT3) in cancer cells. In order to confirm the effect of Stat 3 activity inhibition on cancer cell growth inhibition, Cell proliferation assay was performed using prostate cancer cell lines and breast cancer cells.

Specifically, prostate cancer cell line DU-145 and breast cancer cell MDA-MB 468 (0.8 x 10 ⁴ cells) were each plated in a 96 well plate. After 24 hours, DMSO (control group), HCA-EA mixed drug, and CG-191 were treated at each concentration for 24 or 48 hours, and then 10 ul of WST-1 (Dojindo Laboratories, Kumamoto, Japan) was treated in each well. After treatment for about 1 hour, cell viability was calculated using the absorbance measured at 450 nm with a microplate reader (Multiskan GO system, Thermo Fisher Scientific, Rockford, IL, USA).

As a result, as shown in Figure 7, it was confirmed that the compound CG191 of the present invention inhibits the growth of prostate cancer cells in a concentration-dependent manner. Additionally, as shown in Figure 8, it was confirmed that the compound CG191 of the present invention inhibits the growth of breast cancer cells in a concentration-dependent manner.

Accordingly, it was confirmed that the compound CG191 of the present invention inhibits the growth of cancer cells in a concentration-dependent manner. In addition, it was confirmed that it exhibited a significant cancer cell growth inhibition effect compared to the HCA-EA mixed drug (Figure 7).

Accordingly, it was confirmed that the compound CG191 of the present invention inhibits the growth of cancer cells by inhibiting the activity of Stat 3 in cancer cells.

Example 3: Confirmation of inhibition of expression and cell death effect of Stat 3 target proteins in cancer cell lines

In Example 1, it was confirmed that treatment with the compound CG191 of the present invention inhibits Stat 3 activity (p-STAT3) in cancer cells. The effect of inhibiting the expression of Stat 3 target proteins and the cell death effect due to inhibition of Stat 3 activity were tested using Western methods. Confirmed by blot.

As a result, as shown in Figure 9, it was confirmed that the compound CG191 of the present invention significantly inhibits the expression of the target proteins of stat 3, Cyclin D1, Cyclin A, Mcl-1, Survivin, and Bcl-xL. In addition, it was confirmed that CG191 exerts an anti-cancer effect through cell death by confirming that it promotes the degradation of PARP protein, a cancer cell death biomarker.

Example 4: Confirmation of Stat 3 phosphorylation inhibition effect in immune cells

The inhibitory effect of Stat 3 phosphorylation of CG191 prepared in Synthesis Example 1 in natural killer cells (NKL), which are immune cells, was confirmed by Western blot.

Specifically, NKL ( ⁴ After treatment with 10 ng/ml of IL-6, one of the kines, for 30 minutes, treatment with 20 uM of PBS or CG191 for 30 minutes, the cells were recovered using a centrifuge. The recovered cells were washed once with PBS and p-STAT3 expression was measured by western blot.

As a result, as shown in Figure 10, it was confirmed that treatment with the compound CG191 of the present invention inhibited Stat 3 activity (p-STAT3) in immune cells by more than 30%, and Stat 3 activated with IL-6, an activating cytokine. It was confirmed that the activity of 3 was also inhibited by treatment with the compound CG191 of the present invention.

Example 6: Confirmation of growth inhibition effect of cancer tissue in nude mice

We sought to evaluate the anticancer efficacy of the compound CG191 of the present invention by intraperitoneal administration through a nude mouse subcutaneous transplant model of human-derived prostate cancer (DU-145).

Specifically, the animals used were Athymic-NCr NCr-nu specific pathogen-free (SPF) 5-week-old nude mice supplied by Coretech (Pyeongtaek-si, Gyeonggi-do). The cancer cell concentration was adjusted to 3 × 10 ⁷ cells/ml, and 0.3 mL of cell culture per mouse was injected subcutaneously into the axillary area between the right shoulder and chest wall. Next, the HCA-EA mixed drug or CG191 was prepared at the corresponding concentration by adding 10% DMAC + 10% Tween80 + 20% HPbCD 80% in stages, and was administered intraperitoneally 0.2 ml per 20 g of mice a total of 19 times over 25 days. . 25 days after starting drug administration, the DU-145 tumor was excised and its weight was measured.

As a result, as shown in Figure 11, it was confirmed that CG191 did not induce body weight change.

In addition, as shown in Figures 12 and 13, as of the final day (day 25), tumor growth inhibition of 60% (p<0.001) was confirmed in the CG191 administration group compared to the solvent control group (Vehicle). In addition, it was confirmed that it exhibited a significant anti-tumor agent effect compared to the HCA-EA mixed drug (40% inhibition, p<0.001) (FIG. 12).

Additionally, as of the final day (day 25), a 67.2% (p<0.001) reduction in tumor weight was confirmed in the CG191 administration group compared to the solvent control group (Vehicle). In addition, it was confirmed that it showed a significant tumor weight reduction effect compared to the HCA-EA mixed drug (46.8% reduction, p<0.001) (FIG. 13).

Accordingly, it was confirmed that the compound CG191 of the present invention effectively inhibits the growth of cancer tissue in animal models.

In summary, it was confirmed that the compound CG191 of the present invention inhibits the activity of Stat 3 in cancer cells, suppresses the expression of target molecules of Stat 3 and induces cell death. In addition, it was confirmed that the anti-cancer effect can be increased by controlling the activity of immune cells by confirming the inhibition of the activity of Stat 3 in natural killer cells, which are immune cells. In addition, it was confirmed that it effectively inhibits the growth of cancer tissue in animal models.

Accordingly, the novel compound of the present invention or its pharmaceutically acceptable salt exhibits the effect of inhibiting the growth of cancer cells and increasing the activity of immune cells in the tumor microenvironment by inhibiting STAT3 activity, and is thus useful for the prevention or treatment of cancer. It was confirmed that it can be used.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Claims (11)

A compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof:
[Formula 1]
.
A pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof, as an active ingredient:
[Formula 1]
.
The composition of claim 2, wherein the composition inhibits the activity of signal transducers and activators of transcription 3 (STAT3).
The method of claim 2, wherein the cancer is colon cancer, pancreatic cancer, stomach 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 lymphoma, and Hodgkin lymphoma. , blood cancer, bladder cancer, kidney cancer, ovarian cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer. , selected from the group consisting of endocrine 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, brainstem glioma, and pituitary adenoma. A composition that is any one of the following.
The composition of claim 2, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent.
The composition of claim 2, wherein the composition further comprises an anticancer agent.
The method of claim 6, wherein the anti-cancer agent consists of DNA alkylating agents, anti-cancer antibiotics, plant alkaloids, targeted anti-cancer agents, and cancer immunotherapy. A composition comprising at least one selected from the group.
The method of claim 6, wherein the anticancer agent is mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, car Carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (dactinomycin: actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin, C Bleomycin; Vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, iridotecan, glivec ), tasigna, Herceptin, iressa, nexsavar, yervoy, opdovo, keytruda, Tecentriq, Imfinzi ) and a composition that is at least one selected from the group consisting of babencio.
A method for preventing or treating cancer, comprising administering to an entity other than a human a pharmaceutical composition containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]
.
The method of claim 9, wherein the pharmaceutical composition inhibits the activity of signal transducers and activators of transcription 3 (STAT3).
A food composition for preventing or ameliorating cancer, comprising a compound represented by the following formula (1), or a foodologically acceptable salt thereof, as an active ingredient:
[Formula 1]
.