KR102544591B1 - Novel compound and their uses - Google Patents

Abstract

The present invention relates to a novel mifepristone derivative compound and various uses thereof.
In order to develop an effective cancer treatment, it is necessary to simultaneously inhibit the growth and metastasis of cancer cells. In general, cancer growth refers to the characteristics of infinitely proliferating cancer cells, and cancer metastasis refers to the characteristics of cancer cells being separated from a primary site and moving to another place. Therefore, cancer growth and metastasis require approaches from different mechanisms.
Since the compound provided in the present invention can simultaneously inhibit the proliferation and metastasis of cancer, it is expected to be actively used in the medical and health fields.

Description

Novel compounds and their uses {Novel compounds and their uses}

The present invention relates to a novel mifepristone derivative compound and various uses thereof.

Cancer is one of the most common causes of death worldwide. Approximately 10 million new cases occur each year, accounting for approximately 12% of all deaths, making it the third leading cause of death.

In order to develop an effective cancer treatment, it is necessary to simultaneously inhibit the growth and metastasis of cancer cells. In general, cancer growth refers to the characteristic of infinitely proliferating cancer cells. Normal cells stop dividing when they finish dividing as much as they were programmed into their genes, or when there is not enough room for new cells to occupy. However, cancer cells fail to regulate the function of controlling division, and when space to expand themselves is insufficient, they secrete a proteolytic enzyme (matrix metalloprotease) that decomposes the cell matrix to remove the surrounding space and create space for themselves. expand On the other hand, cancer metastasis refers to a characteristic in which cancer cells are separated from a primary site and move to another place. To do this, ① cancer cells must be shed from the primary site (cancer tissue), ② invade and pass through the tissue walls constituting blood vessels or lymphatics, and ③ adhere to and fuse with the matrix of the migrated tissue to settle in a new place. However, an approach from a mechanism different from cancer growth is needed.

Therefore, the present invention has been made to solve the above problems, and relates to a novel compound capable of simultaneously inhibiting cancer proliferation and metastasis, and pharmaceutical use thereof.

One object of the present invention is to provide novel compounds capable of preventing, improving or treating cancer.

Another object of the present invention is to provide a composition capable of preventing, improving or treating cancer, including the above compound.

Another object of the present invention is to provide a method for preventing, improving or treating cancer using the above compounds.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, numerous specific details are set forth, such as specific forms, compositions and processes, etc., in order to provide a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, the appearances of "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, particular features, forms, compositions, or properties may be combined in one or more embodiments in any suitable way.

Unless otherwise defined in the specification, all scientific and technical terms used herein have the same meaning as commonly understood by a person skilled in the art to which the present invention belongs.

In the present invention, "cancer" is characterized by uncontrolled cell growth, and due to such abnormal cell growth, a cell mass called a tumor is formed, penetrates into surrounding tissues, and in severe cases, metastasizes to other organs of the body. means that Scientifically, it is also called neoplasia. Cancer is an intractable chronic disease that in many cases cannot be fundamentally cured even if treated with surgery, radiation, and chemotherapy, causing pain to patients and ultimately leading to death. There are many causes of cancer, but internal factors and external factors. Although it has not been precisely clarified how normal cells are transformed into cancer cells through any mechanism, it is known that a significant number of cancers are affected by external factors such as environmental factors. Internal factors include genetic factors and immunological factors, and external factors include chemicals, radiation, and viruses. Genes involved in the development of cancer include oncogenes and tumor suppressor genes, and cancer occurs when the balance between these genes is disrupted by the internal or external factors described above. The cancer is preferably breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, larynx cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, etc., and cancer progression, such as tumor differentiation and / or proliferation, is limited to the type of cancer dependent on cancer cells and / or cancer stem cells described in the present invention It doesn't work. At this time, the cancer stem cell (CSC) refers to cancer cells in a comprehensive sense that have self-renewal ability or differentiation ability, which is a unique ability of stem cells, and cancer stem cells in cancer tissues account for 0.1 to 5%. It is presumed to exist between The cancer stem cells are under normal tumor growth conditions (the "normal tumor growth conditions" refer to a state in which there is no cell stress due to sufficient nutrients (glucose) necessary for cell growth and sufficient growth conditions in the tumor microenvironment). Unlike normal cancer cells, it has a characteristic of slow proliferation. In addition, it may have resistance to anticancer drugs by maintaining a dormant state, and unlike normal tumor cells, the expression of transcriptional regulators such as PGC-1α is controlled, so that the function of major metabolic regulators is higher than that of normal cancer cells. There are different characteristics. It comprehensively refers to cells with invasion and/or metastatic ability by acquiring resistance to cell death (apoptosis) in a nutrient deprivation state through the regulation of these different metabolisms and the cell signaling system associated therewith. However, it is not limited thereto as long as it is a cell capable of differentiating into a general cancer cell.

In the present invention, "virtual screening" is one of the many procedures that go through during the new drug development process, and more specifically, the process of discovering effective substances and lead substances through screening of selected compounds from basic research and confirming candidate substances. say This can reduce the time and cost required for new drug development, and after a target disease is selected, it is possible to discover an effective substance for a new drug targeting it. It is also possible to selectively find materials that are The virtual screening technology can be used not only for library production but also for active screening for actual libraries and specific action points. However, if it is a general and universal screening technique, it is not limited thereto.

In the present invention, the compound of formula A below was derived as an anti-cancer candidate through virtual screening as described above and a derivative thereof was synthesized.

[Formula A]

The derivative of Formula A may be a compound represented by Formula B below.

[Formula B]

However, in a preferred embodiment, the derivative of Chemical Formula A may be a novel compound synthesized in the present invention and represented by Chemical Formulas 1 to 6 below.

[Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

According to Scheme 1 described in the present invention, Formula 1 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methyl-enbis(oxy)] (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis synthesizing (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxyl-16beta-methyl-17,20:20,21-bis[methylene Bis(oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy )]) synthesizing; and (c) pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxyl,17-alpha,21-dihydroxy-10; 13,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxy,17-alpha,21-dihydroxy-10,13 , synthesizing 16-beta-trimethyl); to be prepared by a manufacturing method comprising, pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta (4'-chloro)-benzoxyl-16-beta-methyl-17-hydroxy (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'- chloro)-benzoxy-16-beta-methyl-17-hydroxy).

According to Scheme 2 described in the present invention, Formula 2 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methyl-enbis(oxy)] (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis synthesizing (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-benzoxyl-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] (Pregna synthesizing -1,4-dien-3-one,9-fluoro-11beta-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)]); and (c) pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxyl, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl ( Synthesis of Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl) prepared by the method, pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta benzoxyl,17-hydroxy-10,13,16-beta- It can be named trimethyl (Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta benzoxy, 17-hydroxy-10,13, 16-beta-trimethyl).

According to Scheme 3 described in the present invention, Formula 3 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methyl-enbis(oxy)] (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis synthesizing (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxyl-16beta-methyl-17,20:20,21-bis[ Methylenebis(oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis( synthesizing oxy)]); and (c) pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-isopropyl)-benzoxyl, 17-alpha, 21-dihydroxy-10 ,13,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-isopropyl)-benzoxy, 17-alpha, 21-dihydroxy-10, Synthesizing 13,16-beta-trimethyl); Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11- Beta-(4'-isopropyl)-benzoxyl,17-hydroxy-10,13,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro- 11-beta-(4'-isoproyl)-benzoxy,17-hydroxy-10,13,16-beta-trimethyl).

According to Scheme 4 described in the present invention, Formula 4 is (a) preg-4-ene-3,20-dione, 10,13-dimethyl-17,21-[(1-methoxypropylidene)bis(oxy )] (Pregn-4-ene-3,20-dione,10,13-dimethyl-17,21-[(1-methoxypropylidene)bis(oxy)]); and (b) Pregn-4-ene-3,20-dione-10,13-dimethyl-21-chloro-17-hydroxy-17-propionate (Pregn-4-ene-3,20-dione- Synthesizing 10,13-dimethyl-21-chloro-17-hydroxy-17-propionate); Preg-1,4-diene-3,20-dione-10 ,13-dimethyl-21-chloro-17-hydroxy-17-propionate (Pregn-1,4-diene-3,20-dionee-10,13-dimethyl-21-chloro-17-hydroxy-17- propionate).

Formula 5 is synthesized from mifepristone according to Scheme 5 described in the present invention, and 11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl)-delta 4,9-estradien-3-one-17beta-hydroxy-(2'-chloro)acetate (11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl )-delta4,9-estradiene-3-one-17beta-hydroxy-(2'-chloro)acetate).

According to Scheme 6 described in the present invention, Formula 6 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methyl-enbis(oxy)] (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis synthesizing (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxyl-16beta-methyl-17,20:20,21-bis[methylene Bis(oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy )]) synthesizing; (c) Pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxyl,17-alpha,21-dihydroxy-10,13 ,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxy,17-alpha,21-dihydroxy-10,13, synthesizing 16-beta-trimethyl); and (d) pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro)-benzoxy-16-beta-methyl-17-hydroxy Synthesis of hydroxyl (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro)-benzoxy-16-beta-methyl-17-hydroxy) ; Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta (4'-chloro-benzoxyl) -16 -beta-methyl-17-hydroxypropionate (Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta (4'-chloro-benzoxy) -16-beta -methyl-17-hydroxy propionate).

The present invention provides the novel compound described above, and a method for preparing (synthetic method) thereof, and also provides a pharmaceutically acceptable salt of the compound. Such pharmaceutically acceptable salts are salts generally regarded by those skilled in the art as being suitable for medical applications (eg because such salts are not detrimental to the subject being treated with the salts), or within the respective treatment It is a salt that causes acceptable side effects in Generally, the pharmaceutically acceptable salt is a salt that is considered acceptable by regulatory authorities such as the US Food and Drug Administration (FDA), the European Medicines Agency (EMA), or the Pharmaceuticals and Medical Devices Agency (PMDA) of the Japanese Ministry of Health, Labor and Welfare. . However, the present invention in principle relates to, for example, an intermediate in the preparation of a compound according to the invention or a physiologically functional derivative thereof, or a pharmaceutically acceptable salt of a compound according to the invention or a physiologically functional derivative thereof. As intermediates in the preparation of derivatives, salts of the compounds according to the invention which are not pharmaceutically acceptable per se are also included. The salts include water-insoluble salts and, in particular, water-soluble salts.

In each case, the person skilled in the art can determine whether a particular compound according to the present invention or a physiologically functional derivative thereof can form a salt, i.e., whether said compound according to the present invention or a physiologically functional derivative thereof For example, it can be easily determined whether or not it has a group capable of carrying an electric charge, such as an amino group or a carboxylic acid group.

Exemplary salts of the compounds of the present invention are acid addition salts or salts with bases, particularly pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases commonly used in pharmaceutics, which are water insoluble or particularly water soluble acid addition salts. It is salt. Depending on the substituents of the compounds of this invention, salts with bases may also be suitable. Acid addition salts are formed, for example, by combining a solution of a compound of the present invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid, or phosphoric acid. It can be formed by mixing. Likewise, pharmaceutically acceptable base addition salts include alkali metal salts (eg sodium or potassium salts); alkaline earth metal salts (eg, calcium or magnesium salts); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium and amines formed using counter anions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl sulfonates and aryl sulfonates). cations) may be included. Illustrative examples of pharmaceutically acceptable salts include acetates, adipates, alginates, arginates, ascorbates, aspartates, benzenesulfonates, benzoates, bicarbonates, bisulfates, bitartrates, borates, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, digluconate, dihydrochloride, dodecylsulfate, edetate, edisylate, ethanesulfonate, Formate, fumarate, galactate, galacturonate, gluconate, glutamate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrobromide, hydrochloride, hydroiodide , 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide, isobutyrate, isothionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, methane Sulfonate (mesylate), methyl sulfate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate /diphosphate, phthalate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, suberate, succinate, tannate, tartrate, tosylate, undecanoate, valerate and the like, but are not limited thereto.

Salts which are not pharmaceutically acceptable and which can be obtained as process products, for example during the preparation of the compounds according to the invention on an industrial scale, are also included in the present invention and, if desired, by methods known to the person skilled in the art. It can be converted into a pharmaceutically acceptable salt.

Meanwhile, since the compounds according to the present invention may have an asymmetric carbon center, they may exist as R or S isomers or racemic compounds, and all optical isomers and mixtures thereof may be included in the scope of the present invention.

In addition, the compounds of the present invention, as well as their salts, when isolated, for example in crystalline form, may contain varying amounts of solvent. Thus, solvates, especially hydrates, of the compounds of the present invention as well as solvates, particularly hydrates, of salts of the compounds of the present invention may be included within the scope of the present invention. More particularly, the present invention may include hydrates of the compounds, salts and/or physiologically functional derivatives according to the present invention comprising 1, 2 or 1/2 water molecules relative to the stoichiometry. .

Hereinafter, the compound of the present invention means the compound described above.

The compound of the present invention, a compound selected from pharmaceutically acceptable salts, crystalline forms, co-crystals, optical isomers, derivatives, hydrates, and solvates thereof can be used for the prevention, improvement, or treatment of a desired disease.

At this time, the target disease is skeletal disease, joint disease, vascular disease, blood disease, skin disease, kidney disease, cardiovascular disease, cranial nerve disease, ophthalmological disease, digestive disease, urinary disease, gynecological disease, respiratory disease, infectious disease, allergy disease, rheumatic disease, autoimmune disease, disease due to transplantation or transplant side effects, or proliferative disease (tumor).

The present invention provides a pharmaceutical composition comprising, as an active ingredient, a compound selected from the compound of the present invention, a pharmaceutically acceptable salt, crystalline form, co-crystal, optical isomer, derivative, hydrate, and solvate thereof.

In the present invention, the pharmaceutical composition can prevent, improve or treat cancer. Prevention, improvement, or treatment herein comprehensively includes activities that inhibit the generation or growth of cancer, or improve symptoms of cancer invasion, metastasis, or recurrence.

In the present invention, cancer as a target for improvement or treatment with a pharmaceutical composition containing the compound of the present invention as an active ingredient is liver cancer, bile duct cancer, gallbladder cancer, esophageal cancer, stomach cancer, ovarian cancer, breast cancer, uterine cancer, colon cancer, rectal cancer, Cervical cancer, prostate cancer, skin cancer, pancreatic cancer, leukemia, lymphoma, Hodgkin's disease, lung cancer, bronchial cancer, multiple myeloma, leukemia, lymphoma, squamous cell cancer, kidney cancer, urethral cancer, bladder cancer, head and neck cancer, brain cancer and central nervous system cancer It may be at least one selected from the group consisting of, preferably lung cancer, and more preferably non-small cell lung cancer.

In the present invention, the pharmaceutical composition may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be intended for humans.

The pharmaceutical composition of the present invention is not limited to these, but is formulated according to conventional methods into oral formulations such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories and sterile injection solutions. can The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavors, etc. for oral administration, and buffers, preservatives, and painless agents for injections. A topical, solubilizing agent, isotonic agent, stabilizer, etc. may be mixed and used, and in the case of topical administration, a base, excipient, lubricant, preservative, etc. may be used. Formulations of the pharmaceutical composition of the present invention may be variously prepared by mixing with the pharmaceutically acceptable carrier as described above. For example, for oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in unit dosage ampoules or multiple dosage forms. there is. In addition, it may be formulated into solutions, suspensions, tablets, capsules, sustained-release preparations, and the like.

On the other hand, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil and the like can be used. In addition, fillers, anti-coagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, and the like may be further included.

The route of administration of the pharmaceutical composition according to the present invention is not limited to these, but is oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical , sublingual or rectal. Oral or parenteral administration is preferred. The parenteral administration includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrabursal, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

The pharmaceutical composition of the present invention depends on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, route of administration, excretion rate, drug combination and severity of the specific disease to be prevented or treated. It can vary widely, and the dosage of the pharmaceutical composition varies depending on the patient's condition, body weight, disease severity, drug form, administration route and period, but can be appropriately selected by those skilled in the art, and is 0.0001 to 50 mg/day. kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated into a pill, dragee, capsule, liquid, gel, syrup, slurry, or suspension.

The pharmaceutical composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

In addition, the present invention provides a compound selected from the compound of the present invention, a pharmaceutically acceptable salt, crystalline form, co-crystal, optical isomer, derivative, hydrate, and solvate thereof to a subject (eg, human) in need of administration. It provides a method for preventing, improving or treating cancer, comprising administering a pharmaceutical composition comprising ingredients in a pharmaceutically effective amount. In this case, the above "administration" means providing a given compound of the present invention to a subject by any suitable method.

Specific details regarding the cancer, pharmaceutical composition, and route of administration in the method for preventing, improving, or treating cancer are described in the above “Compound of the present invention, pharmaceutically acceptable salt thereof, crystalline form, co-crystal, optical isomer, derivative , A hydrate, and a solvate as an active ingredient,” which overlaps with what is described in, and is omitted in order to prevent excessive complexity of the specification.

In the present invention, the "subject" in need of the administration may include both mammals and non-mammals. Here, examples of such mammals include humans, non-human primates such as chimpanzees, and other apes and monkey species; livestock animals such as cattle, horses, sheep, goats, pigs; domesticated animals such as rabbits, dogs and cats; Experimental animals, such as rodents, such as rats, mice, and guinea pigs, may be included, but are not limited thereto. In addition, examples of the non-mammals in the present invention may include birds and fish, but are not limited thereto.

In the present invention, the "pharmaceutically effective amount" refers to a sufficient amount of an agent to provide a desired biological result. The result is the reduction and/or alleviation of the signs, symptoms or causes of a disease, or any biological outcome. There may be other desired variations. For example, an "effective amount" for therapeutic use is the amount of a compound disclosed herein that is required to provide a clinically significant reduction in disease. In any individual case, an appropriate "effective amount" " Amounts can be determined by those skilled in the art using routine experimentation. Accordingly, the expression "effective amount" generally refers to an amount in which an active substance has a therapeutic effect.

The present invention also relates to a food composition for preventing or improving cancer comprising a compound selected from the compound of the present invention, a food acceptable salt, crystalline form, co-crystal, optical isomer, derivative, hydrate, and solvate thereof. to provide.

The food composition herein is used in various ways for the purpose of the present invention, that is, the prevention or improvement of cancer, and the food composition containing the compound of the present invention can be used in various foods, such as beverages, gum, tea, It can be manufactured in the form of vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, and bread. When the composition of the present invention is included in a food composition, the amount may be added in an amount of 0.1 to 100% of the total weight. Here, when the food composition is prepared in the form of a beverage, there is no particular limitation except that the food composition is contained in the indicated ratio, and various flavors or natural carbohydrates may be included as additional ingredients as in conventional beverages. In other words, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, and common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. can do. Examples of the flavoring agent include natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). Others of the present invention The food composition of is various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, Stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. These components may be used independently or in combination The ratio of these additives is usually per 100 parts by weight of the composition of the present invention It is generally selected from the range of 0.1 to 100 parts by weight, but is not limited thereto.

Hereinafter, the present invention will be described in detail based on examples.

Since the compound provided in the present invention can simultaneously inhibit the proliferation and metastasis of cancer, it is expected to be actively used in the medical and health fields.

Figure 1 shows the cancer cell survival inhibitory effect of the novel compound synthesized in the present invention according to one embodiment of the present invention.
Figure 2 shows the cancer cell tumor formation inhibitory effect of the novel compound synthesized in the present invention according to one embodiment of the present invention.
Figure 3 shows the cancer cell invasion inhibitory effect of the novel compound synthesized in the present invention according to one embodiment of the present invention.

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

Example 1: Preparation of novel anti-cancer compounds

The inventors of the present invention conducted virtual screening of various candidate substances in order to discover cancer cell and/or cancer stem cell treatment substances having anticancer activity, and as a result, derived the compound of formula A as a candidate substance for anticancer and synthesized a derivative thereof.

[Formula A]

In the following manufacturing example, TLC Silica Gel 60 GF254 from a Chinese manufacturer was used as the TLC plate, and the HPLC test method used a 4.6mm X 15cm column and C18 packing (pore size 5um). Refer to USP test conditions for clobetasol propionate did

Preparation Example 1 (S001)

Synthesis was performed according to Scheme 1 below.

[Scheme 1]

1. Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enbis(oxy)]( Synthesis of Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis(oxy)]).

10 g of betamethasone (Pregna-1,4-diene-3,20-dione, 9-fluoro-11beta,17alpha,21-trihydroxy-16beta-methyl) was dissolved in 100ml of dichloromethane. 200 ml of 7 mol/L hydrochloric acid and 20 g of paraformaldehyde were added, stirred for 24 hours, and the reaction was confirmed by TLC (eluent n-Hex:EA = 1:1 Rf = 0.4). When the reaction was complete, the organic layer was evaporated. Then recrystallized from methanol to obtain 6.7 g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxyl-16beta-methyl-17,20:20,21-bis[methylenebis (oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy) ]) Synthesis of.

6.7 g of the product of 1 (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methylenebis(oxy)]) was dissolved in tetrahydrofuran. dissolved in It was cooled at 0° C. and 1.2 g of sodium hydrogen (60%) was added. After stirring for 30 minutes, 2 g of benzyl bromide was dropped into the reaction solution and stirred for 24 hours. The reaction was quenched with water and purified by silica gel column chromatography (eluent n-Hex:EA = 9:1 to 5:1) to give 3.1 g of product.

3. Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-chloro)-benzoxyl, 17-alpha, 21-dihydroxy-10,13, 16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxy,17-alpha,21-dihydroxy-10,13,16 -beta-trimethyl) synthesis.

The product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) was dissolved in 20ml tetrahydrofuran, cooled at 0°C, and 20ml hydrofluoric acid was added and stirred for 14 hours. The organic layer was extracted with ethyl acetate, and the organic solvent was evaporated to obtain 2.6 g of product.

4. Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta (4'-chloro) -benzoxyl-16-beta-methyl-17-hydroxy ( Synthesis of Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro)-benzoxy-16-beta-methyl-17-hydroxy).

The product of 3 (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-chloro)-benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta -trimethyl) 2.6 g was dissolved in 5 ml pididine, cooled at 0 ° C, and then 700 mg of methanesulfonyl chloride was added and the reaction was completed by TLC (eluent n-Hex:EA = 2: 1 Rf = 0.35). Confirmed. After adding ethyl acetate to the residue and washing with water, the organic solvent was evaporated to give the crude product. It was dissolved directly in butanone, 2 g of anhydrous lithium chloride was added, and stirred for 24 hours. The organic layer was evaporated and purified by silica gel column chromatography (eluent n-Hex:EA = 5:1) to give 1.9 g of product. This is hereinafter referred to as the S001 compound or the compound of Formula 1.

[Formula 1]

The results of NMR analysis of the S001 compound (Formula 1) are as follows:

¹ H NMR (500 MHz, CDCl ₃ ) δ 7.343 (d, 2H, J=10.5 Hz), 7.265 (d, 2H, J=10.5 Hz), 6.82 (d, 1H, J=12.5 Hz), 6.297 (dd , 1H, J1=12.5Hz, J2=3Hz), 6.094 (s,1H),4.717(d,1H, J=13.5Hz), 4.487 (AB, 2H, J1=54.0Hz, J2=20.5Hz), 4.274 (d,1H, J=13.5Hz),4.03 (dm, 1H, J=11.0Hz), 1.389(s,3H),1.171(s,3H), 1.148 (d, 3H, J=7.5Hz),

LC-MS (ESI) m/z: 535.0 [M+H] ⁺

HPLC purity: 98.29%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=50:50

Production Example 2 (S002)

Synthesis was performed according to Scheme 2 below.

[Scheme 2]

1. Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enbis(oxy)]( Synthesis of Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis(oxy)]).

10 g of betamethasone (Pregna-1,4-diene-3,20-dione, 9-fluoro-11beta,17alpha,21-trihydroxy-16beta-methyl) was dissolved in 100ml of dichloromethane. 200 ml of 7 mol/L hydrochloric acid and 20 g of paraformaldehyde were added, stirred for 24 hours, and the reaction was confirmed by TLC (eluent n-Hex:EA = 1:1 Rf = 0.4). When the reaction was complete, the organic layer was evaporated. Then recrystallized from methanol to obtain 6.7 g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-benzoxyl-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] (Pregna- Synthesis of 1,4-dien-3-one,9-fluoro-11beta-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)]).

6.7 g of the product of 1 (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methylenebis(oxy)]) was dissolved in tetrahydrofuran. dissolved in It was cooled at 0° C. and 1.2 g of sodium hydrogen (60%) was added. After stirring for 30 minutes, 2 g of benzyl bromide was dropped into the reaction solution and stirred overnight. The reaction was quenched with water and purified by silica gel column chromatography (eluent n-Hex:EA = 9:1 to 5:1) to give 1.1 g of product.

3. Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxyl, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl (Pregna- Synthesis of 1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl).

20 ml tetra It was dissolved in hydrofuran, cooled at 0°C, added with 20ml hydrofluoric acid, and stirred for 24 hours. The organic layer was extracted with ethyl acetate, and the organic solvent was evaporated to obtain 890 mg of product.

4. Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta benzoxyl, 17-hydroxy-10,13,16-beta-trimethyl (Pregna-1 ,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta benzoxy,17-hydroxy-10,13, 16-beta-trimethyl) synthesis.

890 mg of the product of 3 (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl) was added to 5 ml pididine. and after cooling at 0°C, 250 mg of methanesulfonyl chloride was added and the reaction was confirmed by TLC (eluent n -Hex:EA = 2:1, Rf = 0.31) until completion. After adding ethyl acetate to the residue and washing with water, the organic solvent was evaporated to give the crude product. It was dissolved directly in butanone, 2 g of anhydrous lithium chloride was added, and the mixture was stirred overnight at room temperature. The organic layer was evaporated and purified by silica gel column chromatography (eluent n-Hex:EA = 5:1) to obtain 120 mg of product. This is hereinafter referred to as the S002 compound or the compound of Formula 2.

[Formula 2]

The results of NMR analysis of the S002 compound (Formula 2) are as follows:

^1H NMR (500 MHz, CDCl ₃ ) δ 7.344 (m, 5H), 6.792 (d, 1H,J=13Hz), 6.258 (dd, 1H, J1=12.5Hz, J2=2Hz,), 6.079 (s, 1H), 4.743 (d, 1H, J=13.5Hz,), 4.500 (AB, 2H, J1=57.5Hz, J2=20Hz),4.313 (d, 1H, J=13.5Hz), 4.03 (dm,1H, J=11.5Hz), 1.406 (s, 3H), 1.184 (s, 3H), 1.148 (d, 2H, J=9Hz),

LC-MS (ESI) m/z: 501.0 [M+H] ⁺

HPLC purity: 98.09%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=50:50

Production Example 3 (S003)

Synthesis was performed according to Scheme 3 below.

[Scheme 3]

1. Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enbis(oxy)]( Synthesis of Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis(oxy)]).

10 g of betamethasone (Pregna-1,4-diene-3,20-dione, 9-fluoro-11beta,17alpha,21-trihydroxy-16beta-methyl) was dissolved in 100ml of dichloromethane. 200 ml of 7 mol/L hydrochloric acid and 20 g of paraformaldehyde were added, stirred for 24 hours, and the reaction was confirmed by TLC (eluent n-Hex:EA = 1:1 Rf = 0.4). When the reaction was complete, the organic layer was evaporated. Then recrystallized from methanol to obtain 6.7 g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-16beta-methyl-17,20:20,21-bis[methylene Bis(oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy )]) Synthesis of.

6.7 g of the product of 1 (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methylenebis(oxy)]) was dissolved in tetrahydrofuran. dissolved in It was cooled at 0° C. and 1.2 g of sodium hydrogen (60%) was added. After stirring for 30 minutes, 2 g of (4'-isopropyl)-benzyl bromide was dropped into the reaction solution and stirred overnight. The reaction was quenched with water and purified by silica gel column chromatography (eluent n-Hex:EA = 9:1 to 5:1) to give 0.95 g of product.

3. Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-isopropyl)-benzoxyl, 17-alpha, 21-dihydroxy-10,13 ,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-isopropyl)-benzoxy, 17-alpha, 21-dihydroxy-10,13, 16-beta-trimethyl) synthesis.

The product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) was dissolved in 20ml tetrahydrofuran, cooled at 0°C, and 20ml hydrofluoric acid was added and stirred for 24 hours. The organic layer was extracted with ethyl acetate, and the organic solvent was evaporated to obtain 560 mg of product.

4. Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta-(4'-isopropyl)-benzoxyl, 17-hydroxy-10,13, 16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta-(4'-isoproyl)-benzoxy,17-hydroxy-10,13,16 -beta-trimethyl) synthesis.

The product of 3 (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-isopropyl)-benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta -trimethyl) 560 mg was dissolved in 5 ml pididine, cooled at 0 ° C, and then 250 mg of methanesulfonyl chloride was added and the reaction was completed by TLC (eluent n -Hex:EA = 2: 1, Rf = 0.31). Confirmed. After adding ethyl acetate to the residue and washing with water, the organic solvent was evaporated to give the crude product. This was directly dissolved in butanone, 2 g of anhydrous lithium chloride was added, and stirred at room temperature for 18 hours. The organic layer was evaporated and purified by silica gel column chromatography (eluent n-Hex:EA = 5:1) to give 125 mg of product. This is hereinafter referred to as the S003 compound or the compound of Formula 3.

[Formula 3]

The results of NMR analysis of the S003 compound (Formula 3) are as follows:

^1H NMR (500 MHz, CDCl ₃ ) δ 7.18 (d, 1H, J=10Hz), 7.07 (m, 4H), 6.38 (d, 1H, J=10.2Hz), 4.43 (AB, 2H, J1=68Hz , J2=16Hz), 4.38 (m, 1H), 3.82 (AB, 2H, J1=52Hz, J2=15Hz), 2.84 (m, 2H), 1.56(s,3H),1.21 (d,6H, J= 17Hz), 1.165 (s, 3H), 1.15 (d, 3H, J=7.5Hz),

LC-MS (ESI) m/z: 543.0 [M+H] ⁺

HPLC purity: 97.13%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=50:50

Production Example 4 (S004)

Synthesis was performed according to Scheme 4 below.

[Scheme 4]

1. Pregn-4-ene-3,20-dione,10,13-dimethyl-17,21-[(1-methoxypropylidene)bis(oxy)](Pregn-4-ene-3,20- Synthesis of dione,10,13-dimethyl-17,21-[(1-methoxypropylidene)bis(oxy)]).

2 g of cortexolone (Pregna-4-ene-3-one-10,13-dimethyl-17alpha,21-dihydroxy) was dissolved in tetrahydrofuran. It was cooled at 0°C, and 20 ml of 2 mol/L hydrochloric acid was added to the reaction solution. It was stirred for 15 hours, the reaction was quenched with water, and purified by silica gel column chromatography (eluent n-Hex:EA = 0.4) to give 0.45 mg of product.

2. Pregn-4-ene-3,20-dione-10,13-dimethyl-21-chloro-17-hydroxy-17-propionate (Pregn-4-ene-3,20-dione-10, Synthesis of 13-dimethyl-21-chloro-17-hydroxy-17-propionate).

450 mg of the product of 1 (Pregn-4-ene-3,20-dione-10,13-dimethyl-17, 21-dihydroxy-17-propionate) was dissolved in 5 ml of pyridine. It was cooled at 0°C, 250 mg of methanesulfonyl chloride was added, and the reaction was checked by TLC (eluent n-Hex:EA = 2:1, Rf = 0.43) until completion.

After adding ethyl acetate and washing with water, the organic solvent was evaporated to give the crude product. This was directly dissolved in butanone, 2 g of anhydrous lithium chloride was added, and stirred at room temperature for 24 hours. The organic layer was evaporated and purified by silica gel column chromatography (eluent n-Hex:EA = 5:1) to obtain 189 mg of product.

3. Pregn-1,4-diene-3,20-dione-10,13-dimethyl-21-chloro-17-hydroxy-17-propionate (Pregn-1,4-diene-3,20- Synthesis of dionee-10,13-dimethyl-21-chloro-17-hydroxy-17-propionate).

189 mg of the product of 2 (Pregn-4-ene-3,20-dionee-10,13-dimethyl-21-chloro-17-hydroxy-17-propionate) was dissolved in 5 ml of 1,4-dioxane, and 120 mg of After adding DDQ, heat was applied and the reaction was checked by TLC (eluent n -Hex:EA = 2:1, Rf = 0.35) until complete. The organic solvent was evaporated, and the crude product was purified by SFC (CO ₂ :MeOH=98:2) to obtain 68 mg of product. This is hereinafter referred to as the S004 compound or the compound of Formula 4.

[Formula 4]

The results of NMR analysis of the S004 compound (Formula 4) are as follows:

^1H NMR (500 MHz, CDCl ₃ ) δ 7.05 (d, 1H, J=10.0Hz), 6.25 (dd, 1H, J1=10.0Hz, J2=1.5Hz), 6.093 (s, 1H), 4.132 (AB , 2H, J1=70.0Hz, J2=15Hz),2.91 (m, 1H), 2.495 (td,1H, J1=13.5Hz, J2=5.5Hz), 2.33-2.41(m,3H),1.24 (s, 3H), 1.143 (t, 3H), 0.757 (3, 3H),

LC-MS (ESI) m/z: 419.0 [M+H] ⁺

HPLC purity: 99.68%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=60:40

Preparation Example 5 (S005)

Synthesis was performed according to Scheme 5 below.

[Scheme 5]

11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl)-delta4,9-estradien-3-one-17beta-hydroxy-( 2'-chloro)acetate (11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl)-delta4,9-estradiene-3-one-17beta-hydroxy-(2 Synthesis of '-chloro)acetate).

2 g of mifepristone (11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl)-delta 4,9-estradiene-3-one-17beta-hydroxy) was added to 20 ml of dichloromethane. and 2 g of chloroacetyl chloride was added. The reaction was checked by TLC (eluent n-Hex:EA = 1:1, Rf = 0.52) until completion. The organic solvent was evaporated, and the product was purified in high-performance liquid phase (pre-HPLC acetonitrile: water = 50:50) to obtain 120 mg of the product. This is hereinafter referred to as the S005 compound or the compound of Formula 5.

[Formula 5]

The results of NMR analysis of the S005 compound (Formula 5) are as follows:

¹ H NMR (500 MHz, CDCl ₃ ) δ 7.03 (d, 2H, J=10.5 Hz), 6.68 (dm, 2H, J=9.0 Hz), 6.81 (d, 1H), 5.76 (s, 1H) , 4.36 (d, 1H, J=9.0Hz),3.997 (s, 2H), 2.93 (s, 6H), 191(s,3H),0.566 (s, 3H),

LC-MS (ESI) m/z: 430.0 [M+H] ⁺

HPLC purity: 98.19%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=50:50

Preparation Example 6 (S006)

Synthesis was performed according to Scheme 6 below.

[Scheme 6]

1. Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enbis(oxy)]( Synthesis of Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis(oxy)]).

10 g of betamethasone (Pregna-1,4-diene-3,20-dione, 9-fluoro-11beta,17alpha,21-trihydroxy-16beta-methyl) was dissolved in 100ml of dichloromethane. 200 ml of 7 mol/L hydrochloric acid and 20 g of paraformaldehyde were added, stirred for 24 hours, and the reaction was confirmed by TLC (eluent n-Hex:EA = 1:1 Rf = 0.4). When the reaction was complete, the organic layer was evaporated. Then recrystallized from methanol to obtain 6.7 g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxyl-16beta-methyl-17,20:20,21-bis[methylenebis (oxy)](Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy) ]) Synthesis of.

6.7 g of the product of 1 (Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, 21-bis[methylenebis(oxy)]) was dissolved in tetrahydrofuran. dissolved in It was cooled at 0° C. and 1.2 g of sodium hydrogen (60%) was added. After stirring for 30 minutes, 2 g of benzyl bromide was dropped into the reaction solution and stirred for 24 hours. The reaction was quenched with water and purified by silica gel column chromatography (eluent n-Hex:EA = 9:1 to 5:1) to give 3.1 g of product.

3. Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-chloro)-benzoxyl, 17-alpha, 21-dihydroxy-10,13, 16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,9-fluoro-11-beta-(4'-chloro)-benzoxy,17-alpha,21-dihydroxy-10,13,16 -beta-trimethyl) synthesis.

The product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) was dissolved in 20ml tetrahydrofuran, cooled at 0°C, and 20ml hydrofluoric acid was added and stirred for 14 hours. The organic layer was extracted with ethyl acetate, and the organic solvent was evaporated to obtain 2.6 g of product.

4. Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta (4'-chloro) -benzoxyl-16-beta-methyl-17-hydroxy ( Synthesis of Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro)-benzoxy-16-beta-methyl-17-hydroxy).

The product of 3 (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta-(4'-chloro)-benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta -trimethyl) 2.6 g was dissolved in 5 ml pididine, cooled at 0 ° C, and then 700 mg of methanesulfonyl chloride was added and the reaction was completed by TLC (eluent n-Hex:EA = 2: 1 Rf = 0.35). Confirmed. After adding ethyl acetate to the residue and washing with water, the organic solvent was evaporated to give the crude product. It was dissolved directly in butanone, 2 g of anhydrous lithium chloride was added, and stirred for 24 hours. The organic layer was evaporated and purified by silica gel column chromatography (eluent n-Hex:EA = 5:1) to give 1.9 g of product.

5. Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta (4'-chloro-benzoxyl)-16-beta-methyl-17-hydroxy pro Synthesis of cionate (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro-benzoxy)-16-beta-methyl-17-hydroxy propionate) .

Product of 4 (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro)-benzoxy-16-beta-methyl-17-hydroxy) 1.9g 5 ml of trifluoroacetic acid and 5 ml of propionic anhydride were mixed, 50 mg of p-toluenesulfonic acid was added, and the reaction was heated at 50 °C until the reaction was completed by TLC (eluent n-Hex:EA = 2: 1 Rf = 0.47). The crude product was purified by silica gel column chromatography (eluent n-Hex:EA = 4:1) to obtain 130 mg of product. This is hereinafter referred to as the S006 compound or the compound of Formula 6.

[Formula 6]

The results of NMR analysis of the S006 compound (Formula 6) are as follows:

¹ H NMR (500 MHz, CDCl ₃ ) δ 7.34 (d, 2H, J=8.5 Hz), 7.25 (d, 2H, J=8.5 Hz), 6.8 1 (d, 1H, J=10.5 Hz), 6.32 ( dd, 1H, J1=10.5Hz, J2=2Hz), 6.12(s, 1H),4.69 (d, 1H, J=11Hz), 4.32 (d, 1H, J=11Hz), 4.09 (m, 1H), 4.069 (d, 1H, J=13.5Hz),3.904(d, 1H, J=14.0Hz),2.39(q, 2H, J=8.0Hz), 1.41(s,3H),1.38(d,3H, J =7.5Hz), 1.16 (t, 3H, J=7.5Hz), 1.000 (s, 3H),

LC-MS (ESI) m/z: 591.0 [M+H] ⁺

HPLC purity: 99.39%, Waters e2695, Xbridge C18, 210 nm, acetonitrile:water=50:50

Example 2: Confirmation of anti-cancer effect of novel compounds

preparation example

For use in the experiment, non-small-cell lung carcinoma (NSCLC) A549 cells were purchased from the Korea Cell Line Bank and supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin-strepto In a DMEM medium (HyClone Laboratories, USA) supplemented with mycin and 1% HEPES, they were cultured under 5% CO ₂ and 37°C conditions. In addition to the compound prepared in Example 1 of the present invention, dimethyl sulfoxide (DMSO) as a negative control and mifepristone (RU486) of Formula B as a positive control were purchased from Sigma-Aldrich (St. Louis, USA) and used.

[Formula B]

Example 2-1. Confirmation of cancer cell survival inhibitory effect of new compound

After seeding 1×10 ⁴ cells in a 96-well plate in order to confirm the cancer cell survival inhibitory effect of the novel compounds, they were cultured for 24 hours in DMEM medium containing 1 μM, 10 μM, 25 μM, or 50 μM of each compound. Then, the supernatant was removed, washed with PBS, further cultured in serum-free DMEM medium containing MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) for 4 hours, and the medium was removed. , 100 μl of DMSO was added and stirred for 1 minute to dissolve the purple foam cup. It was measured at 595 nm. The results are shown in Figure 1.

As a result of the experiment, it was confirmed that the new compounds S001 to S006 had an effect of inhibiting cancer cell survival in proportion to the concentration. In particular, S005 showed an excellent cancer cell survival inhibitory effect at a low concentration of 1 μM or less, and S001 to S003 showed an excellent cancer cell survival inhibitory effect at a concentration of 25 μM or more compared to the positive control RU486. At concentrations of 50 μM or more, the cancer cell survival inhibitory effect of S002 and S003 was very remarkable.

Example 2-2. Confirmation of cancer cell tumor formation inhibitory effect of new compound

A colony formation experiment was performed to confirm the inhibitory effect of the new compound on cancer cell tumorigenesis. A tumor is a general term for all abnormally generated masses, and cancer cells are characterized by forming a colony by dividing and proliferating indefinitely beyond normal division control. To this end, the cells were seeded in a 12-well plate at a very low concentration (3×10 ³ ) and cultured in DMEM medium containing 1 nM, 10 nM, 100 nM, 1 μM, 10 μM, 25 μM, or 50 μM of each compound for 2 weeks. Thereafter, the degree of colony formation was observed using a crystal violet staining method to confirm the anticancer effect. Specifically, after removing the supernatant and washing with PBS, the remaining cells were stained with a crystal violet solution (20% methanol and 0.5% crystal violet) for 10 minutes at room temperature. After washing three times and incubating for solubilization in 1% SDS solution, absorbance at 570 nm wavelength was measured. The results are shown in FIG. 2 .

As a result of the experiment, S001 to S005 had an excellent cancer cell tumor formation inhibitory effect at a concentration of 1 μM. In particular, S005 showed a tumor formation inhibitory effect of 50% or more compared to the negative control, DMSO, at a concentration of 100 nM, and it was found that the effect was remarkable at a low concentration, similar to the cancer cell survival inhibitory effect of Example 2-1. Positive controls, RU486 and S006, were found to have significant tumor formation inhibitory effects at concentrations of 25 μM or higher.

Example 2-3. Confirmation of cancer cell invasion inhibitory effect of new compound

In order to confirm the cancer metastasis inhibitory effect of the new compound, the cancer cell invasion inhibitory effect was confirmed through a soft agar assay. Specifically, after putting 50 μl of DMEM medium containing 0.7% agar in a 12-well plate and solidifying (bottom agar), 300 μl of DMEM medium containing 3×10 ³ cells and 0.35% agar was put on the solidified agar medium and solidified ( top agar). After hardening, 250 μl of DMEM medium containing 1 μM or 10 μM of each compound was added and cultured in a CO ₂ incubator for about 2 weeks, and the number of colonies formed on 0.7% bottom agar was determined. The results are shown in Figure 3.

As a result of the experiment, the compounds S002, S004, and S006 had excellent inhibitory effects on cancer cell invasion even at a concentration of 1 nM. S003 showed a significant invasion inhibitory effect at a concentration of 10 nM, and S001 and S005 showed a similar invasion inhibitory effect as the positive control RU486. However, all tested compounds had a significant cancer cell invasion inhibitory effect compared to the negative control, DMSO.

Having described specific parts of the present invention in detail above, it is clear that these specific techniques are merely preferred embodiments for those skilled in the art, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (8)

A compound selected from a derivative of a compound represented by Formula A below, a pharmaceutically acceptable salt thereof, an optical isomer, a hydrate, and a solvate thereof,
[Formula A]

A compound of which the derivative of the compound represented by Formula A is a compound represented by any one of the following Formulas 1 to 6:
[Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

. delete delete delete delete delete delete delete

Images