KR102604396B1 - Novel compound and their uses - Google Patents

Abstract

The present invention relates to novel mifepristone derivative compounds and various uses thereof.
In order to develop an effective cancer treatment, it must be able to simultaneously inhibit the growth and metastasis of cancer cells. Generally, cancer growth refers to the infinite proliferation characteristics of cancer cells, and cancer metastasis refers to the characteristics of cancer cells separating from the primary site and moving to another location. Therefore, cancer growth and cancer metastasis require approaches from different mechanisms.
Since the compound provided in the present invention can simultaneously inhibit cancer proliferation and metastasis, it is expected to be actively used in the medical and health fields.

Description

Novel compound and their uses}

The present invention relates to novel mifepristone derivative compounds 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 most common cause of death.

In order to develop an effective cancer treatment, it must be able to simultaneously inhibit the growth and metastasis of cancer cells. In general, cancer growth refers to the infinite proliferation characteristic of cancer cells. Normal cells either complete the number of divisions programmed in their genes in advance, or stop dividing when there is insufficient space for new cells. However, cancer cells have failed to regulate the function that controls division, and when they run out of space to expand themselves, they secrete a protease (matrix metalloprotease) that decomposes the cell matrix, eliminating the surrounding space to create space for themselves. expand On the other hand, cancer metastasis refers to the characteristic of cancer cells separating from the primary site and moving to another location. To achieve this, ① cancer cells must be eliminated from the primary site (cancer tissue), ② they must invade and pass through the tissue walls that make up blood vessels or lymph vessels, and ③ they must attach to and fuse with the matrix of the moved tissue in order to settle in a new location. , an approach from a different mechanism from cancer growth is needed.

Therefore, the present invention was devised to solve the above problems, and relates to a novel compound capable of simultaneously inhibiting cancer proliferation and metastasis, and its pharmaceutical use.

One object of the present invention is to provide a new compound that can prevent, improve or treat cancer.

Another object of the present invention is to provide a composition that can prevent, improve or treat cancer, including the above-mentioned compounds.

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

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

DETAILED DESCRIPTION OF THE INVENTION Various embodiments described herein are described below with reference to the drawings. In the following description, various specific details, such as specific forms, compositions, and processes, are set forth in order to provide a thorough understanding of the invention. However, certain embodiments may be practiced without one or more of these specific details or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques are not described in specific detail so as not to unnecessarily obscure the 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. Accordingly, the phrases “in one embodiment” or “an embodiment” expressed in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, particular features, shapes, compositions, or properties may be combined in any suitable way in one or more embodiments.

Unless there is a special definition in the specification, all scientific and technical terms used in the specification have the same meaning as commonly understood by a person skilled in the art in the technical field to which the present invention pertains.

In the present invention, “cancer” is characterized by uncontrolled cell growth. Due to this abnormal cell growth, a cell mass called a tumor is formed, which infiltrates surrounding tissues and, in severe cases, may metastasize to other organs of the body. means that Academically, it is also called a neoplasm. Cancer is an incurable chronic disease that cannot be fundamentally cured even when treated with surgery, radiation, and chemotherapy in many cases, causes pain to the patient, and ultimately leads to death. There are many causes of cancer, but internal factors are the cause. It is divided into and external factors. Although the exact mechanism by which normal cells are transformed into cancer cells has not been identified, it is known that a significant number of cancers are caused 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 them is broken 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, laryngeal cancer, oral cancer, and head and neck cancer. , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, etc., and is limited to types of cancer in which cancer progression, such as tumor differentiation and/or proliferation, is dependent on the cancer cells and/or cancer stem cells described in the present invention. It doesn't work. At this time, the above cancer stem cell (CSC) refers to cancer cells in a comprehensive sense that have self-renewal ability or differentiation ability, which are unique abilities of stem cells. Cancer stem cells make up 0.1 to 5% of cancer tissues. It is presumed that it exists in between. The cancer stem cells are grown under normal tumor growth conditions (the "normal tumor growth conditions" refer to a state in which there are sufficient nutrients (glucose) necessary for cell growth and the growth conditions of the tumor microenvironment are abundant, so there is no cell stress.) Unlike general cancer cells, they have the characteristic of proliferating slowly. In addition, they may maintain a dormant state and have resistance to anticancer drugs. Unlike normal tumor cells, the expression of transcriptional regulators such as PGC-1α is controlled, so the functions of major metabolic regulators are different from those of normal cancer cells. There are different characteristics. It comprehensively refers to cells with the ability to invade and/or metastasize by acquiring resistance to cell death (apoptosis) under nutrient starvation conditions through these different metabolic controls and the control of cell signaling systems linked thereto. However, as long as it is a cell that can differentiate into a general cancer cell, it is not limited thereto.

In the present invention, “virtual screening” is one of the numerous procedures that go through during the process of developing a new drug. More specifically, it refers to the procedure of discovering effective substances and lead substances through screening of compounds selected from basic research and confirming candidate substances. says This can reduce the time and cost required to develop new drugs, and after the target disease is selected, effective substances for new drugs targeting it can be discovered, and drug efficacy can be predicted by computer using 3D reconstructed gene protein information. It is also possible to selectively search for suitable substances. Virtual screening technology can be used not only for library creation but also for active screening of actually existing libraries and specific action points. However, if it is a general and universal screening technology, it is not limited to this.

In the present invention, a compound of the following formula (A) was identified as a candidate anticancer agent through virtual screening as described above, and its derivative 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 Formula A may be a new compound synthesized in the present invention and may be represented by the following Formulas 1 to 6.

[Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

The formula 1 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, according to Scheme 1 described in the present invention. 21-bis[methyl-enebis(oxy)](Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-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)-benzoxy,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 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'- It can be named chloro)-benzoxy-16-beta-methyl-17-hydroxy).

The formula 2 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, according to Scheme 2 described in the present invention. 21-bis[methyl-enebis(oxy)](Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-benzoxy-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 benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl ( Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl). Preparation comprising: Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta benzoxy, 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).

Formula 3 is (a) pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, according to Scheme 3 described in the present invention. 21-bis[methyl-enebis(oxy)](Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-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)-benzoxy, 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, Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11- Beta-(4'-isopropyl)-benzoxy, 17-hydroxy-10,13,16-beta-trimethyl (Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro- It can be named 11-beta-(4'-isoproyl)-benzoxy,17-hydroxy-10,13,16-beta-trimethyl).

The above formula 4 is (a) preg-4-en-3,20-dione,10,13-dimethyl-17,21-[(1-methoxypropylidene)bis(oxy) according to Scheme 4 described in the present invention. )] (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- Preg-1,4-diene-3,20-dione-10 is manufactured by a manufacturing method comprising the step of synthesizing 10,13-dimethyl-21-chloro-17-hydroxy-17-propionate). ,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, 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).

The above formula (6) is (a) pregna-1,4-diene-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20, according to Scheme 6 described in the present invention. 21-bis[methyl-enebis(oxy)](Pregna-1,4-dien-3-one, 9-fluoro-11beta-hydroxy-16beta-methyl-17,20:20,21-bis[methyl-enebis (oxy)]); (b) Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-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)-benzoxy, 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-hyde. Steps to synthesize Roxy (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-benzoxy)-16 -beta-methyl-17-hydroxy propionate (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 above-described novel compound, a method for producing the same (synthetic method), and also provides a pharmaceutically acceptable salt of the compound. Said pharmaceutically acceptable salts may be salts generally regarded by those skilled in the art as being suitable for medical applications (e.g. because such salts are not harmful to the subjects to be treated with said salts), or salts within the respective treatment. It is a salt that causes acceptable side effects. Generally, the pharmaceutically acceptable salts are salts that are considered acceptable by regulatory authorities such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or the Pharmaceutical and Medical Devices Agency (PMDA) of the Japanese Ministry of Health, Labor and Welfare. . However, the present invention in principle also provides, for example, intermediates in the preparation of the compounds according to the invention or physiologically functional derivatives thereof, or pharmaceutically acceptable salts of the compounds according to the invention or physiologically functional derivatives thereof. As intermediates in the preparation of derivatives, also include salts of the compounds according to the invention which are not pharmaceutically acceptable as such. The salts include water-insoluble salts and, in particular, water-soluble salts.

In each case, the person skilled in the art will be able to determine whether a particular compound according to the invention or a physiologically functional derivative thereof is capable of forming salts, i.e. whether the compound according to the invention or a physiologically functional derivative thereof is capable of forming salts, e.g. For example, it is easy to determine whether it has a group that can bear a charge, such as an amino group, carboxylic acid group, etc.

Exemplary salts of the compounds of the invention are acid addition salts or salts with bases, especially pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases commonly used in pharmaceuticals, which are water-insoluble or especially water-soluble acid addition salts. It's salt. Depending on the substituents of the compounds of the invention, salts with bases may also be suitable. Acid addition salts include, for example, a solution of a compound of the 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). Illustrative examples of pharmaceutically acceptable salts include acetate, adipate, alginate, arginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, Bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, digluconate, dihydrochloride, dodecyl sulfate, 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, maleate, 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, Includes, but is not limited to, valerate.

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

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

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

Hereinafter, the compounds of the present invention refer to the compounds described above.

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

At this time, the diseases of interest include skeletal diseases, joint diseases, vascular diseases, blood diseases, skin diseases, kidney diseases, cardiovascular diseases, cranial nerve diseases, ophthalmic diseases, digestive diseases, urinary diseases, gynecological diseases, respiratory diseases, infectious diseases, and allergies. It may be a disease, a rheumatic disease, an autoimmune disease, a disease caused by transplantation or a side effect of transplantation, or a proliferative disease (tumor).

The present invention provides a pharmaceutical composition comprising as an active ingredient a compound selected from the compounds of the present invention, pharmaceutically acceptable salts, crystalline forms, co-crystals, optical isomers, derivatives, hydrates, and solvates 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 creation or growth of cancer or improve symptoms of cancer invasion, metastasis and recurrence.

In the present invention, cancer as an object for improvement or treatment with a pharmaceutical composition containing the compound of the present invention as an active ingredient includes liver cancer, biliary tract 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 one or more types 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 a capsule, tablet, granule, injection, ointment, powder, or beverage, and the pharmaceutical composition may be intended for human subjects.

The pharmaceutical composition of the present invention is not limited to these, but can be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, external preparations, suppositories, and sterile injection solutions according to conventional methods. You can. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. for oral administration, and buffers, preservatives, and analgesics for injections. Topics, solubilizers, isotonic agents, stabilizers, etc. can be mixed and used, and for topical administration, bases, excipients, lubricants, preservatives, etc. can be used. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing it with a pharmaceutically acceptable carrier as described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injections, it can be manufactured in the form of unit dosage ampoules or multiple dosage forms. there is. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release preparation, etc.

Meanwhile, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil may be used. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be additionally 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, enteral, and topical. , sublingual or rectal. Oral or parenteral administration is preferred. The parenteral administration includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention can also be administered in the form of a suppository for rectal administration.

The pharmaceutical composition of the present invention may vary depending on several factors, including the activity of the specific compound used, age, body weight, general health, gender, diet, administration time, administration route, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated. It may vary, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, route and period of administration, but may be appropriately selected by a person skilled in the art, and may range from 0.0001 to 50 mg/day. It can be administered in kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered several times. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

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

In addition, the present invention provides a compound selected from the compounds of the present invention, pharmaceutically acceptable salts, crystalline forms, co-crystals, optical isomers, derivatives, hydrates, and solvates thereof to subjects in need of administration (e.g., humans). Provided is a method for preventing, improving, or treating cancer, comprising administering a pharmaceutical composition containing the ingredient in a pharmaceutically effective amount. At this time, “administration” means providing a given compound of the present invention to a subject by any appropriate method.

Specific details regarding cancer, pharmaceutical compositions, and administration routes in the method for preventing, improving, or treating cancer are described in the above “Compound of the present invention, pharmaceutically acceptable salts, crystalline forms, co-crystals, optical isomers, and derivatives thereof. , hydrate, and solvate. This overlaps with what was described in “Pharmaceutical composition containing a compound selected from hydrate, and solvate as an active ingredient,” and is omitted to prevent excessive complexity of the specification.

In the present invention, the “subject” requiring the administration may include both mammals and non-mammals. Here, examples of such mammals include humans, non-human primates, such as chimpanzees, and other ape and monkey species; Livestock animals such as cattle, horses, sheep, goats, and pigs; Domesticated animals such as rabbits, dogs and cats; Laboratory animals may include, but are not limited to, rodents such as rats, mice, and guinea pigs. Additionally, 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 reduction and/or alleviation of the signs, symptoms or causes of a disease, or any There may be other desirable changes. For example, an "effective amount" for therapeutic use is the amount of a compound disclosed in the present invention that is required to provide a clinically significant reduction in the 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 the amount at which the active substance has a therapeutic effect.

The present invention also provides a food composition for preventing or alleviating cancer containing a compound selected from the compounds of the present invention, food acceptable salts, crystalline forms, co-crystals, optical isomers, derivatives, hydrates, and solvates thereof. to provide.

The food composition herein is used in various ways for the purpose of the present invention, that is, for the prevention or improvement of cancer, and the food composition containing the compound of the present invention is used in various foods, such as beverages, gum, tea, It can be manufactured in the form of vitamin complexes, powders, granules, tablets, capsules, cookies, rice cakes, bread, etc. When the composition of the present invention is included in a food composition, it can be added in an amount of 0.1 to 100% of the total weight. Here, when the food composition is manufactured in the form of a beverage, there are no particular limitations other than containing the food composition in the indicated ratio, and various flavoring agents or natural carbohydrates can be contained as additional ingredients like ordinary beverages. That is, natural carbohydrates include common sugars such as monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, 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 (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). In addition, the present invention The food composition includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, It may contain stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These components can 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 in 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 cancer proliferation and metastasis, it is expected to be actively used in the medical and health fields.

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

The inventors of the present invention conducted virtual screening of various candidate substances to discover a substance for treating cancer cells and/or cancer stem cells with anticancer activity. As a result, they derived a compound of the following formula A as a candidate substance for anticancer use 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 refers to the USP test conditions for clobetasol propionate using a 4.6mm x 15cm column and C18 packing (pore size 5um). did.

Manufacturing 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)]).

10g 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. Afterwards, it was recrystallized with methanol to obtain 6.7g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-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) ]).

6.7g of product 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. This 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 added to 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)-benzoxy, 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.

Product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) 3.1g was dissolved in 20ml tetrahydrofuran, cooled at 0°C, 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)-benzoxy-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).

Products 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) 2.6g was dissolved in 5ml phycidine, cooled at 0°C, 700mg of methanesulfonyl chloride was added, and the reaction was monitored by TLC (eluent n-Hex:EA = 2:1 Rf=0.35) until the reaction was complete. Confirmed. Ethyl acetate was added to the residue, washed with water, and the organic solvent was evaporated to obtain the crude product. This 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 obtain 1.9 g of product. This is hereinafter referred to as the S001 compound, or the compound of Formula 1.

[Formula 1]

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

^1H NMR (500 MHz, CDCl ₃ ) δ 7.343(d, 2H, J=10.5Hz), 7.265(d, 2H, J=10.5Hz), 6.82(d, 1H, J=12.5Hz), 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, 210nm, acetonitrile:water=50:50

Manufacturing 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)]).

10g 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. Afterwards, it was recrystallized with methanol to obtain 6.7g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-benzoxy-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)]).

6.7g of product 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. This 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 added to 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 benzoxy, 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).

1.1 g of the product of 2 (Pregna-1, 4-dien-3-one,9-fluoro-11beta-benzoxy-16beta-methyl-17,20:20,21-bis[methyl enebis(oxy)]) was added to 20 ml of tetramer. It was dissolved in hydrofuran, cooled at 0°C, 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 890 mg of product.

4. Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-beta benzoxy, 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).

890 mg of product 3 (Pregna-1,4-diene-3,20-dione, 9-fluoro-11-beta benzoxy, 17-alpha, 21-dihydroxy-10,13,16-beta-trimethyl) was mixed with 5 ml phydidine. After dissolving in and 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 the reaction was completed. Ethyl acetate was added to the residue, washed with water, and the organic solvent was evaporated to obtain the crude product. This was dissolved directly in butanone, 2 g of anhydrous lithium chloride was added, and stirred at room temperature overnight. 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 NMR analysis results 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, 210nm, acetonitrile:water=50:50

Manufacturing 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)]).

10g 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. Afterwards, it was recrystallized with methanol to obtain 6.7g 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.

6.7g of product 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. This 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 added to 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)-benzoxy, 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, Synthesis of 16-beta-trimethyl).

Product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-isopropyl)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) 0.95g was dissolved in 20ml tetrahydrofuran, cooled at 0°C, 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)-benzoxy, 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.

Products 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 phydidine, cooled at 0°C, 250 mg of methanesulfonyl chloride was added, and the reaction was carried out by TLC (eluent n -Hex:EA = 2:1, Rf=0.31) until the reaction was complete. Confirmed. Ethyl acetate was added to the residue, washed with water, and the organic solvent was evaporated to obtain the crude product. This was dissolved directly 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 obtain 125 mg of product. This is hereinafter referred to as the S003 compound, or the compound of Formula 3.

[Formula 3]

The NMR analysis results 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, 210nm, 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)]).

2g of Cortexolone (Pregna-4-ene-3-one-10,13-dimethyl-17alpha,21-dihydroxy) was dissolved in tetrahydrofuran. This was cooled at 0°C, and 20 ml of 2 mol/L hydrochloric acid was added to the reaction solution. This was stirred for 15 hours, and 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 product 1 (Pregn-4-ene-3,20-dione-10,13-dimethyl-17, 21-dihydroxy-17-propionate) was dissolved in 5 ml of pyridine. This was cooled at 0°C, 250 mg of methanesulfonyl chloride was added, and the reaction was confirmed by TLC (eluent n-Hex:EA = 2:1, Rf = 0.43) until completion.

After adding ethyl acetate and washing with water, the organic solvent was evaporated to give the crude product. This was dissolved directly 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 product 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 confirmed by TLC (eluent n -Hex:EA = 2:1, Rf = 0.35) until completion. 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 NMR analysis results 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, 210nm, acetonitrile:water=60:40

Production 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).

2g of mifepristone (11beta-[4'-(N,N'-dimethylamino)phenyl]-17alpha-(prop-1-ynyl)-delta 4,9-estradiene-3-one-17beta-hydroxy) was mixed with 20ml of dichloromethane. was dissolved in and 2g of chloroacetyl chloride was added. The reaction was confirmed 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 product. This is hereinafter referred to as the S005 compound, or the compound of Formula 5.

[Formula 5]

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

^1H NMR (500 MHz, CDCl ₃ ) δ 7.03(d, 2H, J=10.5Hz), 6.68(dm, 2H, J=9.0Hz), 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, 210nm, acetonitrile:water=50:50

Production Example 6 (S006)

Synthesis was performed according to Scheme 6 below.

[Scheme 6]

1. Pregna-1,4-diene-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)]).

10g 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. Afterwards, it was recrystallized with methanol to obtain 6.7g of product.

2. Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-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) ]).

6.7g of product 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. This 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 added to 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)-benzoxy, 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.

Product of 2 (Pregna-1,4-dien-3-one,9-fluoro-11beta-(4'-chloro)-benzoxy-16beta-methyl-17,20:20,21-bis[methylenebis(oxy)] ) 3.1g was dissolved in 20ml tetrahydrofuran, cooled at 0°C, 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)-benzoxy-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).

Products 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) 2.6g was dissolved in 5ml pididine, cooled at 0°C, 700mg of methanesulfonyl chloride was added, and the reaction was performed by TLC (eluent n-Hex:EA = 2:1 Rf=0.35) until the reaction was complete. Confirmed. Ethyl acetate was added to the residue, washed with water, and the organic solvent was evaporated to obtain the crude product. This 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 obtain 1.9 g of product.

5. Pregna-1,4-diene-3,20-dione,21-chloro-9-fluoro-11-beta(4'-chloro-benzoxy)-16-beta-methyl-17-hydroxy pro Synthesis of cypionate (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 Mix 5 ml of trifluoroacetic acid and 5 ml of propionic anhydride, add 50 mg of p-toluenesulfonic acid, and heat at 50°C until the reaction is complete. 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 NMR analysis results of the S006 compound (Formula 6) are as follows:

¹ H NMR (500 MHz, CDCl ₃ ) δ 7.34(d, 2H, J=8.5Hz), 7.25(d, 2H, J=8.5Hz), 6.8 1(d, 1H, J=10.5Hz), 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, 210nm, acetonitrile:water=50:50

Example 2: Confirmation of anticancer effect of new compound

Preparation example

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

[Formula B]

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

To confirm the effect of the new compound on suppressing cancer cell survival, 1 × 10 ⁴ cells were seeded in a 96-well plate and cultured for 24 hours in DMEM medium containing 1 μM, 10 μM, 25 μM, or 50 μM of each compound. Afterwards, the supernatant was removed, washed with PBS, and cultured for an additional 4 hours with serum-free DMEM medium containing MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide), and the medium was removed. , 100㎕ of DMSO was added and stirred for 1 minute to dissolve the purple foam. This 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 have an effect of suppressing the survival of cancer cells in proportion to their concentration. In particular, S005 showed an excellent effect in suppressing cancer cell survival at a low concentration of 1 μM or less, and S001 to S003 showed an excellent effect in suppressing cancer cell survival at a concentration of 25 μM or more compared to RU486, a positive control. At concentrations above 50 μM, the cancer cell survival inhibitory effect of S002 and S003 was very significant.

Example 2-2. Confirmation of the inhibitory effect of new compounds on cancer cell tumor formation

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

As a result of the experiment, S001 to S005 had an excellent effect in suppressing cancer cell tumor formation starting at a concentration of 1 μM. In particular, S005 already showed a tumor formation inhibition effect of more than 50% compared to DMSO, a negative control, at a concentration of 100 nM, and similar to the cancer cell survival inhibition effect in Example 2-1, the effect was significant at low concentrations. The positive controls, RU486 and S006, were shown to have a significant tumor formation inhibition effect at concentrations of 25μM or higher.

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

In order to confirm the effect of the new compound on suppressing cancer metastasis, the effect of suppressing cancer cell invasion was confirmed through soft agar assay. Specifically, 50 ㎕ of DMEM medium containing 0.7% agar was added to a 12-well plate and solidified (bottom agar), and then 300 ㎕ of DMEM medium containing 3 × 10 ³ cells and 0.35% agar was placed on top of the solidified agar (bottom agar). 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, compounds S002, S004, and S006 had excellent cancer cell invasion inhibition effects even at a concentration of 1 nM. S003 showed a significant invasion inhibition effect at a concentration of 10nM, and S001 and S005 showed a similar level of invasion inhibition effect as the positive control RU486. However, all tested compounds had a significant cancer cell invasion inhibition effect compared to the negative control, DMSO.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (12)

A pharmaceutical composition for the prevention or treatment of cancer comprising as an active ingredient a compound selected from the group consisting of a compound represented by the following formula (5), a pharmaceutically acceptable salt, crystalline form, co-crystal, optical isomer, hydrate, and solvate thereof:
[Formula 5]
.
delete delete delete delete delete delete delete According to clause 1,
The above cancers include liver cancer, biliary tract 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, and multiple myeloma. A pharmaceutical composition selected from the group consisting of leukemia, lymphoma, squamous cell cancer, kidney cancer, urethral cancer, bladder cancer, head and neck cancer, brain cancer, and central nervous system cancer.
According to clause 1,
A pharmaceutical composition wherein the cancer treatment inhibits the proliferation of cancer or inhibits the metastasis of cancer.
According to clause 1,
The pharmaceutical composition may be in oral dosage forms such as powders, granules, capsules, tablets, powders, or aqueous suspensions; Topical preparation; suppositories; Or, a pharmaceutical composition prepared in the form of a sterile injectable solution.
A food composition for preventing or improving cancer comprising as an active ingredient a compound selected from the group represented by the following formula (5), pharmaceutically acceptable salts, crystalline forms, co-crystals, optical isomers, hydrates, and solvates thereof:
[Formula 5]
.