WO2025178159A1 - Novel thioamide compounds, production method therefor, and uses thereof - Google Patents

Abstract

The present invention relates to a novel thioamide compound, a production method therefor, and uses thereof. The novel thioamide compound, stereoisomers thereof, solvates, or pharmaceutically acceptable salts of the present invention not only exhibit anticancer efficacy against various carcinomas, but also have excellent anticancer activity against anticancer drug-resistant cancers, and thus can be used for the prevention or treatment of various types of cancers.

Description

Novel thioamide compounds, methods for producing them, and uses thereof

The present invention relates to a novel thioamide compound, a method for producing the same, and a use thereof.

Cancer is a leading cause of death worldwide, and in Korea, it is the leading cause of death, with 161.1 deaths per 100,000 people as of 2022. While the five-year survival rate for cancer patients is increasing due to increased early diagnosis and improved treatments, cancer incidence and mortality rates are also on the rise, with cancer deaths in Korea in particular experiencing a significant increase. The global anticancer drug market is expected to grow rapidly, from $193 billion in 2022 to $377 billion in 2027, at a compound annual growth rate of 13-16%.

For decades, bioactive compounds derived from microorganisms have been widely used as clinical medicines or as a source for new drug development. However, in the mid-1980s, the discovery of new compounds from nature declined and the rediscovery of already known compounds became increasingly difficult. This led to rapid advancements in organic synthesis techniques capable of simultaneous synthesis of multiple organic compounds. However, developing new drugs from known compounds proved inefficient, and existing drugs were experiencing adverse effects, such as the development of pathogenic bacteria resistant to existing drugs and the development of anticancer-resistant cancer cells. These shortcomings have led to a surge in research on microbial-based natural products.

Natural products are becoming increasingly important as a source of biologically active drug candidates. Natural products account for 23.5% of all approved and used drugs worldwide over the past 40 years, and synthetic drugs with pharmacokinetic groups derived from natural products account for approximately 14% of the total, highlighting the importance of research on natural product medicines.

Accordingly, the inventors of the present invention isolated and purified a novel thioamide compound from a microorganism and confirmed its excellent anticancer effect, thereby completing the present invention.

One aspect provides a compound represented by chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.

Another aspect provides a Streptomyces sp. GC5 strain (Accession No.: KCTC15723BP) producing a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.

Another aspect provides a method for preparing a compound of formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, comprising the steps of: 1) culturing a strain of Streptomyces sp. GC5 (Accession No.: KCTC15723BP); and 2) isolating a compound of formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof from the strain, a lysate thereof, a culture medium thereof, or an extract of the culture medium.

Another aspect provides a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.

Another aspect provides a method for preventing or treating cancer, comprising administering to a subject a pharmaceutical composition for preventing or treating cancer.

Another aspect provides a food composition for preventing or improving cancer, comprising a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.

Another aspect provides the use of a compound represented by the above formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating cancer.

Another aspect provides the use of a compound represented by the above formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, for use in preventing or treating cancer.

One aspect is to provide a compound represented by the formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In the above chemical formula 1,

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R 7 , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , and ^{R 18 are} each independently hydrogen, a hydroxy group, an amino group, a halogen group, a ketone group, a cyano group, -C(=O)R _a , -C(=O)OR _a , -OCO(OR _a ), -C=N(R _a ), -SR _a , -S(=O)R _a , -S(=O) ₂ R _a , -PR _a , A substituted or unsubstituted C ₁ to C ₂₀ alkyl group, a substituted or unsubstituted C ₁ to C ₂₀ alkoxy group, a substituted or unsubstituted C ₂ to C ₂₀ alkenyl group, a substituted or unsubstituted C ₂ to C ₂₀ alkynyl group, a C ₂ to C ₂₀ alkylene oxide group, a substituted or unsubstituted C ₃ to C ₃₀ cycloalkyl group, a substituted or unsubstituted C ₆ to C ₃₀ aryl group, a substituted or unsubstituted C ₆ to C ₃₀ aryloxy group, a substituted or unsubstituted C ₆ to C ₃₀ heteroaryl group, or a combination thereof,

Here, R _a is hydrogen, a substituted or unsubstituted C1-C20 alkyl group, a substituted or unsubstituted C1-C20 alkoxy group, a substituted or unsubstituted C2-C20 alkenyl group, a substituted or unsubstituted C2-C20 alkynyl group, a substituted or unsubstituted C2-C20 alkylene oxide group, a substituted or unsubstituted C3-C30 cycloalkyl group, a substituted or unsubstituted C3-C30 heterocycloalkyl group, a substituted or unsubstituted C3-C30 cycloalkenyl group, a substituted or unsubstituted C3-C30 heterocycloalkenyl group, a substituted or unsubstituted C6-C30 aryl group, a substituted or unsubstituted C6-C30 aryloxy group, a substituted or unsubstituted C6-C30 arylthio group, a substituted or unsubstituted C6-C30 heteroaryl group, or any of these. It's a combination.

In one embodiment, R ¹¹ of the above chemical formula 1 may be hydrogen or a hydroxy group.

In one embodiment, at least one of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹² and R ¹³ of the above chemical formula 1 may be a substituted or unsubstituted C ₁ to C ₁₀ alkyl group, and specifically, may be a substituted or unsubstituted C ₁ to C ₅ alkyl group.

In one embodiment, at least one of R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁰ , R ¹² and R ¹³ of the chemical formula 1 may be a substituted or unsubstituted methyl group, and specifically, R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁰ , R ¹² and R ¹³ may be a methyl group.

In one embodiment, at least one of R ² and R ⁹ of the above chemical formula 1 may be a substituted or unsubstituted isopropyl group, and specifically, R ² and R ⁹ may be an isopropyl group.

In one embodiment, R ¹⁶ of the above chemical formula 1 may be hydrogen or a hydroxy group, and specifically may be hydrogen.

In one embodiment, the compound represented by Chemical Formula 1 may be represented by Chemical Formula 2 or Chemical Formula 3:

[Chemical Formula 2]

[Chemical Formula 3]

In one embodiment, the compound of formula 2 may be named Thiogochangamide A, and the compound of formula 3 may be named Thiogochangamide B.

The definitions of substituted substituents used in chemical formulas in this specification are as follows.

The term "hydroxy" as used in chemical formulas refers to the -OH functional group (hydroxyl group).

The term "amino" as used in chemical formulas refers to the amine (-NH ₂ ) group, which consists of one nitrogen atom and two hydrogen atoms.

The term "halogen" as used in chemical formulas includes atoms such as fluorine, bromine, chlorine, and iodine.

The term "ketone" as used in chemical formulas refers to a functional group consisting of one carbonyl group bonded to two hydrocarbon groups.

The term "cyano" as used in chemical formulas refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

The term "alkyl" as used in chemical formulas refers to a fully saturated branched or unbranched (or straight-chain or linear) hydrocarbon group.

Non-limiting examples of the above “alkyl” include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, iso-amyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, and the like.

One or more hydrogen atoms of the above "alkyl" may be substituted with a halogen, a C1-C20 alkyl group substituted with a halogen (e.g., CCF ₃ , CHCF ₂ , CH ₂ F, CCl ₃ , etc.), a C1-C20 alkoxy group, a C2-C20 alkoxyalkyl group, a hydroxy group, a nitro group, a cyano group, an amino group, or a C1-C20 alkyl group, a C2-C20 alkenyl group, a C2-C20 alkynyl group, a C1-C20 heteroalkyl group, a C6-C20 aryl group, a C6-C20 arylalkyl group, a C6-C20 heteroaryl group, a C7-C20 heteroarylalkyl group, a C6-C20 heteroaryloxy group, or a C6-C20 heteroaryloxyalkyl group.

The term "halogen-substituted C1-C20 alkyl group" refers to a C1-C20 alkyl group substituted with one or more halo groups, including, but not limited to, polyhaloalkyl containing monohaloalkyl, dihaloalkyl or perhaloalkyl. Monohaloalkyl is when the alkyl group has one iodine, bromine, chlorine or fluorine, and dihaloalkyl and polyhaloalkyl refer to alkyl groups having two or more identical or different halo atoms.

The term "alkoxy" used in chemical formulas represents alkyl-O-, wherein the alkyl is as described above. Non-limiting examples of the alkoxy include methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy, cyclopropoxy, cyclohexyloxy, etc. At least one hydrogen atom in the alkoxy group may be replaced with the same substituent as in the case of the alkyl group described above.

The term "alkoxyalkyl" as used in chemical formulas refers to an alkyl group substituted with the alkoxy group described above. One or more hydrogen atoms in the alkoxyalkyl group may be replaced with the same substituents as those in the alkyl group described above. Thus, the term "alkoxyalkyl" includes substituted alkoxyalkyl moieties.

The term "alkenyl" as used in chemical formulas refers to a branched or unbranched hydrocarbon having at least one carbon-carbon double bond. Non-limiting examples of alkenyls include vinyl, allyl, butenyl, isopropenyl, isobutenyl, etc., and one or more hydrogen atoms in the alkenyls may be replaced with the same substituents as in the alkyl groups described above.

The term "alkynyl" as used in chemical formulas refers to a branched or unbranched hydrocarbon group having at least one carbon-carbon triple bond. Non-limiting examples of such alkynyl groups include ethynyl, butynyl, isobutynyl, isopropynyl, and the like. One or more hydrogen atoms in the alkynyl group may be replaced with the same substituents as those in the alkyl group described above.

The term "alkylene oxide" used in chemical formulas refers to alkylene-O. Non-limiting examples of the alkylene include methylene, ethylene, propylene, butylene, etc. In addition, one or more hydrogen atoms in the "alkylene" may be substituted with a substituent similar to the alkyl group described above.

The term "cycloalkyl" as used in chemical formulas refers to an alkyl group forming a ring. The alkyl group is as described above. Non-limiting examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. One or more hydrogen atoms in the cycloalkyl group may be replaced with the same substituents as in the alkyl group described above.

The term "aryl" used in chemical formulas, alone or in combination, refers to an aromatic hydrocarbon containing one or more rings. The term "aryl" also includes groups in which an aromatic ring is fused to one or more cycloalkyl rings. Non-limiting examples of the "aryl" include phenyl, naphthyl, tetrahydronaphthyl, and the like. In addition, one or more hydrogen atoms in the "aryl" may be replaced with a substituent similar to the alkyl group described above.

The term "arylalkyl" means alkyl substituted with aryl. Examples of arylalkyl include benzyl or phenyl-CH ₂ CH ₂ -.

The term "aryloxy" used in chemical formulas means O-aryl, and examples of aryloxy include phenoxy, etc. One or more hydrogen atoms in the "aryloxy" may be replaced with a substituent similar to the alkyl group described above.

The term "heteroaryl" as used in chemical formulas refers to a monocyclic or bicyclic organic compound containing one or more heteroatoms selected from N, O, P, or S, with the remaining ring atoms being carbon. The heteroaryl group may contain, for example, 1 to 5 heteroatoms and 5 to 10 ring members. The S or N may be oxidized to have various oxidation states.

One or more hydrogen atoms in the above “heteroaryl” can be substituted with a substituent similar to that in the case of the above-described alkyl group.

The term "heteroarylalkyl" means alkyl substituted with heteroaryl.

The term "heteroaryloxy" refers to an -O-heteroaryl moiety. One or more hydrogen atoms in the heteroaryloxy may be replaced with a substituent similar to that for the alkyl group described above.

The term "heteroaryloxy" means alkyl substituted with heteroaryl. One or more hydrogen atoms in the heteroaryloxyalkyl may be replaced with a substituent similar to that in the case of the alkyl group described above.

The term "amino group" refers to a nitrogen atom covalently bonded to at least one carbon or heteroatom. The amino group includes, for example, -NH ₂ and substituted moieties. The term "amino group" includes alkylamino groups in which the nitrogen is bonded to at least one additional alkyl group, arylamino groups in which the nitrogen is bonded to at least one or more independently selected aryl groups, and diarylamino groups.

The term "substitution" in the above "substituted or unsubstituted" refers to a case where one or more hydrogen atoms in an organic compound are replaced with another atomic group to form a derivative, and the substituent refers to the introduced atomic group. The substituent may be a halogen atom, a hydroxy group, a nitro group, a cyano group, an amino group, an amidino group, an acetamino group, a hydrazine, a hydrazone, a carboxyl group, a sulfonyl group, a sulfamoyl group, a sulfonic acid group, a phosphoric acid, _{a C 1 to C 5 alkyl} group, a C ₁ to C ₅ alkoxy group, a C ₂ to C ₅ alkenyl group, a C ₂ to C ₅ alkynyl group, a C ₃ to C ₁₀ cycloalkyl group, a C ₆ to C ₁₀ aryl group, a C ₆ to C 10 heteroaryl group, a C 6 to C ₂₀ arylalkyl group, a C ₆ to C ₂₀ heteroarylalkyl group, or a combination thereof.

The term "isomer" in "stereoisomer" used herein refers to a compound that has the same molecular formula but has different connection methods or spatial arrangements of the constituent atoms within the compound. Isomers include, for example, structural isomers and stereoisomers. The stereoisomers may be diastereomers or enantiomers. Enantiomers are isomers that do not overlap with their mirror images, like the relationship between left and right hands, and are also called optical isomers. Enantiomers are distinguished as R (Rectus: clockwise) and S (Sinister: counterclockwise) when four or more substituents are different at the chiral center carbon. Diastereoisomers are stereoisomers that are not mirror images, and can be divided into cis-trans isomers due to the different spatial arrangement of atoms.

The term "solvate" as used herein refers to a compound solvated in an organic or inorganic solvent. The solvate is, for example, a hydrate.

The term "salt" as used herein refers to inorganic and organic acid addition salts of a compound. The pharmaceutically acceptable salt may be a salt that does not cause serious irritation to an organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. The inorganic acid salt may be a hydrochloride, a bromate, a phosphate, a sulfate, or a disulfate. The organic acid salt may be a formate, an acetate, a propionate, a lactate, an oxalate, a tartrate, a malate, a maleate, a citrate, a fumarate, a besylate, a camsylate, an edisyl salt, a trichloroacetic acid, a trifluoroacetate, a benzoate, a gluconate, a methanesulfonate, a glycolate, a succinate, a 4-toluenesulfonate, a galacturonate, an embronate, a glutamate, an ethanesulfonate, a benzenesulfonate, a p-toluenesulfonate, or an aspartate. The above metal salt may be a calcium salt, a sodium salt, a magnesium salt, a strontium salt, or a potassium salt.

The compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof may be isolated from Streptomyces sp., and specifically, may be isolated from Streptomyces sp. GC2 strain (Accession No.: KCTC15723BP), a lysate thereof, a culture medium thereof, and/or an extract of the culture medium.

Another aspect is to provide a Streptomyces sp. GC5 strain (Accession No.: KCTC15723BP) that produces a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof. The same parts as described above also apply to the above strain.

The above strain can produce thioamide compounds, stereoisomers, solvates, or pharmaceutically acceptable salts according to one aspect.

The above strain includes its variants. The variants may be, for example, those resulting from natural mutations or artificial mutations. Artificial mutations may be caused by physical mutagens, such as ultraviolet light, or chemical mutagens, such as basic compounds.

The above strain includes spores, mycelia, or cultures of the strain.

Another aspect provides a method for preparing a compound of formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, comprising the steps of: 1) culturing a strain of Streptomyces sp. GC5 (Accession No.: KCTC15723BP); and 2) isolating the compound of formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof from the strain, a lysate thereof, a culture medium thereof, or an extract of the culture medium. The same parts as described above also apply to the method.

In one embodiment, a method may be used, including a step of culturing a strain of Streptomyces sp. GC2 (Accession No.: KCTC15723BP) to produce a compound represented by the chemical formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof; and a step of isolating the compound represented by the chemical formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof from the strain, a lysate thereof, a culture medium thereof, and/or an extract of the culture medium.

The above method comprises a step of culturing a Streptomyces sp. GC2 strain (Accession No.: KCTC15723BP). The culturing step may be culturing the strain in a liquid medium or a solid medium. The medium may include, as a carbon source, for example, glucose, corn syrup, dextrin, starch, molasses, animal oil, or vegetable oil. The medium may include, as a nitrogen source, for example, wheat bran, soybeans, soybean meal, wheat, malt, cottonseed meal, fish meal, corn syrup, meat juice, yeast extract, malt extract, ammonium sulfate, sodium nitrate, or urea.

In one embodiment, the culturing step may be culturing using YEME medium and/or modified K medium, and specifically, may include a step of culturing Streptomyces sp. GC2 strain (accession number: KCTC15723BP) using YEME medium, and then culturing the cultured strain using modified K medium.

The above culturing may be performed under aerobic conditions with shaking or standing. The culturing temperature may be, for example, about 20°C to about 40°C, about 25°C to about 37°C, about 28°C to about 35°C, or about 30°C. The culturing time may be, for example, about 1 day to about 4 months, about 1 day to about 2 months, about 1 day to about 6 weeks, about 1 day to about 1 month, about 1 day to about 2 weeks, or about 1 day to about 1 week.

The step of separating may include a step of concentrating, centrifuging, filtering, or performing chromatography on the strain, its lysate, its culture medium, and/or the extract of the culture medium. The chromatography may be, for example, column chromatography, planar chromatography, paper chromatography, or thin-layer chromatography, depending on the form of the stationary phase. The chromatography may be, for example, gas chromatography, liquid chromatography, or affinity chromatography, depending on the physical properties of the mobile phase. The liquid chromatography may be, for example, high-performance liquid chromatography (HPLC). The chromatography may be, for example, ion exchange chromatography, size-exclusion chromatography, depending on the separation method. The chromatography may be, for example, normal phase chromatography or reverse phase chromatography.

Another aspect provides a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof. The same parts as described above also apply to the composition.

The term “treatment” in this specification means any action that improves or beneficially changes the symptoms of a cancer disease by administering the composition of the present invention.

The term “prevention” in this specification means any action by which the possibility of developing a cancer disease or disease is suppressed or delayed by administration of the composition of the present invention.

The term "cancer" as used herein refers to a tumor or a disease that forms a tumor due to the uncontrolled, excessive growth of body tissue. Such cancer may be solid or non-solid. Solid cancer refers to cancerous tumors that develop in organs such as the liver, lungs, breasts, and skin. Non-solid cancer refers to cancer that develops in the blood, also known as hematologic cancer.

The cancer may be gastric cancer, liver cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, colon cancer, bone cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, colon cancer, anal cancer, colon cancer, breast cancer, cervical cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma, and specifically, the cancer may be lung cancer, colon cancer, This may include one or more of stomach cancer, liver cancer, breast cancer, and pancreatic cancer.

The above cancer may be a cancer resistant to anticancer drugs.

The term "anticancer agent (or anticancer drug)" as used herein refers to a chemical substance used for treatment, including shrinkage, inhibition, and elimination of malignant tumors. The anticancer agent may be, for example, alkylating agents, antimetabolites, natural substances, hormones and their antagonists, targeted therapeutic agents, or a combination thereof. The anticancer agent may include, for example, fluorouracil (5-FU), irinotecan, etoposide, gemcitabine, and paclitaxel.

The term "drug resistance" as used herein refers to a condition in which cancer symptoms do not improve, alleviate, alleviate, or respond to anticancer drug treatment due to extremely low sensitivity to the treatment. Anticancer drug resistance can occur when a cancer develops resistance to a specific anticancer drug from the outset, or when the cancer cells, previously unresistant, change in their properties after prolonged treatment, rendering them insensitive to the same drug.

The pharmaceutical composition may further comprise a known active ingredient having anticancer activity. The known active ingredient having anticancer activity may be an anticancer agent. The anticancer agent may be 5-fluorouracil, irinotecan, etoposide, gemcitabine, paclitaxel, oxaliplatin, leucovorin, capecitabine, or a combination thereof. The compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, and the anticancer agent may be a single or separate composition for simultaneous or sequential administration.

The compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof has a cancer cell growth inhibitory effect such as inducing apoptosis of cancer cells and/or inhibiting cell proliferation of cancer cells, thereby effectively treating/improving or preventing cancer diseases.

In addition, the compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof can improve and/or enhance the effect of cancer treatment by reducing the resistance of cancer or cancer cells to anticancer drugs or increasing the sensitivity/sensitivity to anticancer drugs.

The pharmaceutical composition may include a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" may refer to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity or properties of the injected compound. Here, "pharmaceutically acceptable" means that the carrier does not inhibit the activity of the active ingredient and does not exhibit toxicity exceeding the adaptability of the subject of application (prescription). Any carrier commonly used in the art and pharmaceutically acceptable for the pharmaceutical composition may be used. Non-limiting examples of the above carriers include lactose, dextrose, maltodextrin, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, glycerol, ethanol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. These may be used alone or in combination of two or more. The pharmaceutical composition may be prepared as an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The above pharmaceutical composition can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions, each according to a conventional method.

When formulating the above pharmaceutical composition, it may be prepared using a diluent or excipient such as a generally used filler, bulking agent, binder, wetting agent, disintegrant, or surfactant, but may not be limited thereto.

When the above pharmaceutical composition is manufactured into an oral dosage form, it can be manufactured into a dosage form such as powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, etc., using a suitable carrier according to a method known in the art. At this time, examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, etc.; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyols, and vegetable oils. In case of formulation, the formulation may include diluents and/or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants, as needed.

When the above pharmaceutical composition is prepared as a parenteral dosage form, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories using a suitable carrier according to a method known in the art. When formulated as an injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, and preferably, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, sterile water for injection, and isotonic solutions such as 5% dextrose can be used. When formulated as a transdermal dosage form, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pastes, liniments, aerosols, etc. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide, and in the case of formulating it as a suppository, the base can be witepsol, tween 61, polyethylene glycol, cacao butter, laurin butter, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, sorbitan fatty acid ester, etc.

The pharmaceutical composition may be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" refers to an amount sufficient to treat or prevent a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention, and the effective dosage level may be determined based on factors including the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, drugs used in combination or simultaneously with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a component known to exhibit a therapeutic effect on a known cancer disease. It is important to take all of the above factors into consideration and administer an amount that can achieve the maximum effect with the minimum amount without side effects.

The dosage of the pharmaceutical composition may be determined by a person skilled in the art in consideration of the intended use, the degree of toxicity of the disease, the patient's age, weight, sex, medical history, or the type of substance used as the active ingredient. For example, the pharmaceutical composition of the present invention may be administered at about 0.1 ng to about 1,000 mg/kg, preferably 1 ng to about 100 mg/kg, per adult, and the frequency of administration of the composition of the present disclosure is not particularly limited thereto, but may be administered once a day or administered in divided doses several times. The dosage or frequency of administration does not limit the scope of the present disclosure in any way.

Another aspect provides a method for treating or preventing cancer, comprising administering to a subject a pharmaceutical composition for treating or preventing cancer. The same principles as described above apply to the method.

The term "subject" as used herein may include, without limitation, mammals, birds, reptiles, farmed fish, etc., including dogs, cats, rats, livestock, humans, etc., that are suffering from or at risk of suffering from cancer, and the subject may exclude humans.

The pharmaceutical composition described above may be administered in single or multiple doses in a pharmaceutically effective amount. The composition may be formulated and administered in the form of a liquid, powder, aerosol, injection, infusion solution (Ringel), capsule, pill, tablet, suppository, or patch. The pharmaceutical composition for cancer prevention or treatment may be administered via any conventional route, as long as it can reach the target tissue.

The pharmaceutical composition may be administered, but is not particularly limited thereto, via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and rectal administration, depending on the intended purpose. However, when administered orally, it may be administered in an unformulated form, and since the active ingredient of the pharmaceutical composition may be denatured or decomposed by gastric acid, the oral composition may be administered orally in a form that coats the active agent or is formulated to protect it from decomposition in the stomach, or in the form of an oral patch. In addition, the composition may be administered by any device that allows the active ingredient to move to the target cell.

Another aspect provides a food composition for preventing or improving cancer, comprising a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof. The same parts as described above also apply to the composition.

The term “improvement” in this specification means any act of improving or beneficially altering the symptoms of a cancer disease by administration of the composition of the present invention.

The term "food" in this specification includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

The above food can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. In addition, the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition can be manufactured in various forms of formulation, and unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material, and is highly portable, so the food composition of the present invention can be taken as a supplement to enhance the effect of improving cancer.

The above food composition may be a health functional food composition.

The term "health functional food" in this specification refers to a food manufactured and processed using raw materials or ingredients with functionality useful to the human body as defined in Act No. 6727 on Health Functional Foods, and "functionality" refers to obtaining a useful effect for health purposes, such as regulating nutrients or physiological effects for the structure and function of the human body.

The above health food refers to a food that has a more active health maintenance or promotion effect than general food, and health supplement food refers to a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement may be used interchangeably. Specifically, the above health functional food refers to a food that is made by adding a composition to food materials such as beverages, teas, spices, gum, and confectionery, or by manufacturing it in the form of capsules, powder, suspension, etc., and means that when consumed, it brings about a specific health effect. However, unlike general drugs, it has the advantage of not having the side effects that can occur with long-term use of drugs because it is made from food.

The above food composition can be used very usefully because it can be consumed on a daily basis and is expected to be highly effective in improving cancer.

The food composition may further comprise a physiologically acceptable carrier, and the type of the carrier is not particularly limited, and any carrier commonly used in the art may be used. Specifically, the food composition may comprise additional ingredients commonly used in food compositions to improve odor, taste, sight, etc. For example, the food composition may comprise vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, the food composition may comprise minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). In addition, the food composition may comprise amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), bactericides (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar colorants, etc.), color developers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (glucose fillers), film-forming agents, gum-forming agents, foam suppressants, solvents, and improvers. The above additives can be selected according to the type of food and used in an appropriate amount.

The above food composition can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient mixed can be appropriately determined depending on its purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the food composition of the present invention can be added to the food or beverage in an amount of 50 parts by weight or less, specifically 20 parts by weight or less. However, when consumed for a long period of time for health and hygiene purposes, the content below the above range can be included, and since there is no problem in terms of safety, the active ingredient can also be used in an amount above the above range.

As an example of the food composition, the above-mentioned food composition may be used as a health beverage composition, and in this case, various flavoring agents or natural carbohydrates may be contained as additional ingredients, as in a conventional beverage. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin and stevia extract; or a synthetic sweetener such as saccharin and aspartame. The proportion of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g, per 100 mL of the health beverage composition of the present invention.

In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination. The proportion of these additives is not particularly important, but is typically selected within the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

The above food composition may contain the compound of the present invention in various weight % if it can exhibit an effect of improving or preventing cancer, and specifically may contain 0.00001 to 100 weight % or 0.01 to 80 weight % relative to the total weight of the food composition, but is not limited thereto.

Another aspect provides the use of a compound represented by the above chemical formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating cancer. The same portions described above also apply to the above use.

Another aspect provides a use of a compound represented by the above formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, for use in preventing or treating cancer. The same portions described above also apply to the above use.

The novel thioamide compound of the present invention, its stereoisomer, solvate, or pharmaceutically acceptable salt not only has anticancer efficacy against various types of cancer, but also has excellent anticancer activity against anticancer drug-resistant cancer, and thus can be used to prevent or treat various types of cancer.

Figure 1 is a drawing showing a photograph of a medium in which Streptomyces sp. GC2 strain (accession number: KCTC15723BP) spread on an agar plate was cultured.

Figure 2 is a diagram showing the cell proliferation inhibitory effect on the PANC-GR cell line according to the combination of thioguanamide B and gemcitabine.

Figure 3 is a diagram showing the tumor inhibition efficacy in a PANC-GR cell line tumor xenograft animal model by combined administration of thioguanamide B and gemcitabine.

Figure 4 is a diagram showing changes in body weight according to combined administration of thioguanamide B and gemcitabine.

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

Example 1: Isolation and structural confirmation of thiogochangamide

1-1: Isolation of thioguanamide-producing strain

Streptomyces sp. GC2, a strain producing thioguanamide A and B, was isolated from a soil sample collected from the Gochang wetland, South Korea, in May 2020. The collected soil sample was diluted in sterile water, and various isolation agar media were used for strain isolation. The GC2 strain was isolated from Czapek-Dox medium cultured at 30°C for 2 weeks (Fig. 1). 16S rRNA gene sequence analysis confirmed that the isolated GC2 strain belonged to the genus Streptomyces. The strain was deposited with the Korea Research Institute of Bioscience and Biotechnology on December 6, 2023 (Accession Number: KCTC15723BP).

1-2: Cultivation of Streptomyces sp. GC2 strain

The Streptomyces GC2 strain isolated in Example 1-1 above was cultured using the following method.

Step 1: Inoculate GC2 strain onto YEME solid agar medium (10 g malt extract, 4 g yeast extract, 4 g glucose, 18 g agar per liter of water) and culture at 30°C for one week.

Step 2 Cultivation: Prepare 50 mL of YEME liquid medium (10 g of malt extract, 4 g of yeast extract, 4 g of glucose per liter of water) in a 125 mL flask, inoculate with spores of the GC2 strain grown on the solid medium, and culture in an incubator at 30 °C and 200 rpm for 2 days.

Step 3 culture: Prepare 200 mL of modified K liquid medium (4 g yeast extract, 5 g malt extract, 5 g soybean, 5 g starch, 5 g mannitol, 2 g glucose, 6 g glycerol per liter of water) in a 500 mL flask, inoculate with 5 mL of the step 2 culture, and culture in an incubator at 30 °C and 170 rpm for 3 days.

Step 4 Cultivation: Prepare 1 L of modified K liquid medium (4 g yeast extract, 5 g malt extract, 5 g soybean, 5 g starch, 5 g mannitol, 2 g glucose, 6 g glycerol per 1 L of water) in a 2.5 L yield-enhancing flask, inoculate with 15 mL of the Step 3 culture, and culture in an incubator at 30 °C and 170 rpm for 6 days.

1-3: Isolation and purification of thioguanamide

In order to isolate and purify thioguanamide A and B from the Streptomyces sp. GC2 strain cultured in Example 1-2, the following experiments were performed.

First, a culture of the Streptomyces sp. GC2 strain cultivated in the above Example 1-2 was obtained, and a total of 200 L of the culture solution was extracted twice with 300 L of ethyl acetate (EtOAc) using a separatory funnel. The water remaining in the EtOAc layer was completely removed with anhydrous sodium sulfate and concentrated in vacuo using a reduced pressure evaporator to obtain 50 g of a dry extract. 10 g of the extract was dissolved in methanol (MeOH) and then adsorbed onto 5 g of Celite. The adsorbate was loaded onto a C18 reversed-phase open column (34 x 150 mm), and 20%, 40%, 60%, 80%, and 100% MeOH solutions (500 mL each) were applied to generate five fractions (repeated five times since a total of 50 g of extract was collected). Analysis of 10 μL aliquots of each fraction by LC/MS confirmed that thioguanamides A and B were detected in the 80% and 100% MeOH-H ₂ O fractions, respectively.

Therefore, 80% and 100% MeOH-H ₂ O fractions were evaporated and redissolved in MeOH and subjected to reversed-phase chromatography. The conditions for the first-step reversed-phase chromatography were as follows: (Column used: Kromasil C18, 5 μm, 250 × 10 mm, Flow rate: 2 mL/min, Detection UV: 270 nm, Solvent system of 30% to 60% CH 3 CN-H 2 O containing 0.1 formic acid over 40 min). Thioguanamides A and B eluted at 29 and 31 min, respectively. These were further purified in the second-step chromatography under the following conditions: (Column used: IB CHIRALPAK, 5 μm, 250 × 4.6 mm, 40% CH 3 CN-H 2 O, Detection UV: 270 nm, Flow rate: 0.7 mL/min). Thiochochanamide A and B were eluted at 30 and 33 minutes, respectively.

1-4: Confirmation of the chemical structure of thioguanamide

The structures of thioguanamides A and B isolated and purified in Examples 1-3 were confirmed based on 1D and 2D nuclear magnetic resonance (NMR) spectra. Nuclear magnetic resonance spectra ( ¹ H NMR, ¹³ C NMR) were obtained using 800 MHz or 200 MHz NMR, and Acetonitrile -d ₃ was used.

As a result of the above analysis, the chemical structures of thioguanamide A and B were confirmed as follows.

(1) Thiogazanamide A (compound of chemical formula 2)

1) Molecular formula: C ₅₆ H ₈₇ N ₁₄ O ₉ S ₆ ⁺

2) Molecular quantity: 1291

3) Colors: colorless

4) ¹ H-NMR (Acetonitrile- d ₃ , 800 MHz): listed in Table 1

5) ¹³ C-NMR (Acetonitrile- d ₃ , 200 MHz): listed in Table 1

[Chemical Formula 2]

(2) Thiogazanamide B (compound of chemical formula 3)

1) Molecular formula: C ₅₆ H ₈₇ N ₁₄ O ₁₀ S ₆ ⁺

2) Molecular quantity: 1307

3) Colors: colorless

4) ¹ H-NMR (Acetonitrile-d ₃ , 800 MHz): listed in Table 1

5) ¹³ C-NMR (Acetonitrile-d ₃ , 200 MHz): listed in Table 1

[Chemical Formula 3]

Position Thiogochangamide A (1) Thiogochangamide B (2) ¹ H NMR ¹³ C NMR ¹⁵ N NMR ¹ H NMR ¹³ C NMR δ _H , mult(J in Hz) δ _C δ _N δ _H , mult
(J in Hz) δ _C Residue 1 Lactic acid Lactic acid 1 1.26, d (7.0) 20.8 1.27 d (7.0) 20.9 2 4.21, m 69.3 4.22 q (7.0) 69.3 3 180.2 180.0 Residue 2 valine valine 4-NH 7.60, d (5.0) 110.8 7.60, overlap 4 4.46, m 69.3 4.46, dd (5.0) 69.3 5 2.48, m 32.2 2.47, m 32.2 6 0.97, d (7.0) 17.6 0.97, d(7.0) 17.7 7 1.06, d (7.0) 20.2 1.06, d(7.0) 20.2 8 206.8 206.7 Residue 3 methionine methionine 9-NH 10.60, br s 164.5 10.52, br s 9 5.75, m 67.8 5.73, m 67.7 10 2.24, m
2.57, m 31.9 2.24, m
2.52, m 31.9 11 2.53, m
2.68, m 31.4 2.53, m
2.67, m 31.3 12 2.07, s 15.2 2.07, s 15.2 13 206.6 206.5 Residue 4 alanine alanine 14-NH Not detected 170.2 10.87, br s 14 4.60, q (7.0) 58.5 4.60, q (7.0) 58.4 15 1.51, d (7.0) 16.6 1.50, d (7.0) 16.6 16 176.4 176.3 Residue 5 alanine alanine 17-NH 8.25 br s 120.7 8.22 br s 17 4.52 qd (7.0, 3.0) 61.6 4.52 qd (7.0, 3.0) 61.4 18 1.58, d (7.0) 19.4 1.57, d (7.0) 19.5 19 209.3 209.1 Residue 6 alanine alanine 20-NH 8.87 d (6.0) 158.9 8.87, d (6.0) 20 5.32 m 65.1 5.33, m 65.0 21 1.81, d (7.0) 19.4 1.79, d (7.0) 19.5 22 206.5 206.5 Residue 7 alanine alanine 23-NH 8.80 d (6.0) 162.4 8.87, d (6.0) 23 4.74, m 58.9 4.73, m 58.8 24 1.78, d (7.0) 17.1 1.75, d (7.0) 17.1 25 174.0 174.0 Residue 8 27-methyl- S -30-aminovinyl-cysteine 27-methyl- S -30-aminovinyl-cysteine 26-NH 7.86, d (7.0) 108.3 7.84, d (7.0) 26 4.00, m 58.1 4.04, m 58.0 27 3.08, q (7.0) 49.5 3.10, q (7.0) 49.8 28 1.38, d (7.0) 22.4 1.38, d (7.0) 22.4 29 5.55, d (7.0) 100.3 5.62, d (7.0) 101.8 30 7.29,dd (11.0, 7.0) 134.7 7.32,
dd (11.0, 7.0) 134.0 30-NH 10.47, d (11.0) 10.60, d (11.0) 31 172.0 172.1 Residue 9 valine valine 32-NH 6.64, d (6.0) 118.6 6.69, d (6.0) 32 3.75, m 64.4 3.81, m 64.3 33 2.04, m 31.1 2.05, m 31.1 34 0.96, d (7.0) 19.6 0.98, d (7.0) 19.6 35 1.08, d (7.0) 20.7 1.09, d (7.0) 20.6 36 174.1 174.3 Residue 10 alanine alanine 37-NH 7.19, overlap 121.3 7.19, overlap 37 3.84, m 53.4 3.80, m 53.4 38 1.10, d (7.0) 17.1 1.09, d (7.0) 17.0 39 174.8 174.6 Residue 11 phenylalanine phenylalanine 40-NH 7.84, d (8.5) 107.0 7.80, d (8.5) 40 4.48, m 54.9 4.37, m 54.8 41 2.85,dd (13.0, 13.0)
3.19,
dd (13.0, 3.0) 37.3 2.82,
dd (13.0, 13.0)
3.15,
dd (13.0, 3.0) 37.1 42 139.4 139.2 43, 47 7.31, d (7.0) 130.2 7.27, overlap 130.2 44, 46 7.29, dd (7.0, 7.0) 129.0 7.25, overlap 129.1 45 7.18, overlap 127.4 7.17, overlap 127.4 48 172.9 172.9 Residue 12 N,N-dimethylhistidinium β-hydroxy- N , N -dimethylhistidinium 49-NH 7.63, d (7.0) 112.9 7.61 overlap 49 4.22, m 54.2 4.15,dd (8.0, 8.0) 58.0 50 3.29, ddd(16.0, 4.0, 0.5)
3.46, ddd
(16.0, 11.0, 0.5) 22.9 5.77, d (8.0) 63.6 51 134.0 134.3 52 3.75, s 34.4 3.91, s 35.4 53 8.28, s 136.7 8.31, s 137.9 53-N 171.9 54 3.81, s 36.6 3.83, s 36.7 55 7.48, br s 122.7 7.60, br s 123.7 55-N 169.6 56 168.0 169.0

Example 2: Confirmation of anticancer activity of thioguanamide

To confirm the anticancer activity of thioguanamide A and B obtained in Example 1, the following experiments were performed.

First, to evaluate cytotoxicity against various cancer cell lines, human lung cancer (A549), colon cancer (HCT116), gastric cancer (SNU638), liver cancer (SK-HEP-1), and breast cancer (MDA-MB-231) cells were provided by the Korea Cell Line Bank. The cancer cell lines were cultured in a medium (RPMI-1640 medium for A549, HCT116, and SNU638 cells, Dulbecco's Modified Eagle's medium (DMEM) for MDA-MB-231 and SK-HEP-1 cells) containing 10% heat-inactivated fetal bovine serum (FBS) and antibiotic-antimycotic solution (100 units/mL penicillin G sodium, 100 μg/mL streptomycin, and 250 ng/mL amphotericin B).

To evaluate the cytotoxicity of thioguanamides A and B against cancer cells, cell proliferation was determined using the sulforhodamine B (SRB) assay. Specifically, the cancer cells were seeded in 96-well plates and cultured for 30 minutes (day 0 control) or treated with thioguanamides A and B and etoposide for the indicated times. Etoposide served as a positive control. After culturing for 72 hours, the cells were fixed, dried, and stained with 1% acetic acid containing 0.4% SRB. Unbound dye was then washed away, and the stained cells were suspended in 10 mM Tris (pH 10.0). The absorbance was measured at 515 nm, and cell proliferation was determined using the following mathematical formula:

Cell proliferation(%)= (average absorbance _sample - average absorbance _zeroday ) / (average absorbance _control - average absorbance _zeroday ) x 100

IC ₅₀ values were calculated by nonlinear regression analysis using TableCurve 2D v5.01 software (Systant Software Inc., Richmond, CA, USA).

As a result, thioguanamides A and B showed relatively high inhibitory activity against all five cancer cell lines, and in particular, it was confirmed that they showed superior anticancer efficacy than the positive control, etoposide, in colon cancer and breast cancer cell lines (Table 2).

IC ₅₀ (μM) SNU638 SK-Hep-1 A549 HCT116 MDA-MB-231 Thiogochangamide A <0.4 1.53 <0.4 <0.4 2.37 Thiogochangamide B <0.4 1.34 <0.4 <0.4 1.78 Etoposide 0.17 0.46 0.19 0.9 4.84

Example 3: Confirmation of anticancer activity of thioguanamide against resistant cancer cells.

In order to confirm the anticancer activity of thioguanamide obtained in Example 1 above against resistant cancer, an anticancer drug resistance model was constructed and its anticancer efficacy was evaluated.

First, the PANC-1 pancreatic cancer cell line was purchased from the Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM medium supplemented with 10% heat-inactivated FBS (Gibco, Grand Island, NY), 100 units/mL penicillin, and 100 μg/mL streptomycin. The gemcitabine-resistant cell line, PANC-GR, was established by culturing PANC-1 cells with increasing doses of gemcitabine (0.1–2 μM). Cells were passaged 2–3 times per week in a 37°C incubator under 5% _CO2 conditions. Gemcitabine used was purchased from Sigma-Aldrich (St. Louis, MO, USA).

The anticancer efficacy of thioguanamide B and gemcitabine was evaluated against the PANC-1 pancreatic cancer cell line and the gemcitabine-resistant cell line PANC-GR, respectively. Thioguanamide B exhibited anticancer efficacy against PANC-1 (IC ₅₀ : 1.23 μM) and PANC-GR (IC ₅₀ : 0.48 μM) cells, whereas gemcitabine exhibited anticancer efficacy only against PANC-1 cells (IC ₅₀ : 2.1 μM) and did not exhibit anticancer effect against the resistant cell line PANC-GR cells (IC ₅₀ : > 50 μM) (Table 3). Based on the above results, it can be seen that thioguanamide B can overcome gemcitabine resistance acquired in pancreatic cancer.

IC ₅₀ (μM) PANC-1 PANC-GR thiogochangamideB 1.23 0.48 Gemcitabine 2.1 >50

Next, to confirm the drug combination effect of thioguanamide B and gemcitabine, PANC-GR cells were treated with the combination of thioguanamide B and gemcitabine for 72 hours, and then cell viability was confirmed using the SRB assay, and the combination index (CI) was calculated using the Chou-Talalay method using the following mathematical formula:

CI = D ₁ /(D _x ) ₁ + D ₂ /(D _x ) ₂

(D ₁ and D ₂ are the concentrations of the combined test compounds that achieve a synergistic effect, and (D _x ) ₁ and (D _x ) ₂ are the single-administration concentrations.)

The above results confirmed that thioguanamide B restored sensitivity to gemcitabine in combination with gemcitabine in the gemcitabine-resistant PANC-1 cell line, and the drug combination showed a synergistic effect on anticancer efficacy as CI < 1 (Table 4 and Fig. 2).

Dose (μM) Combination
Index (CI) Description ThiogochangamideB Gemcitabine 0.25 5 0.25 Strong synergism 0.5 10 0.28 Strong synergism 1 20 0.52 synergism

Next, the PANC-GR cell line contracted above was subcutaneously injected into BALB/c-nude mice (6-week-old, male) at a cell concentration of ^1x107 /mL. When the tumor size reached 100 ^mm3 , thioguanamide B (1 mg/kg) and gemcitabine (20 mg/kg) were administered intraperitoneally three times a week at a fixed concentration, and a combination of gemcitabine and thioguanamide B (gemcitabine 20 mg/kg; thioguanamide B 1 mg/kg) was co-administered three times a week. The change in tumor size and body weight were measured twice a week. After tumor transplantation, the tumor size was measured using a caliper, and the tumor volume was calculated using the following mathematical formula:

Tumor volume (mm ³ ) = (width) x (length) x (height) x π/6

In a tumor xenograft model of the PANC-GR cell line, a gemcitabine-resistant pancreatic cancer cell line, the antitumor activity of thioguanamide B was confirmed by single administration three times a week for 23 days and in combination with gemcitabine. As a result, the tumor size in the thioguanamide B group was reduced by 52.21% compared to the gemcitabine monotherapy group with low sensitivity, and in the combination therapy group, the tumor size was reduced by 65.09%, confirming excellent antitumor activity (Figs. 3 and 4).

The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[Accession number]

Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

Accession number: KCTC15723BP

Date of acceptance: 20231206

Claims (15)

A compound represented by the formula 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R ¹ to R ¹⁸ are each independently hydrogen, hydroxyl group, amino group, halogen group, ketone group, cyano group, -C(=O)R _a , -C(=O)OR _a , -OCO(OR _a ), -C=N(R _a ), -SR _a , -S(=O)R _a , -S(=O) ₂ R _a , -PR _a , A substituted or unsubstituted C ₁ to C ₂₀ alkyl group, a substituted or unsubstituted C ₁ to C ₂₀ alkoxy group, a substituted or unsubstituted C ₂ to C ₂₀ alkenyl group, a substituted or unsubstituted C ₂ to C ₂₀ alkynyl group, a C ₂ to C ₂₀ alkylene oxide group, a substituted or unsubstituted C ₃ to C ₃₀ cycloalkyl group, a substituted or unsubstituted C ₆ to C ₃₀ aryl group, a substituted or unsubstituted C ₆ to C ₃₀ aryloxy group, a substituted or unsubstituted C ₆ to C ₃₀ heteroaryl group, or a combination thereof, wherein R _a is hydrogen, a substituted or unsubstituted C ₁ -C ₁₀ alkyl group, or a substituted or unsubstituted C ₆ -C ₂₀ aryl group. A compound, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ is hydrogen or a hydroxy group in claim 1. A compound, ^{a stereoisomer} , a solvate, or a pharmaceutically acceptable salt thereof, in claim 1, wherein at least one of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R 8 , R ⁹ , R 10 , R ¹² and R ¹³ is a substituted or unsubstituted C ₁ to C ₁₀ alkyl group. A compound, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein at least one of ^{R 1} , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R 10 , R ¹² and R ¹³ is a methyl group. A compound, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein at least one of R ² and R ⁹ is an isopropyl group. A compound, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, wherein R ¹⁶ is hydrogen or a hydroxy group in claim 1. In claim 1, the compound represented by the chemical formula 1 is a compound represented by the chemical formula 2 or 3, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof: [Chemical Formula 2] [Chemical Formula 3] . In claim 1, the compound represented by the chemical formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof is isolated from Streptomyces sp. A strain of Streptomyces sp. GC5 (Accession No.: KCTC15723BP) producing a compound of claim 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof. 1) A step of culturing the Streptomyces sp. GC5 strain (Accession No.: KCTC15723BP); and 2) A step of isolating the compound of claim 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof from the strain, a lysate thereof, a culture medium thereof, or an extract of the culture medium. A method for preparing a compound of claim 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof, comprising: A pharmaceutical composition for preventing or treating cancer, comprising the compound of claim 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof. In claim 11, the cancer comprises at least one of stomach cancer, liver cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, colon cancer, bone cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, colon cancer, anal cancer, colon cancer, breast cancer, cervical cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. Composition. A pharmaceutical composition according to claim 11, wherein the cancer is a cancer resistant to an anticancer drug. A pharmaceutical composition according to claim 11, wherein the composition further comprises an anticancer agent. A food composition for preventing or improving cancer, comprising the compound of claim 1, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.