TW202412813A - Selenium-containing chalcone compound and preparation method thereof, pharmaceutical composition containing the same, and use for preparing pharmaceutical composition for preventing or treating colorectal cancer having better therapeutic effects on colorectal cancer than the chemotherapy drug 5-FU of the prior art - Google Patents

Abstract

The present invention relates to a selenium-containing chalcone compound and a preparation method thereof, a pharmaceutical composition containing the same, and a use for preparing a pharmaceutical composition for preventing or treating colorectal cancer. The selenium-containing chalcone compound is represented by formula (I) or pharmaceutically acceptable salts thereof. The present invention uses a simple synthesis method to synthesize four selenium-containing chalcone compounds, which have better therapeutic effects on colorectal cancer than the chemotherapy drug 5-FU of the prior art.

Description

Selenium-containing chalcone compound and preparation method thereof, pharmaceutical composition containing the same, and use of the pharmaceutical composition for preparing a pharmaceutical composition for preventing or treating colorectal cancer

The present invention relates to a selenium-containing chalcone compound, and in particular to the use of the selenium-containing chalcone compound in preparing a pharmaceutical composition for preventing or treating colorectal cancer.

According to statistics from the World Health Organization (WHO), there are as many as 1 million new cases of colorectal cancer and 700,000 deaths from colorectal cancer each year, making it the third most common cancer. Most colorectal cancers are caused by lifestyle and aging, while a small number are caused by genetic diseases. Risk factors include diet, obesity, smoking, and lack of exercise. Colorectal cancer has no obvious symptoms in the early stages and is not discovered until the middle or late stages of the disease. Treatment methods include surgery, radiotherapy, chemotherapy, targeted therapy, or a combination of the above. However, it is still difficult to completely cure colorectal cancer with existing drugs.

Selenium is an essential trace nutrient for the human body and is beneficial to human health. Selenium deficiency is closely related to cardiovascular disease, diabetes, thyroid dysfunction, etc. Selenocystein (SeCys) is a selenol group that replaces the thiol group in cysteine, that is, an amino acid that binds to selenoproteins. Selenoproteins may play an important role in preventing cancer. Among the 25 selenoproteins identified in mammals, glutathione peroxidases (GPx), iodothyronine deiodinase (ID), and thioredoxin reductase (TrxR) are related to cancer. Selenium-containing enzymes regulate many functions of mammals, including metastasis, cell apoptosis, tumor growth, blood sugar, antioxidants, and drug resistance.

Selenium-containing drugs can be used in anti-inflammatory, antioxidant, antiviral and anticancer activities, and are therefore often used as antioxidants, anti-inflammatory agents and anticancer agents. However, there is very little research on the effects of selenium-containing drugs on colorectal cancer, especially selenium-containing chalcone compounds, which have not been fully studied.

In view of this, the inventors of the present invention deeply understand the shortcomings and defects of the previous case and are eager to improve and innovate it. After years of research, they finally successfully developed a chalcone compound containing selenium. This compound has been proven to have anti-colorectal cancer effects and also provides a novel and safe treatment strategy for colorectal cancer treatment.

To achieve the above object, the present invention provides a selenium-containing chalcone compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof: In formula (I), n is an integer from 0 to 4, and each R is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ or -SR', wherein each R' is independently a substituted or unsubstituted alkyl group.

In one embodiment of the present invention, n is an integer from 0 to 2, and R is -OH.

In one embodiment of the present invention, formula (I) is represented by any one of the following compounds: , , , .

The present invention further provides a pharmaceutical composition comprising any of the above-mentioned selenium-containing chalcone compounds and a pharmaceutically acceptable carrier.

In one embodiment of the present invention, the pharmaceutical composition is a solution, suspension, emulsion, powder, tablet, pill, syrup, buccal tablet, tablet, chewing gum, slurry or capsule.

The present invention further provides a use of a selenium-containing chalcone compound for preparing a pharmaceutical composition for preventing or treating colorectal cancer, wherein the selenium-containing chalcone compound comprises any of the above-mentioned selenium-containing chalcone compounds.

In one embodiment of the present invention, the effective dose of the selenium-containing chalcone compound is 10 μM to 200 μM.

The present invention further provides a method for preparing a selenium-containing chalcone compound, comprising the steps of a chemical reaction represented by the following formula (II): In formula (II), n is an integer from 1 to 5, R ₁ is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ , -SR' or methoxy methyl (MOM); R ₂ is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ or -SR'; wherein R' is independently substituted or unsubstituted alkyl.

In one embodiment of the present invention, when any one of R ₁ is methoxymethyl ether, it further comprises the step of reacting the product of the reaction of formula (II) with hydrochloric acid at 100 ^° C. to convert the methoxymethyl ether into -OH.

The present invention provides a selenium-containing chalcone compound and a preparation method thereof. The technology is very easy to operate, and the raw chemical drugs are very cheap and have a moderate yield. The pure product can be obtained by chromatography purification or vacuum filtration. Compared with the known chemical drug 5-FU, the treatment effect of the compound on colorectal cancer is better.

[Definition of Terms] This specification widely uses many technical and scientific terms commonly used in the field of biotechnology. In the following description, in order to have a clear and consistent understanding of this specification and the scope of the patent application and the scope of these terms, the following definitions are provided. Other terms not specifically defined below have the meanings commonly understood by professionals in the relevant field.

Unless otherwise specified, "or", "and", and" in this invention all refer to "or/and". In addition, the terms "include" and "include" are not open conjunctions with any restrictions. The above paragraphs are only systematic references and should not be interpreted as limiting the subject matter of the invention.

All numerical values set forth in this invention may have a standard technical measurement error (standard deviation) of ± 10%. The term "about" is intended to mean ± 10%, ± 5%, ± 2.5%, or ± 1% relative to a given value, that is, "about" 20% means 20 ± 2%, 20 ± 1%, 20 ± 0.5%, or 20 ± 0.25%.

"Substituted" as used herein means that the hydrogen atom bonded to the carbon atom of the compound is changed to another substituent, and the substitution position is not limited as long as it is a position where the hydrogen atom is substituted, that is, a position where a substituent can be substituted, and when two or more substituents are substituted, the two or more substituents may be the same or different from each other;

The alkyl group described in the present invention includes a straight chain or branched chain having 1 to 60 carbon atoms, and may be further substituted with other substituents. The number of carbon atoms in the alkyl group may be 1 to 60, specifically 1 to 40, and more specifically 1 to 20. Specific examples thereof may include methyl, ethyl, propyl, n-propyl, isopropyl, butyl, n-butyl, isobutyl, tertiary butyl, di-butyl, 1-methyl-butyl, 1-ethyl-butyl, pentyl, n-pentyl, isopentyl, neopentyl, tertiary pentyl, hexyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 4-methyl-2-pentyl, 3,3-dimethylbutyl, 2-ethylbutyl, heptyl, n-heptyl, 1-methylhexyl, cyclopentylmethyl, cyclohexylmethyl, octyl, n-octyl, tertiary octyl, 1-methylheptyl, 2-ethylhexyl, 2-propylpentyl, n-nonyl, 2,2-dimethylheptyl, 1-ethyl-propyl, 1,1-dimethyl-propyl, isohexyl, 2-methylpentyl, 4-methylhexyl, 5-methylhexyl and the like, but are not limited thereto.

The term "pharmaceutically acceptable" as used herein means that the substance or composition must be compatible with other ingredients of its pharmaceutical formulation and does not aggravate the symptoms of the patient.

The "pharmaceutically acceptable salt" in the present invention refers to a salt prepared from a pharmaceutically non-toxic base or acid including an inorganic base or an organic base and an inorganic acid or an organic acid. The salt of the alkaline compound included in the term "pharmaceutically acceptable salt" refers to a non-toxic salt of the compound of the present invention which is usually prepared by reacting the free base with a suitable organic acid or an inorganic acid. Representative salts of the alkaline compounds of the present invention include, but are not limited to, the following salts: acetate, ascorbate, adipate, alginate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, camphorate, camphorsulfonate, dextrorotatory camphorsulfonate, carbonate, chloride, clavulanate, citrate, cyclopentanepropionate, diethylacetate, diisocyan ... gluconate, dihydrochloride, dodecyl sulfate, edetate, edisulphonate, estolate, ethanesulfonate, ethanesulfonate, formate, fumarate, gluceptate, glucoheptanoate, gluconate, glutamine, glycerophosphate, glycollylarsanilate, hemisulfate , heptanoate, hexanoate, hexylresorcinol salt, hydrabamate, bromide, chloride, 2-hydroxyethanesulfonate, hydroxynaphthoate, iodide, isonicotinate, isothiosulfate, lactate, lactobionate, laurate, apple acid salt, maleate, mandelate, methanesulfonate, methylnitrate, methylsulfate, methanesulfonate, mucate, 2-naphthalenesulfonate, naphthalenesulfonate, nicotinate, nitrate, oil acid salts, oxalates, pamoate (pamoate), palmitates, pantothenates, pectinates, persulfates, phosphates/hydrophosphates, pimelates, phenylpropionates, polygalacturonates, propionates, salicylates, stearates, sulfates, subacetates, succinates, tannates, tartrates, theocyanates, thiocyanates, toluenesulfonates , triethyl iodide, trifluoroacetates, undecanoates, valerates, and the like. In addition, if the compounds of the present invention have an acidic moiety, suitable pharmaceutically acceptable salts thereof include, but are not limited to, salts derived from inorganic bases, including aluminum salts, ammonium salts, calcium salts, copper salts, ferric salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts and the like. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, di- and tertiary amines, cyclic amines, dicyclohexylamines and alkaline ion exchange resins, for example, arginine, betaine, caffeine, choline, N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylamine, ethylenediamine, N-ethylmorpholine, N-ethylhexamethylenetetramine, reduced glucosamine, glucosamine, histidine, hydrazine, isopropylamine, lysine, methylreduced glucosamine, morpholine, hexahydropyrazine, hexahydropyridine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. Also included are basic nitrogen-containing groups that can be quaternized using reagents such as lower alkyl halides, such as the chlorides, bromides and iodides of methyl, ethyl, propyl and butyl; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and diamyl sulfate; long chain halides, such as the chlorides, bromides and iodides of decyl, lauryl, myristyl and stearyl; aralkyl halides, such as the bromides of benzyl and phenethyl, and others.

The composition provided by the present invention can be prepared by combining the active ingredient or composition provided by the present invention with at least one pharmaceutically acceptable carrier (vehicle) using the technology well known to those skilled in the art to which the present invention belongs, and preparing a dosage form suitable for the composition of the present invention. The dosage form includes but is not limited to solutions, emulsions, suspensions, powders, tablets, buccal tablets, pills, chewing gums, capsules and other dosage forms similar to or suitable for the present invention.

The "pharmaceutically acceptable carrier" in the present invention includes one or more types of ingredients selected from the following: solvents, emulsifiers, suspending agents, disintegrating agents, binders, shaping agents, stabilizers, chelating agents, diluents, gelling agents, preservatives, lubricants, surfactants, and other similar or applicable carriers in the present invention.

The above-mentioned composition may also be appropriately added with one or more dissolution aids, buffers, colorants, flavoring agents, etc. commonly used in the pharmaceutical preparation field as needed.

The "pharmaceutically acceptable excipient" in the present invention includes but is not limited to: at least one of a polymer, a resin, a plasticizer, a filler, a lubricant, a diluent, a binder, a disintegrant, a solvent, a co-solvent, a surfactant, a preservative, a sweetener, a flavoring agent, a pharmaceutical grade dye or pigment, and a viscosity agent.

The term "pharmaceutical composition" as used herein refers to a solid or liquid composition in a form, concentration and purity suitable for administration to a patient and which can induce the desired physiological changes after administration; the pharmaceutical composition is sterile and/or non-pyrogenic.

The "individual" in the present invention includes living organisms, such as humans, monkeys, cows, sheep, horses, pigs, cattle, goats, dogs, cats, mice, rats, cultured cells and transgenic species thereof. In a preferred embodiment, the individual is a human.

The term "administering" as used herein includes any route of administration that allows the active ingredient of the present invention to perform its intended function.

The terms "anti", "inhibit" and the like as used herein refer to preventing, delaying, improving, reducing or reversing the occurrence of symptoms.

"Colorectal cancer" as used herein refers to a general term for cancer originating from the colon or rectum, including but not limited to rectal cancer, colon-rectal cancer, colorectal cancer, intestinal cancer, hereditary non-polyposis colorectal cancer, etc. In the present invention, in the detection, analysis, classification or treatment, colorectal cancer-related cells include precancerous (e.g., benign), malignant, pre-metastatic, metastatic and non-metastatic cells.

The terms "treatment", "for treatment" and similar terms in the present invention are used herein to refer to the administration of a certain agent in order to obtain a certain effect. The effect is a therapeutic partial or complete cure of a disease and/or the symptoms of the disease. As used herein, "treatment" covers any treatment of colorectal cancer in mammals (especially humans), and includes: (a) inhibiting the disease, that is, preventing its development; and (b) alleviating the disease, that is, causing the disease to regress. In the treatment of tumors (such as colorectal cancer), therapeutic agents can directly reduce the growth and metastasis of tumor cells.

The term "prevention" as used herein means to inhibit or prevent symptoms associated with a target disease.

The "effective amount" in the present invention refers to the amount necessary to produce and cause the expected biological response, and is not the amount required for therapeutic cure. It is understood by those skilled in the art that the effective amount of the pharmaceutical composition may vary depending on factors such as the desired biological endpoint, the intended delivery of the biologically active agent, the composition of the encapsulating matrix, the target tissue, etc.

The terms "associated with" or "related to" and the like as used herein refer to the statistical correlation between two events, such events including numbers, data sets and the like. For example, when the events involve numbers, a positive correlation means that as one increases, the other also increases. A negative correlation means that as one increases, the other will decrease.

[Chalcone compounds containing selenium]

The selenium-containing chalcone compound of the present invention is represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

In formula (I), n is an integer from 0 to 4, and R is independently -OH, _-NH2 , -SH, -OR', _-NR'2 or -SR', wherein R' is a substituted or unsubstituted alkyl group; in the embodiment where R is _-NR'2 , multiple R's are the same or different and can be independently substituted or unsubstituted alkyl groups.

In the embodiment where n is 2, 3 or 4, multiple R's are the same or different from each other and can be independently -OH, _-NH2 , -SH, -OR', _-NR'2 or -SR'; wherein R' has the same meaning as above.

In one embodiment, n is preferably an integer from 0 to 4, and R is preferably -OH; in another embodiment, n is more preferably 0, and R is more preferably -OH. The selenium-containing chalcone compound of the above structure can further enhance the ability to inhibit colorectal cancer.

In one embodiment, the selenium-containing chalcone compound of the present invention is represented by any one of the following compounds: , , , , the ability to inhibit colorectal cancer can be further enhanced by using the selenium-containing chalcone compounds of the above structure.

The selenium-containing chalcone compounds of the present invention include all possible mirror image isomers, regioisomers and non-mirror image isomers and mixtures of two or more stereoisomers, such as mixtures of mirror image isomers and/or non-mirror image isomers in all ratios. Therefore, mirror image isomers are the subject of the present invention in the form of mirror image isomerically pure forms (both levorotatory mirror image and dextrorotatory mirror image), in the form of racemates, and in the form of mixtures of all ratios of the two mirror image isomers. In the case of cis/trans isomerism, the present invention includes cis forms and trans forms and mixtures of all ratios of these forms. If desired, the preparation of individual stereoisomers can be carried out by separation of mixtures by conventional methods (e.g. by chromatography or crystallization), by synthesis using stereochemically uniform starting materials or by stereoselective synthesis. Derivatization can be carried out before separation of stereoisomers, as appropriate. Separation of mixtures of stereoisomers can be carried out at an intermediate step during the synthesis of compounds of formula (I) or it can be carried out on the final racemic product. Absolute stereochemistry can be determined by X-ray crystallography of crystalline products or crystalline intermediates (derivatized, if necessary, with reagents containing stereocenters of known configuration). If the compounds of the invention are capable of isomeric interconversion, all individual isomeric interconverters and mixtures thereof are included within the scope of the invention. The present invention includes all such isomers, as well as salts, solvates (including hydrates) and solvates of such racemates, mirror isomers, non-mirror isomers and tautomeric isomers and mixtures thereof.

As used herein, whether or not particular abbreviations are specifically defined, the symbols and conventions used in the processes, schemes, and examples should be consistent with those used in the current scientific literature, for example, the Journal of the American Chemical Society or the Journal of Biological Chemistry. Specifically, but not limited to, the following abbreviations may be used in the Examples and throughout the specification: g (gram); mg (milligram); mL (milliliter); μL (microliter); mM (millimolar); M (micromolar); Hz (hertz); MHz (megahertz); mmol (millimolar); hr or hrs (hours); min (minutes); MS (mass spectrometry); ESI (electrospray ionization); and TLC (thin layer chromatography). For all of the following examples, standard work-up and purification methods known to those skilled in the art may be utilized. Unless otherwise indicated, all temperatures are expressed in °C (degrees Celsius). Unless otherwise indicated, all reactions were performed at room temperature. The synthetic methods explained herein are intended to illustrate applicable chemistry through the use of specific examples and are not indicative of the scope of the present disclosure.

[Pharmaceutical composition] The selenium-containing chalcone compound of the present invention can be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition, and its specific dosage form is not limited. For example, the dosage form prepared by the carrier can be a solution, suspension, emulsion, powder, tablet, pill, syrup, buccal tablet, tablet, chewing gum, slurry or capsule; the pharmaceutical composition can be packaged in a container and prepared into a test kit or product together with a package insert containing information related to the selenium-containing chalcone compound and the method of use.

To prepare the pharmaceutical composition of the present invention, one or more compounds of the present invention as active ingredients are mixed with a pharmaceutical carrier according to conventional pharmaceutical mixing techniques. The carrier can take a variety of forms depending on the desired preparation form for administration (e.g., oral or parenteral (e.g., intramuscular)). In preparing a composition in the form of an oral dosage form, any conventional pharmaceutical medium can be used. Thus, for liquid oral preparations (e.g., suspensions, elixirs and solutions), suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like; for solid oral preparations (e.g., powders, capsules, film-coated tablets, gelatin capsules and tablets), suitable carriers and additives include starch, sugar, diluents, granulating agents, lubricants, binders, disintegrants and the like. Tablets and capsules represent the most advantageous oral dosage unit form due to their ease of administration, in which case solid pharmaceutical carriers are obviously used. If desired, tablets may be coated with sugar or enteric coating by standard techniques. For parenteral administration, the carrier generally comprises sterile water, but other ingredients may be included, for example, for purposes such as aiding dissolution or for preservation. Injectable suspensions may also be prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed. The pharmaceutical compositions herein will contain an amount of active ingredient necessary to deliver an effective dose as described above per dosage unit (e.g., tablets, capsules, powders, injections, teaspoons and the like).

[Method for treating colorectal cancer, use of preparing a pharmaceutical composition for preventing or treating colorectal cancer] The present invention also discloses a method for treating colorectal cancer with a selenium-containing chalcone compound and use of preparing a pharmaceutical composition for preventing or treating colorectal cancer. The method is to administer a therapeutically effective amount of a selenium-containing chalcone compound as an active ingredient to a patient in need to prevent or treat colorectal cancer.

The treatment method of the present invention can be used in the treatment of colorectal cancer in mammalian subjects (especially humans). Subjects suffering from or at risk of developing tumors are all covered by the treatment method described herein.

The treatment method of the present invention is to administer a selenium-containing chalcone compound to an individual (e.g., a human patient) to inhibit the growth of colorectal cancer cells. The treatment method according to the present invention can also be used to reduce tumor size, reduce tumor burden and/or improve clinical outcomes in patients.

[Administration of selenium-containing chalcone compounds] Administration of selenium-containing chalcone compounds to colon cancer can be achieved by various methods, including intratumoral, intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, intraperitoneal, intraarterial, and rectal administration. Other suitable routes include administration of the composition orally, buccal, nasal, nasopharyngeal, parenteral, enteral, gastric, topical, transdermal, subcutaneous, intramuscular, in the form of tablets, solids, powders, liquids, aerosols, intralesional injection into the tumor, intralesional injection near the tumor, intravenous infusion, and intraarterial infusion. Local or systemic administration can be performed with or without the addition of a formulation. The drug can also be administered to the tumor site or around the tumor site of an individual through a slow release mode. The administration method is preferably selected from oral administration, injection, application, spray or patch to administer to the patient.

Liquid forms for oral or injection administration that may be incorporated into the novel compositions of the present invention include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils (e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical vehicles. Suitable dispersants or suspending agents for aqueous suspensions include synthetic and natural gums, such as tragacanth, gum arabic, alginates, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, or gelatin.

Tablets and capsules for oral administration are usually provided in unit dose form and contain customary formulators such as binders, fillers (including cellulose, mannitol, lactose), diluents, tableting agents, lubricants (including magnesium stearate), detergents, disintegrants (such as polyvinylpyrrolidone and starch derivatives such as sodium hydroxyacetate starch), colorants, flavor enhancers and wetting agents (such as sodium lauryl sulfate).

Solid compositions for oral administration can be prepared by conventional blending, filling or tableting methods. Blending operations can be repeated to distribute the active ingredient throughout the composition containing large amounts of fillers. Such operations are conventional.

For parenteral administration, a fluid unit dose containing the compound and a sterile vehicle may be prepared. Depending on the vehicle and concentration, the compound may be suspended or dissolved. Parenteral solutions are typically prepared by dissolving the compound in the vehicle, sterilizing by filtration, filling appropriate vials and sealing. Advantageously, adjuvants such as local anesthetics, preservatives and buffers may also be dissolved in the vehicle. To increase stability, the composition may be frozen after filling the vials and the water removed under vacuum. Parenteral suspensions are prepared in substantially the same manner, except that the compound may be suspended in the vehicle rather than dissolved and sterilized by exposure to ethylene oxide prior to suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent may be included in the composition to facilitate uniform distribution of the compound of the present application.

Pharmaceutical formulations for administration by inhalation may be delivered from a pressurized pack using an insufflator or nebulizer.

[Dosage] In the treatment method of the present invention, an effective amount of a selenium-containing chalcone compound is administered to an individual in need. Specifically, it varies depending on the purpose of administration, the health and physical condition of the individual to be treated, age, classification group of the individual to be treated (e.g., humans, non-human primates, primates, etc.), dosage form of the selenium-containing chalcone compound, the treating clinician's assessment of the medical situation, and other relevant factors. It is expected that the amount will be within a relatively wide range, which can be determined by routine tests. For example, in order to fully induce the death of colorectal cancer cells, the effective dose of the selenium-containing chalcone compound is preferably 10 μM to 200 μM, more preferably 10 μM to 100 μM, more preferably 10 μM to 46.95 μM, more preferably 10 μM to 38.23 μM, and most preferably 10 μM to 19.98 μM.

The dosage regimen utilizing the compound is selected based on a variety of factors, including the type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the patient's renal and hepatic function; and the specific compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progression of the condition.

[Preparation method of selenium-containing chalcone compounds] The preparation method of selenium-containing chalcone compounds of the present invention is to generate selenium-containing chalcone compounds by reacting acetophenone derivatives with selenophene-2-carbaldehyde via a Claisen-Schmidt reaction; wherein selenophene-2-carbaldehyde is obtained by adding DMF/POCl ₃ to selenophene, and the yield is 40~80%, preferably 51%.

In one embodiment, the preparation method of the selenium-containing chalcone compound of the present invention comprises a chemical reaction represented by the following formula (II):

In formula (II), n is an integer from 1 to 5, R ₁ is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ , -SR' or methoxy methyl (MOM); R ₂ corresponds to R ₁ and is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ or -SR', wherein R' is a substituted or unsubstituted alkyl group; in the embodiment where R ₁ and R ₂ are -NR' ₂ , multiple R's are the same or different and may be independently substituted or unsubstituted alkyl groups.

In the embodiment where n is 2, 3, 4 or 5, multiple R _1s are the same or different and can be independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ , -SR' or methoxymethyl ether, R ₂ corresponds to R ₁ and can be independently -OH, -NH 2 , -SH, -OR', -NR' ₂ or -SR'; wherein R' has the same meaning as above.

In one embodiment, n is an integer from 1 to 3, R ₁ is independently -OH or methoxymethyl ether, and R ₂ is -OH; in another embodiment, n is 1, R ₁ and R ₂ are -OH. The selenium-containing chalcone compound of the above structure can further enhance the ability to inhibit colorectal cancer.

In any embodiment where R ₁ is methoxymethyl ether, the preparation method of the present invention further comprises the step of subjecting the product of the reaction of formula (II) to an acid reaction with 3N HCl at 100 ^° C to convert the methoxymethyl ether into -OH; and in this embodiment, the condensation yield of the reaction is further improved.

In order to more clearly illustrate the preparation method of the present invention, the carbon atoms on the benzene ring of the acetophenone derivative are numbered as shown in the following figure:

In one embodiment, the structure of the acetophenone derivative A0 (hereinafter referred to as compound A0) is that n is 1, _R1 and _R2 are -OH and bonded to carbon 2, and the acetophenone derivative A0 and selenophene-2-carboxaldehyde are reacted in the formula (II); in this embodiment, no further acid reaction is required to obtain the selenium-containing chalcone compound A (hereinafter referred to as compound A) of the present invention. Compound A:

In one embodiment, the structure of the acetophenone derivative B0 (hereinafter referred to as compound B0) is n=2, wherein the first _R1 is -OH and is bonded to carbon No. 2, and the second _R1 is methoxymethyl ether and is bonded to carbon No. 4, and the acetophenone derivative B0 is reacted with selenophene-2-carboxaldehyde in the reaction of formula (II); in this embodiment, the product of the reaction of formula (II), compound B', is subjected to an acid reaction with 3N HCl at 100 ^° C to react the methoxymethyl ether on carbon No. 4 to -OH, that is, both _R2 are -OH and are respectively bonded to carbon No. 2 and carbon No. 4 to obtain the selenium-containing chalcone compound B of the present invention (hereinafter referred to as compound B). Compound B:

In one embodiment, the structure of the acetophenone derivative C0 (hereinafter referred to as compound C0) is n=2, wherein the first _R1 is -OH and is bonded to carbon No. 2, and the second _R1 is methoxymethyl ether and is bonded to carbon No. 5, and the acetophenone derivative C0 is reacted with selenophene-2-carboxaldehyde in the reaction of formula (II); in this embodiment, the product of the reaction of formula (II), compound C', is subjected to an acid reaction with 3N HCl at 100 ^° C to react the methoxymethyl ether on carbon No. 5 to -OH, that is, both _R2 are -OH and are respectively bonded to carbon No. 2 and carbon No. 5 to obtain the selenium-containing chalcone compound C of the present invention (hereinafter referred to as compound C). Compound C:

In one embodiment, the structure of the acetophenone derivative D0 (hereinafter referred to as compound D0) is that n is 3, wherein the first R ₁ is -OH and is bonded to carbon No. 2, the second R ₁ is methoxymethyl ether and is bonded to carbon No. 4, and the third R ₁ is methoxymethyl ether and is bonded to carbon No. 6, and the acetophenone derivative D0 is reacted with selenophene-2-carboxaldehyde in the reaction of formula (II); in this embodiment, the product of the reaction of formula (II), compound D', needs to be subjected to an acid reaction with 3N HCl at 100 ^° C, so that the methoxymethyl ether on carbon No. 4 and carbon No. 6 reacts to -OH, that is, the three R _2s are all -OH and are respectively bonded to carbon No. 2, No. 4, and No. 6 to obtain the selenium-containing chalcone compound D of the present invention (hereinafter referred to as compound D). Compound D:

The synthesis method of the present invention is very easy to operate, and the reactants can be easily obtained with moderate yield. The reactants are properly protected before synthesis. After synthesis, only the protecting group (MOM) needs to be removed by adding acid and then column chromatography or vacuum filtration is performed to obtain a pure selenium-containing chalcone compound.

[Preparation example and analysis method]

All key raw materials used in the present invention were purchased from various commercial sources (e.g., Aldrich and Alfa from Uni-Onward Corporation) and used without further purification. Purification was performed by flash column chromatography on 230-400 mesh SiO ₂ (SiliaFlash® P60, 40-63 μm 60 Å, SiliCycle® Inc.). The purity of each molecule was confirmed by 1H (600 MHz) and 13C (150 MHz) nuclear magnetic resonance (NMR) spectroscopy on a Bruker 600 MHz Ultrashield instrument. Molecular weights were measured by high resolution mass spectrometry (HRMS) by electrospray ionization (ESI) on a Bruker UltraFlex II instrument. Chemical shifts are reported in parts per million (ppm) with deuterated solvent residues as internal standards: 1H NMR (7.26 ppm for CDCl3 and 2.49 ppm for DMSO-d6); 13C NMR (77.0 ppm for _CDCl3 and 39.7 ppm for DMSO-d6). Melting points of synthesized compounds were determined by open capillary on MP-2D instrument and are uncorrected. The progress of the reaction was monitored by thin layer chromatography (TLC) on Analtech Silica gel HLF UV254 and observed by UV and stained with _KMnO4 or p-anisaldehyde solution.

General procedure for preparing selenophene-2-carboxaldehyde:

DMF (1.5618 g, 21.3691 mmol) was added to a round bottom flash, and POCl ₃ (1.6382 g, 10.6845 mmol) was slowly added under nitrogen at 4 ^° C. The mixture was stirred vigorously for 45 minutes, and then selenophene (0.7000 g, 5.3422 mmol) was added. The solution was gradually heated to 26 ^° C and stirred for 18 hours. At the end of the reaction, the mixture was quenched with NaHCO ₃ (saturated), sonicated for several minutes, and then extracted with Et ₂ O. The organic layer was separated, dried (MgSO4), concentrated and purified by azeotrope chromatography (hexane:EtOAc=7:1-6:1-3:1; hexane:EtOAc=4:1, _Rf =0.40) to give a yellow-green oil (0.4327 g, 2.7205 mmol) in 51% yield.

General procedure for preparing compound A, compound B', compound C', and compound D':

1 equivalent of compound A0, compound B0, compound C0, compound D0 and selenophene-2-carboxaldehyde (1.2 equivalents) were dissolved in ethanol (0.20 M), and then 40% NaOH aqueous solution (10 equivalents) was added at 26 ^° C and stirred for 17 hours. At the end of the reaction, 2N HCl was used to neutralize the reaction. The resulting mixture was added to distilled water and refrigerated for 8 hours. The precipitate was filtered, and washed with distilled water, hexane and dried in sequence.

General procedure for preparing compound B, compound C, and compound D:

Compound B', Compound C', and Compound D' were dissolved in THF:MeOH (1:1/v:v; 0.15 M), and then 3N HCl was added, and the mixture was heated to 100 ^° C. At the end of the reaction time, distilled water and _Et2O were added to the mixture. The organic layer was separated, dried ( _MgSO4 ), and purified by azeotropic column chromatography or by suction filtration to obtain the target molecules Compound B, Compound C, and Compound D, respectively.

The present invention is illustrated by the following embodiments. However, the following embodiments are merely illustrative of the present invention and should not be considered as limiting the scope of the present invention in any way.

[Cell culture]

Human colorectal adenocarcinoma cells (HT-29) were obtained from Yangming Jiaotong University. The cell line was cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μM L-glutamine, and 1 mM sodium pyruvate. The cells were cultured at 37°C and 5% CO ₂ and passaged twice a week. The cells were stored in liquid nitrogen at -155°C. After the cells were thawed, the experiments were completed before the 30th passage to reduce experimental errors. Culture-related compound stock solutions were stored at 10 mM in DMSO at -20°C and thawed immediately before use.

[Cell viability assay (methylthiazol tetrazolium assay, MTT)] Human colorectal adenocarcinoma cells (HT-29) were seeded in 96-well plates (5x10 ³ cells in 200 μl per well) and cultured for 24 hours. Compounds A to D were added at different concentrations (6.25, 12.5, 25, 50, 100 μM), and 5-fluorouracil (5-FU) was used as a positive control. After 72 hours of culture, 0.5 mg/ml of methylthiazol tetrazolium (MTT) (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma-Aldrich) was added and cultured at 37 ^o C for 3 hours. After removing the MTT reagent, DMSO was added and the cells were analyzed using an ELISA plate reader, TECAN Spark (Tecan Group Ltd.) (μ Quant) measured the OD value at a single wavelength of 570 nm and compared the cell survival to analyze the effects of each selenium-containing chalcone compound A~D on the growth of colorectal cancer cells. The results are shown in Table 1:

[Table 1] Selenium-containing chalcone compounds IC ₅₀ (μM) Compound A 19.98±3.38** Compound B 38.23±3.30* Compound C 46.95±5.68* Compound D >100 5-Fluorouracil (5-FU) 41.43±7.90* In Table 1, the values are presented as mean ± standard deviation (n=3); _IC50 indicates the drug concentration required to inhibit 50% of cells; * indicates p<0.05; ** indicates p<0.01; 5-fluorouracil is a known chemotherapy drug and serves as a positive control group.

As shown in Table 1, compound A exhibits the best inhibitory activity against human colorectal adenocarcinoma cells and is better than 5-FU, a conventional chemotherapy drug; compound B and compound C exhibit inhibitory activity comparable to that of 5-FU.

[Clonogenic assay]

Human colorectal adenocarcinoma cells (HT-29) were seeded at 3500 cells/well in 6-well cell culture plates and cultured for 24 hours. Then, different concentrations (5, 10, 20, 40 μM) of compound A were added, and 40 μM 5-fluorouracil (5-FU) was used as a positive control group, and DMSO was used as a negative control group. After 14 days of culture, the cells were washed 3 times with PBS, and 99% methanol was added to fix the cells for 30 minutes. After washing 3 times with distilled water, they were stained with 0.2% crystal violet solution and photographed. The cell coverage area was calculated using ImageJ (BioTechniques) for quantitative analysis to analyze the effects of different concentrations of compound A on the growth of colorectal cancer cells.

Please refer to FIG. 1 for the results. FIG. 1 shows the effect of a selenium-containing chalcone compound on the proliferation of human colorectal adenocarcinoma cells (HT-29) analyzed by a colony analysis method according to one embodiment of the present invention. The values are presented as mean ± standard deviation (n=3); * indicates p < 0.05 compared with the control group; *** indicates p < 0.001 compared with the control group. As can be seen from the figure, compound A reduces the formation of colonies in HT-29 cells in a dose-dependent manner. In other words, compound A can significantly reduce the formation of HT-29 colonies at 10 μM, and as the dose increases, the reduction in HT-29 colonies can be gradually increased at 20 μM and 40 μM, respectively.

[Western blot] Human colorectal adenocarcinoma cells (HT-29) (6x10 ⁵ cells) were inoculated into 6-cm culture dishes and allowed to reach 85%~90% confluence. Compound A at different concentrations (5, 10, 20, 40 μM) was then added. 40 μM 5-fluorouracil (5-FU) was used as a positive control group. Human colorectal adenocarcinoma cells (HT-29) were collected and subjected to radioimmunoprecipitation assay (RTA). The cells were lysed with RIPA buffer to obtain total protein lysate, separated by SDS-PAGE electrophoresis using 12.5% sodium dodecyl sulfate-polyacrylamide gel, and transferred to a PVDF membrane. After blocking, the membranes were incubated with specific primary antibodies (anti-bax, anti-Bcl-2 (Cell Signaling Inc., Danvers, MA, USA), anti-caspase-3, and anti-β-actin (GeneTex Inc., Irvine, CA, USA)) at 4°C overnight. The next day, the membranes were washed with 0.1% Tween 20 (TBST) in Tris-buffered saline and incubated at room temperature with horseradish peroxidase (HRP-contained). The membranes were incubated with specific secondary antibodies against HRP for 1 h at room temperature with shaking. After appropriate washing, each membrane was developed using an enhanced chemiluminescence (ECL) detection kit and the images were visualized using an ImageQuant LAS 4000 Mini biomolecular imager (GE Healthcare, MA, USA). The band density was quantified using ImageJ (BioTechniques, NY, USA).

Western blot analysis was used to analyze whether Compound A affects the expression of proteins related to apoptosis in human colorectal adenocarcinoma cells (HT-29), such as activation of Caspase 3 (activation from the precursor (Pro-casp3) state to the cleaved (Clv-casp3) state) as a biomarker of apoptosis, Bax as a protein that promotes apoptosis, and Bcl-2 as a protein that inhibits apoptosis; please refer to Figures 2 to 5, which respectively show the expression of pro-caspase 3 (Pro-casp3), cleaved-caspase 3 (Clv-casp3) and Bcl-2 (Clv-casp3) in human colorectal adenocarcinoma cells (HT-29) of each group in one embodiment of the present invention by immunoblotting analysis. 3 (Clv-casp3), Bax and Bcl-2 expression, * indicates p < 0.05 compared with the control group; ** indicates p < 0.01 compared with the control group. It can be seen from the figure that compound A inhibits the expression of Pro-casp3 (inactive state of Caspase 3), increases the expression of Clv-casp3 (active state of Caspase 3), promotes the expression of Bax, and inhibits the expression of Bcl-2; From the above results, it can be seen that compound A inhibits the growth of cancer cells by inducing apoptosis of HT-29 cells, and its pathway is dependent on the mechanism of mitochondria and caspase-3 (please refer to Figure 6).

[Spectral data]

Selenophene-2-carbaldehyde:

Yield: 51%. Purification by azeotropic column chromatography (hexane:EtOAc=7:1-6:1-3:1; hexane:EtOAc=4:1, R _f =0.40) gave a yellow-green liquid. ¹ H NMR (600 MHz, CDCl ₃ ) δ 9.80 (s, 1H), 8.48 (d, J = 5.4 Hz, 1H), 8.01 (t, J = 3.2 Hz, 1H), 7.46 (dd, J = 5.4, 3.9 Hz, 1H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 184.2, 150.2, 140.9, 139.4, 130.7.

Compound A: ( E )-1-(2-Hydroxyphenyl)-3-(selenophen-2-yl)prop-2-en-1- one

Yield: 42%. Mp 104.0-104.7 ^o C. Yellow powder (hexane:EtOAc=6:1, R _f =0.63). ¹ H NMR (600 MHz, CDCl ₃ ) δ 12.44 (s, 1H), 8.44 (d, J = 5.3 Hz, 1H), 8.14 (d, J = 8.2 Hz, 1H), 8.03 (d, J = 14.9 Hz, 1H), 7.87 (d, J = 3.4 Hz, 1H), 7.60 (d, J = 14.9 Hz, 1H), 7.54 (t, J = 7.1 Hz, 1H), 7.39 (dd, J = 5.3, 3.8 Hz, 1H), 6.98 (td, J = 7.0, 3.4 Hz, 1H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 192.8, 161.5, 145.6, 139.9, 137.4, 137.0, 136.1, 131.0, 130.7, 121.3, 121.0, 119.2, 117.7. HRMS (ESI) calculated for C ₁₃ H ₉ O ₂ Se [MH] ⁺ 276.9768. Found: 276.9776.

Compound B': ( E )-1-(2 - Hydroxy-4-(methoxymethoxy)phenyl)-3-(selenophen-2-yl)prop-2-en-1-one

Yield: 35%. Mp 79.2-79.9 ^o C. Orange powder (hexane:EtOAc=4:1, R _f =0.4). ¹ H NMR (600 MHz, CDCl ₃ ) δ 8.41 (d, J = 5.5 Hz, 1H), 8.15 (d, J = 9.0 Hz, 1H), 8.01 (d, J = 14.9 Hz, 1H), 7.83 (d, J = 3.6 Hz, 1H), 7.55 (d, J = 14.9 Hz, 1H), 7.38 (dd, J = 5.5, 3.8 Hz, 1H), 6.61 (dd, J = 9.0, 2.4 Hz, 1H), 6.55 (d, J = 2.4 Hz, 1H), 5.27 (s, 2H), 3.38 (s, 3H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 191.4, 164.8, 163.2, 145.6, 139.5, 137.2, 136.7, 132.6, 130.9, 120.6, 114.7, 108.3, 103.1, 93.7. HRMS (ESI) calculated for C ₁₅ H ₁₃ O ₄ Se [MH] ⁺ 336.9979. Found: 336.9989.

Compound B: ( E )-1-(2,4-Dihydroxyphenyl)-3-(selenophen-2-yl)prop-2-en-1- one

Yield: 65%. Mp 143.9-145.1 ^o C. Purification by azeotrope chromatography (hexane:EtOAc=9:1-6:1-1:1; hexane:EtOAc=4:1, R _f =0.40) gave an orange-yellow powder. ¹ H NMR (600 MHz, CDCl ₃ ) δ 13.37 (s, 1H), 10.71 (s, 1H), 8.41 (d, J = 5.5 Hz, 1H), 8.10 (d, J = 8.9 Hz, 1H), 7.99 (d, J = 14.9 Hz, 1H), 7.83 (d, J = 3.4 Hz, 1H), 7.54 (d, J = 14.9, 1H), 7.38 (dd, J = 5.4, 3.4 Hz, 1H), 6.41 (dd, J = 8.8, 2.2 Hz, 1H), 6.28 (d, J = 2.2 Hz, 1H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 3H 2 0 3 Se 3 N- 4 147.74 35.3 N- 4 147.74 35.3 N- 4 147.74 35.3 N- 4 147.74 35.3 N- 4 147.74 35.3 N- 4 147.74 _{35.3 N- 4} 147.74 ^35.3 N- 4 147.74 35.3 N- 4 147.74 35.3 N- 4 147.74

Compound C': ( E )-1-(2 - Hydroxy-5-(methoxymethoxy)phenyl)-3-(selenophen-2-yl)prop-2-en-1-one

Yield: 44%. Mp 62.2-63.5 ^o C. Filter and wash with water and hexane to obtain an orange solid (hexane:EtOAc=4:1, R _f =0.45). ¹ H NMR (600 MHz, CDCl ₃ ) δ 11.86 (s, 1H), 8.43 (d, J = 5.5 Hz, 1H), 7.99 (d, J = 15.0 Hz, 1H), 7.86 (d, J = 3.5 Hz, 1H), 7.67 (d, J = 2.9 Hz, 1H), 7.55 (d, J = 15.0 Hz, 1H), 7.39 (dd, J = 5.3, 5.3 Hz, 1H), 7.26 (dd, J = 8.9, 2.9 Hz, 1H), 6.94 (d, J = 8.9 Hz, 1H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 192.1, 156.1, 149.0, 145.6, 139.7, 137.3, 136.9, 130.9, 125.4, 121.9, 121.4, 118.5, 117.3, 94.9, 55.6. HRMS (ESI) calculated for C ₁₅ H ₁₃ O ₄ Se [MH] ⁺ 336.9979. Found: 336.9990.

Compound C: ( E )-1-(2,5 - Dihydroxyphenyl)-3-(selenophen-2-yl)prop-2-en-1-one

Yield: 86%. Mp177.1-178.2 ^o C. Brick red solid (hexane:EtOAc=4:1, R _f =0.25). ¹ H NMR (600 MHz, CDCl ₃ ) δ 11.67 (s, 1H), 9.17 (s, 1H), 8.41 (d, J = 5.5 Hz, 1H), 7.98 (d, J = 15.0 Hz, 1H), 7.84 (d, J = 3.5 Hz, 1H), 7.47 (d, J = 15.0 Hz, 1H), 7.39 (s, 1H), 7.38 (d, J = 5.6 Hz, 1H), 7.00 (dd, J = 8.8, 2.9 Hz, 1H), 6.83 (d, J = 8.8 Hz, 1H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 192.3, 154.3, 149.5, 145.7, 139.5, 136.9, 136.8, 131.0, 124.2, 121.7, 121.0, 118.4, 114.7. HRMS (ESI) calculated for C ₁₃ H ₉ O ₃ Se [MH] ⁺ 292.9717. Found: 292.9726.

Compound D': ( E )-1-(2 - Hydroxy-4,6-bis(methoxymethoxy)phenyl)-3-(selenophen-2-yl)prop-2-en-1-one

Yield: 50%. Mp 80.2-80.6 ^o C. Yellow powder (hexane:EtOAc=4:1, R _f =0.45). ¹ H NMR (600 MHz, CDCl ₃ ) δ 8.35 (d, J = 5.3 Hz, 1H), 7.80 (d, J = 15.2 Hz, 1H), 7.72 (d, J = 3.1 Hz, 1H), 7.36 (t, J = 4.9 Hz, 1H), 7.31 (d, J = 15.2 Hz, 1H), 6.26 (s, 1H), 6.24 (s, 1H), 5.28 (s, 2H), 5.22 (s, 2H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 191.9, _{397.0203 . ​} ^​

Compound D: ( E )-3-(Selenophen-2-yl ) -1-(2,4,6-trihydroxyphenyl)prop-2-en-1-one

Yield: 88%. Mp 134.8-136.1 ^o C. The solid was recrystallized in ether and hexane to give a brown powder (hexane:EtOAc=1:1, R _f =0.3). ¹ H NMR (600 MHz, CDCl ₃ ) δ 12.56 (s, 1H), 8.31 (d, J = 5.5 Hz, 1H), 7.89 (d, J = 15.1 Hz, 1H), 7.84 (d, J = 15.1 Hz, 1H), 7.69 (d, J = 3.7 Hz, 1H), 7.35 (dd, J = 5.5, 3.8 Hz, 1H), 5.84 (s, 2H). ¹³ C NMR (150 MHz, CDCl ₃ ) δ 190.9, 165.1, 164.5, 146.6, 137.2, 135.6, 135.2, 131.0, 127.3, 104.1, 95.0. HRMS (ESI) calculated for C ₁₃ H ₉ O ₄ Se [MH] ⁺ 308.9666. Found: 308.9676.

In summary, the present invention utilizes a simple synthesis method to condense hydroxyacetophenone derivatives substituted at different positions (Compound A0, Compound B0, Compound C0, Compound D0) with self-synthesized selenophene-2-carboxaldehyde to synthesize four selenium-containing chalcone compounds (Compound A, Compound B, Compound C, Compound D), and screens human colorectal adenocarcinoma cells (HT-29). It is found that compared with the known chemotherapy drug 5-FU, the four chalcone compounds have better therapeutic effects on colorectal cancer, among which Compound A has the most significant inhibitory effect on colorectal cancer.

The embodiments described above are only for illustrating the technical ideas and features of the present invention, and their purpose is to enable persons familiar with the art to understand the contents of the present invention and implement them accordingly. They cannot be used to limit the patent scope of the present invention. That is, any equivalent changes or modifications made based on the spirit disclosed by the present invention should still be included in the patent scope of the present invention.

without

FIG1 shows the effect of a selenium-containing chalcone compound on the proliferation of human colorectal adenocarcinoma cells (HT-29) analyzed by clonogenic assay according to one embodiment of the present invention; the upper portion of FIG1 shows the appearance of human colorectal adenocarcinoma cells growing in a culture dish, and the lower portion of FIG1 shows the quantitative results thereof.

FIG. 2 shows the expression level of pro-caspase 3 (Pro-casp3) in human colorectal adenocarcinoma cells (HT-29) of each group by immunoblot analysis according to one embodiment of the present invention.

FIG3 shows the expression level of cleaved-caspase 3 (Clv-casp3) in human colorectal adenocarcinoma cells (HT-29) of each group analyzed by immunoblotting according to one embodiment of the present invention.

FIG. 4 shows the expression level of Bax in human colorectal adenocarcinoma cells (HT-29) of each group analyzed by immunoblotting according to one embodiment of the present invention.

FIG. 5 shows the expression level of Bcl-2 in human colorectal adenocarcinoma cells (HT-29) of each group analyzed by immunoblotting according to one embodiment of the present invention.

FIG6 shows the possible mechanism by which the selenium-containing chalcone compound of one embodiment of the present invention induces apoptosis of human colorectal adenocarcinoma cells (HT-29).

without.

Claims (9)

A selenium-containing chalcone compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof: In formula (I), n is an integer from 0 to 4, and each R is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ or -SR', wherein each R' is independently a substituted or unsubstituted alkyl group. The selenium-containing chalcone compound as described in claim 1, wherein n is an integer from 0 to 2, and each R is independently -OH. The selenium-containing chalcone compound as described in claim 1, wherein the formula (I) is represented by any one of the following compounds: , , , . A pharmaceutical composition comprising the selenium-containing chalcone compound as described in any one of claims 1 to 3, and a pharmaceutically acceptable carrier. The pharmaceutical composition of claim 4, which is a solution, suspension, emulsion, powder, tablet, pill, syrup, buccal tablet, tablet, chewing gum, concentrate or capsule. A use of a selenium-containing chalcone compound for preparing a pharmaceutical composition for preventing or treating colorectal cancer, wherein the selenium-containing chalcone compound comprises the selenium-containing chalcone compound as described in any one of claims 1 to 3. The use as described in claim 6, wherein the effective dose of the selenium-containing chalcone compound is 10 μM to 200 μM. A method for preparing a selenium-containing chalcone compound comprises the steps of a chemical reaction represented by the following formula (II): In formula (II), n is an integer from 1 to 5, R ₁ is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ , -SR' or methoxymethyl ether; R ₂ is independently -OH, -NH ₂ , -SH, -OR', -NR' ₂ or -SR'; wherein R' is independently substituted or unsubstituted alkyl. The method for preparing a selenium-containing chalcone compound as described in claim 8, when any R ₁ is methoxymethyl ether, further comprises the step of reacting the product of the reaction of formula (II) with hydrochloric acid at 100 ^° C to convert the methoxymethyl ether into -OH.