Novel triazine derivative and pharmaceutical composition comprising same for preventing or treating cancer

Abstract

One aspect relates to a pharmaceutical composition comprising a novel triazine derivative for preventing or treating cancer, wherein the novel triazine derivative selectively inhibits GLUT3 in cancer cells, inhibits interaction between IGF1 and IGF1R, which are upper signaling mechanisms thereof, and significantly inhibits cancer cell glycolysis. The compound exhibits a remarkably excellent effect in the prevention, alleviation or treatment of cancer, and thus can be effectively used as a novel cancer therapeutic agent.

Description

Novel triazine derivatives and pharmaceutical compositions for the prevention or treatment of cancer containing the same

[0001] The invention relates to a novel triazine derivative and a pharmaceutical composition for the prevention or treatment of cancer containing the same.

[0002] According to a report by the World Health Organization (WHO), Koreans have the highest incidence rate of colorectal cancer among 184 countries worldwide, with 45 cases per 100,000 people. The number of colorectal cancer patients is expected to reach approximately 1.32 million, representing a 32% increase compared to 10 years ago. Currently, various attempts are being made to treat colorectal cancer by targeting specific characteristics of the disease, ranging from chemotherapy to targeted and immunotherapy drugs. However, there are limitations to sustained efficacy due to patient heterogeneity, metabolic adaptation issues in the tumor microenvironment, as well as existing side effects and resistance problems. Metabolic anticancer drugs are garnering attention as a new approach to overcome these challenges.

[0003] Metabolic anticancer drugs target metabolic specificities common to most cancers and are characterized by lower toxicity compared to conventional anticancer drugs. These advantages are being presented as an alternative to complement the limited response rates of targeted or immunotherapies. In fact, drugs such as Idhifa (enasidenib), Telaglenastat, OT-82, and AZD3965 have been reported, and in Korea, KAT-101, OKN-007, and Starvanib are currently under development. However, the only officially FDA-approved drug is Idhifa, a treatment for hematological cancers, and aside from asparaginase-based drugs used as adjuvant therapies, there are virtually no approved treatments available. Furthermore, while the efficacy of existing metabolic anticancer drugs has been proven primarily in hematological cancers, their effectiveness has not been sufficiently demonstrated in solid tumors such as colorectal cancer. Therefore, there is an urgent need to discover new biomarkers and develop treatments that target the heterogeneous metabolic characteristics of colorectal cancer.

[0004] In particular, research has been conducted on glucose transporters (GLUTs) related to glucose uptake among the metabolic characteristics of cancer cells, but conventional studies have mainly targeted GLUT1. GLUT1 is widely expressed in normal tissues as well, resulting in a lack of selectivity and consequently toxicity to normal cells.

[0005] In this invention, GLUT3 (glucose transporter 3) is proposed as a key biomarker to overcome these limitations. GLUT3 is a major factor mediating glucose metabolism and is known to be upregulated in various cancer types to promote tumor metabolism and proliferation.

[0006] However, although GLUT3-centered metabolic targeted therapy for solid tumors is a promising approach to address these unmet medical needs, therapeutic strategies that directly target GLUT3 have not been sufficiently developed. In particular, since no therapeutic strategies or drugs have been reported that selectively inhibit only GLUT3 among the various isoforms of GLUTs, there is a technological gap that can safely target tumor-specific metabolism.

[0007] Accordingly, the objective of the present invention is to provide a metabolic anticancer agent composition that regulates cancer metabolism by selectively inhibiting GLUT3, a specific metabolism that appears in cancer cells.

[0008] One aspect is to provide a compound represented by the following chemical formula 1 or a salt thereof:

[0009] [Chemical Formula 1]

[0010]

[0011] In the above chemical formula 1,

[0012] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0013] Another aspect is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[0014] [Chemical Formula 1]

[0015]

[0016] In the above chemical formula 1,

[0017] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0018] Another aspect is to provide a health functional food for the prevention or improvement of cancer comprising a compound represented by the following chemical formula 1 or a food-grade acceptable salt thereof:

[0019] [Chemical Formula 1]

[0020]

[0021] In the above chemical formula 1,

[0022] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0023] Another aspect provides a method for the prevention or treatment of cancer comprising the step of administering a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof to an individual in need thereof:

[0024] [Chemical Formula 1]

[0025]

[0026] In the above chemical formula 1,

[0027] R 1 to R 4Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0028] Another aspect is to provide the use of a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a drug for the prevention or treatment of cancer:

[0029] [Chemical Formula 1]

[0030]

[0031] In the above chemical formula 1,

[0032] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0033] One aspect provides a compound represented by the following chemical formula 1 or a salt thereof:

[0034] [Chemical Formula 1]

[0035]

[0036] In the above chemical formula 1,

[0037] R1 to R4 are each independently H, F, Cl, Br, I, or substituted or unsubstituted C1 to C6 alkyls.

[0038] The above compound or its salt may have one or more chiral carbon centers and accordingly may exist as R or S isomers, racemic mixtures, diastereomer mixtures, and individual diastereomers.

[0039] In addition, the compound represented by Chemical Formula 1 may include a hydrate or solvate of the compound. The hydrate and solvate may be prepared using known methods and are preferably non-toxic and water-soluble.

[0040] The term “salt” refers to a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base.

[0041] The term "Cm to Cn" (where m and n are each independently integers greater than or equal to 1, and n is greater than m in absolute value) means having m to n carbon atoms.

[0042] The term "alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group. For example, "C1 to C6 alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms. Specifically, "C1 to C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, tert-pentyl, etc. Additionally, the alkyl may be substituted or unsubstituted.

[0043] The term "substitution" refers to replacing a hydrogen atom within a molecular structure with a substituent so that the resulting compound is chemically stable without exceeding the valence of a specified atom. For example, "group A is substituted with substituent B" may mean that a hydrogen atom bonded to an atom, such as carbon, constituting the backbone of group A is replaced by substituent B, thereby forming a covalent bond between group A and substituent B.

[0044] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3 and R 4 Each can independently be a C1 to C4 alkyl.

[0045] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3and R 4 Each can independently be a C1 to C4 alkyl.

[0046] In one embodiment, the compound may be a compound represented by any one selected from the group consisting of the following chemical formulas 2 to 4:

[0047] [Chemical Formula 2]

[0048] ,

[0049] [Chemical Formula 3]

[0050] and

[0051] [Chemical Formula 4]

[0052] .

[0053] Specifically, the compound represented by Chemical Formula 1 or its salt can be prepared by the method shown in Reaction Scheme 1 or Reaction Scheme 2 below, but is not limited to preparation by such method. In particular, a person skilled in the art will fully understand that the compound represented by Chemical Formula 1 can be prepared by various methods using known techniques well known in the field.

[0054] The following reaction scheme 1 shows the step-by-step method for preparing a compound represented by chemical formula 2, which is a representative compound among the compounds represented by chemical formula 1.

[0055] [Reaction Equation 1]

[0056]

[0057] According to one aspect, a compound or a salt thereof produced by the above-described manufacturing method selectively inhibits GLUT3, inhibits the interaction between IGF1 and IGF1R, which are upstream signaling mechanisms, and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits a significantly superior effect in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized as a novel cancer treatment.

[0058] The following reaction scheme 2 shows the step-by-step method for preparing a compound represented by chemical formula 3, which is a representative compound among the compounds represented by chemical formula 1.

[0059] [Reaction Equation 2]

[0060]

[0061] According to one aspect, a compound or a salt thereof produced by the above-described manufacturing method selectively inhibits GLUT3 and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits a significantly superior effect in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized as a novel cancer treatment.

[0062] According to one aspect, a compound or a salt thereof produced by the above-described manufacturing method selectively inhibits GLUT3 and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits a significantly superior effect in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized as a novel cancer treatment.

[0063]

[0064] Another aspect provides a pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0065] [Chemical Formula 1]

[0066]

[0067] In the above chemical formula 1,

[0068] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0069] The above terms "alkyl," "salt," etc. may be within the aforementioned range.

[0070] The term "pharmaceuticalally acceptable" means exhibiting characteristics that are not toxic to cells or humans exposed to a specific compound according to one mode.

[0071] When the above compound contains relatively acidic functional groups, a base addition salt can be obtained by contacting a sufficient amount of base with the neutral form of the compound in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium, or similar salts. When the above compound contains relatively basic functional groups, an acid addition salt can be obtained by contacting a sufficient amount of acid with the neutral form of the compound in a pure solution or a suitable inert solvent. Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromide, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, or phosphoric acid, and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, souveric acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid, and further include salts of amino acids (e.g., arginine) and salts of organic acids such as glucuronic acid.

[0072] The term "prevention" may refer to any act of suppressing or delaying cancer in an individual through the administration of a pharmaceutical composition according to one aspect.

[0073] The term "treatment" may refer to any act in which the symptoms of cancer in an individual are improved or beneficially altered by the administration of a pharmaceutical composition according to one aspect.

[0074] Additionally, the above pharmaceutical composition may be provided as a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0075] Specifically, the carrier may be, for example, a colloidal suspension, powder, saline solution, lipid, liposome, microsphere, or nano-spherical particle. These may form a complex with or be associated with a transport means and may be transported in vivo using a transport system known in the art, such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation agents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption-enhancing substances, or fatty acids.

[0076] When the above pharmaceutical composition is formulated, it may be prepared using diluents or excipients such as commonly used lubricants, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration may include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms may be prepared by mixing at least one excipient with the above composition, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition, lubricants such as magnesium stearate and talc may also be used in addition to simple excipients. Liquid formulations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives, in addition to commonly used simple diluents like water and liquid paraffin. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions. Witepsol, macrogol, Tween 61, cacao oil, laurin oil, glycerogelatin, etc. may be used as bases for suppositories, and known diluents or excipients may be used when manufactured in the form of ophthalmic preparations.

[0077] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3 and R 4 Each can independently be a C1 to C4 alkyl.

[0078] In one embodiment, the compound may be a compound represented by any one selected from the group consisting of the following chemical formulas 2 to 4:

[0079] [Chemical Formula 2]

[0080] ,

[0081] [Chemical Formula 3]

[0082] and

[0083] [Chemical Formula 4]

[0084] .

[0085] In one embodiment, the cancer may have increased expression or activity of the GLUT (glucose transporter) gene or protein within the cancer cells.

[0086] The term "cancer" refers to a disease caused by cells that possess aggressive characteristics, such as dividing and growing beyond normal growth limits; invasive characteristics, such as infiltrating surrounding tissues; and metastatic characteristics, such as spreading to other parts of the body or outside.

[0087] The term "gene" may be used interchangeably with "polynucleotide" and refers to a nucleic acid (e.g., DNA or RNA, etc.) that codes for genetic information as a macromolecular substance formed by the combination of nucleotides. The polynucleotide may include all nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine, and uracil).

[0088] The term "protein" may be used interchangeably with "polypeptide" and "peptide" and refers to a polymer of amino acids of any length. The protein may include a modified amino acid polymer, for example, a polymer in a conjugated form with disulfide bond-forming, glycosylated, lipidized, or labeled components.

[0089] The aforementioned "GLUT (glucose transporter) protein" refers to a membrane-permeable protein that plays a role in transporting surrounding glucose into the cell.

[0090] In one embodiment, the GLUT may be one or more selected from the group consisting of GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13, and GLUT14, specifically one or more selected from the group consisting of GLUT1, GLUT2, GLUT3, GLUT4, and GLUT14, more specifically one or more selected from the group consisting of GLUT1, GLUT3, and GLUT4, and even more specifically GLUT3.

[0091] In one embodiment, the cancer is one selected from the group consisting of brain tumor, laryngeal cancer, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, gastric cancer, cholangiocarcinoma, colon cancer, breast cancer, triple negative breast cancer (TNBC), lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, pro-anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, kidney cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, spinal cord tumor, brainstem glioma, and pituitary adenoma. There may be more than one, specifically one or more selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and brain tumor, and more specifically, colorectal cancer.

[0092] In one embodiment, the compound or a pharmaceutically acceptable salt thereof may inhibit the expression or activity of the GLUT3 gene or protein.

[0093] The term "repression of expression" above includes the repression or downregulation of expression of a gene or protein, or modification to code for a protein that has lost its original function. This may refer to a direct modification of the DNA encoding the protein, a modification of the mRNA during post-transcriptional regulation, or a modification of the three-dimensional structure of the polypeptide during post-translational transcription. For example, a gene may be inactivated by removing part of the gene's coding sequence and / or regulatory sequence, or it may refer to a change in the function of the gene or the protein expressed by it resulting from structural modification of the polypeptide, dominant negative polypeptide expression, etc., caused by deletion of exon regions resulting from simultaneously targeting both intron regions surrounding one or more exon regions of the target gene.

[0094] The term "inhibition of activity" above means that a gene or protein is modified so that it cannot perform its original function. For example, it may refer to a change in the function of a gene or protein resulting from the expression or administration of a competitive inhibitor, phosphorylation of a polypeptide, etc.

[0095] In one embodiment, the compound or a pharmaceutically acceptable salt thereof may inhibit the interaction between IGF1 (insulin-like growth factor 1) and IGF1R (insulin-like growth factor 1 receptor).

[0096] In one embodiment, the inhibition of the interaction between IGF1 and IGF1R may be by one or more selected from the group consisting of inhibition of the expression or activity of the IGF1 gene or protein and inhibition of the expression or activity of the IGF1R gene or protein.

[0097] The inhibition of the interaction between the above-mentioned IGF1 and IGF1R includes, for example, the pharmaceutical composition binding to part or all of the IGF1 gene or protein to prevent IGF1 from binding to IGF1R, or the pharmaceutical composition binding to part or all of the IGF1R gene or protein to prevent IGF1 from binding to IGF1R, or modifying IGF1R to prevent signal transduction to downstream mechanisms.

[0098] In one embodiment, the inhibition of the activity of the IGF1R protein may be the inhibition of the phosphorylation of the IGF1R protein.

[0099] In one embodiment, the compound or a pharmaceutically acceptable salt thereof may inhibit glycolysis in cancer cells.

[0100] In one embodiment, the inhibition of the above-mentioned process may be one or more selected from the group consisting of inhibition of glucose uptake within cancer cells and inhibition of ATP production within cancer cells.

[0101] The above pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concomitantly used drugs, and other factors well known in the medical field. The administration may be given once a day or divided into several doses. For example, it may be given every other day or once a week. For example, it may be administered at 0.001 to 1000 mg / kg / day, specifically at 0.01 to 500 mg / kg / day, more specifically at 0.1 to 400 mg / kg / day, and even more specifically at 0.1 to 400 mg / kg / day, 0.1 to 300 mg / kg / day, 0.1 to 250 mg / kg / day, 0.1 to 100 mg / kg / day, 0.1 to 10 mg / kg / day, 0.1 to 1 mg / kg / day, 1 to 400 mg / kg / day, 1 to 300 mg / kg / day, 1 to 250 mg / kg / day, 1 to 100 mg / kg / day, 1 to 10 mg / kg / day, 10 to It may be administered at 400 mg / kg / day, 10 to 300 mg / kg / day, 10 to 250 mg / kg / day, 10 to 100 mg / kg / day, 100 to 400 mg / kg / day, 100 to 300 mg / kg / day, 100 to 250 mg / kg / day, 250 to 400 mg / kg / day, 250 to 300 mg / kg / day, or 300 to 400 mg / kg / day.

[0102] The term "administration" above means introducing a specific substance into an individual by an appropriate method, and "individual" refers to all living organisms, including rats, mice, pigs, horses, cattle, and livestock, including humans capable of harboring cancer. Specific examples may include mammals, including humans.

[0103] The above pharmaceutical composition may be administered orally or parenterally, and when administered parenterally, a method of injection may be selected, such as external application to the skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intra-arterial injection, intramedullary injection, intracardiac injection, intrathecal injection, transdermal injection, nasal injection, enteral injection, local injection, sublingual injection, rectal injection, or thoracic injection.

[0104] In one embodiment, the pharmaceutical composition may further include other cancer therapeutic agents in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof, and the other cancer therapeutic agent may be an IGF1R inhibitor. For example, the other cancer treatments mentioned above may be linsitinib, NVP-AEW541, GSK1904529A, NVP-ADW742, BMS-536924, ceritinib, picropodophyllin, brigatinib, teprotumumab, ganitumab, figitumumab, cixutumumab, dalotuzumab, R1507, XL-228, INSM-18, BMS-754807, AG-1024, GSK1838705A, PQ 401, etc., and specifically may be linsitinib.

[0105] The above "IGF1R inhibitor" refers to a polynucleotide, polypeptide, small molecule compound, etc. that can inhibit signal transduction via IGF1R by targeting the IGF1R gene or protein, which is known to be involved in cell differentiation, growth, and apoptosis as a transmembrane tyrosine kinase receptor and is overexpressed in various cancer cells.

[0106] The above "lincitinib" is a compound represented by the following chemical formula 5, and is a potent, selective, and orally bioavailable dual inhibitor of the IGF1 receptor and the insulin receptor (IR). Lincitinib exerts an antitumor effect by inhibiting the ligand-dependent autophosphorylation of the tyrosine kinase receptors and inhibiting downstream signal transduction:

[0107] [Chemical Formula 5]

[0108] .

[0109] The other cancer therapeutic agent mentioned above may be included in the pharmaceutical composition in the minimum amount to obtain maximum effect without side effects, which can be easily determined by a person skilled in the art.

[0110] In one embodiment, the pharmaceutical composition may be administered in combination with other cancer treatments. That is, the pharmaceutical composition may be administered in combination with other cancer treatments, simultaneously, separately, or sequentially, and may be administered as a single or multiple doses. For example, the pharmaceutical composition may be administered in combination with an IGF1R inhibitor. Specifically, it may be administered in combination with lincitinib.

[0111] The term "concurrent administration" above means administering two or more drugs simultaneously or during the same treatment period.

[0112] When the above-mentioned other cancer therapeutic agents or IGF1R inhibitors are administered in combination with the above-mentioned pharmaceutical composition, they may be administered in an amount or ratio that can obtain maximum effect without side effects, and this can be easily determined by a person skilled in the art. For example, a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof and an IGF1R inhibitor may be administered at a concentration ratio of 10000:1 to 1:10000, specifically 10000:1, 1000:1, 100:1, 50:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:50, 1:100, 1:100, 1:100, 1:1000, or 1:10000, specifically 10000:1, 1000:1, 100:1, 50:1, 10:1, 5:1, or 1:1.

[0113] When the above pharmaceutical composition further includes the above other cancer treatment or IGF1R inhibitor, or when the above pharmaceutical composition is administered in combination with the above other cancer treatment or IGF1R inhibitor, a synergistic effect may occur in which the cell viability of cancer cells is significantly reduced compared to when only the above pharmaceutical composition is administered, resulting in a significantly superior effect for the prevention or treatment of cancer. Furthermore, in the case of the above synergistic effect, if the dosage of the compound represented by Formula 1 or its pharmaceutically acceptable salt is higher than that of lincitinib, the cell viability of cancer cells is reduced more significantly.

[0114] The other cancer treatments or IGF1R inhibitors mentioned above may be conventionally known cancer treatments or IGF1R inhibitors or newly developed cancer treatments or IGF1R inhibitors.

[0115] In one embodiment, it was confirmed that the expression level of GLUT3 increases in colorectal cancer patients, and that GLUT3 affects the regulation of glycolysis in cancer cells and influences cell proliferation and growth. Additionally, it was confirmed that IGF1R regulates the activity of GLUT3 as a upstream mechanism. Accordingly, the utility of a compound targeting the interaction between IGF1R and GLUT3 as an anticancer agent was confirmed (see Experimental Example 1).

[0116] In another example, it was confirmed that the 93rd hit compound (SGT-24093) exhibits excellent efficacy as an inhibitor of GLUT3. Furthermore, it was confirmed that SGT-24093 significantly reduces the cell viability of cancer cells, inhibits glucose uptake and ATP production within cancer cells, and selectively inhibits only GLUT3. Additionally, it was confirmed that SGT-24093 regulates the activity of GLUT3 by inhibiting IGF1R signaling. Accordingly, it was confirmed that SGT-24093 can be usefully employed as a pharmaceutical composition for the prevention or treatment of cancer (see Experimental Examples 2 to 4).

[0117] In another example, it was confirmed that when SGT-24093 was co-treated with lincitinib, known as an IGF1R inhibitor, the cell viability of cancer cells decreased as the treatment concentrations of SGT-24093 and lincitinib increased, and a synergistic effect occurred as the cell viability of cancer cells decreased significantly compared to when only a single compound was treated (see Experimental Example 5).

[0118] In another example, it was confirmed that SGT-24-103, a compound represented by Chemical Formula 3, exhibits excellent efficacy as an inhibitor of GLUT3. Furthermore, it was confirmed that compound SGT-24-103 does not exhibit cytotoxicity to normal cells while significantly reducing the cell viability of cancer cells, inhibiting glucose uptake and ATP production within cancer cells, and selectively inhibiting only GLUT3. Additionally, compound SGT-24-103 showed inhibitory effects on cancer cell growth not only in colorectal cancer but also in gastric cancer, liver cancer, lung cancer, breast cancer, prostate cancer, and glioblastoma, confirming its inhibitory activity against various types of cancer. Accordingly, it was confirmed that compound SGT-24-103 can be usefully employed as a pharmaceutical composition for the prevention or treatment of cancer (see Experimental Example 6).

[0119] A pharmaceutical composition according to one aspect selectively inhibits GLUT3, inhibits the interaction of IGF1 and IGF1R, which are upstream signaling mechanisms, and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits a significantly superior effect in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized as a novel cancer treatment.

[0120]

[0121] Another aspect provides a health functional food for the prevention or improvement of cancer comprising a compound represented by the following chemical formula 1 or a food-grade acceptable salt thereof:

[0122] [Chemical Formula 1]

[0123]

[0124] In the above chemical formula 1,

[0125] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0126] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3 and R 4 Each can independently be a C1 to C4 alkyl.

[0127] In one embodiment, the compound may be a compound represented by any one selected from the group consisting of the following chemical formulas 2 to 4:

[0128] [Chemical Formula 2]

[0129] ,

[0130] [Chemical Formula 3]

[0131] and

[0132] [Chemical Formula 4]

[0133] .

[0134] The above terms "alkyl," "salt," "prevention," etc. may be within the aforementioned range.

[0135] The term "improvement" may refer to any action that at least reduces parameters related to the condition being treated, such as the severity of symptoms. In this case, the health functional food may be used for the prevention or improvement of cancer, either simultaneously with or separately from a therapeutic drug, either before or after the onset of the disease.

[0136] The above "health functional(s) food" includes health functional foods, health foods, and health supplements.

[0137] The term "functional food" is synonymous with "food for special health use (FosHU)," and refers to a food with high medical or health effects that is processed to efficiently exhibit bio-regulatory functions in addition to providing nutrition. Here, "functional" means obtaining useful effects for health purposes, such as regulating nutrients or physiological actions regarding the structure and function of the human body.

[0138] In addition, "health food" refers to food that has active effects on maintaining or promoting health compared to general food, and "health supplement food" refers to food intended for the purpose of supplementing health.

[0139] In the above-mentioned health functional food, the compound represented by Chemical Formula 1 or its food-grade acceptable salt may be added directly to the food or mixed with other foods or food ingredients and used appropriately according to conventional methods. The amount of the compound represented by Chemical Formula 1 or its food-grade acceptable salt mixed may be appropriately determined according to its intended use (for the prevention or improvement of cancer). Generally, when manufacturing food or beverages, the compound represented by Chemical Formula 1 or its food-grade acceptable salt may be added in an amount of about 15% by weight or less, specifically about 10% by weight or less, with respect to the raw materials. However, in the case of long-term consumption for the purpose of health and hygiene or for health control, the above amount may be less than the above range.

[0140] The above-mentioned health functional food may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, by further including one or more of a carrier, a diluent, an excipient, and an additive. Foods to which a compound according to one aspect may be added include various food products, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, etc.

[0141] Specific examples of the above carrier, excipient, diluent, and additive may be one or more selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0142] The above-mentioned health functional food may contain other ingredients as essential components without special limitations, in addition to containing the compound represented by Chemical Formula 1 or a food-grade acceptable salt thereof. For example, it may contain various flavoring agents or natural carbohydrates as additional ingredients, such as in ordinary beverages. Examples of the above-mentioned natural carbohydrates may be monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, etc.; polysaccharides, e.g., dextrin, cyclodextrin, etc., common sugars; and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used. The proportion of the above-mentioned natural carbohydrates may be appropriately determined by the choice of a person skilled in the art.

[0143] In addition to the above, a health functional food according to one aspect may contain various nutritional supplements, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These ingredients may be used independently or in combination, and the proportion of these additives may also be appropriately selected by a person skilled in the art.

[0144] In one embodiment, the cancer may have increased expression or activity of the GLUT (glucose transporter) gene or protein within the cancer cells.

[0145] In one embodiment, the cancer may be one or more selected from the group consisting of brain tumor, laryngeal cancer, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, stomach cancer, colon cancer, breast cancer, triple negative breast cancer (TNBC), lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, pro-anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, kidney cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, spinal cord tumor, brainstem glioma and pituitary adenoma. Specifically, it may be one or more selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and brain tumor, and more specifically, it may be colorectal cancer.

[0146] In one embodiment, the compound or a food-grade acceptable salt thereof may inhibit the expression or activity of the GLUT3 gene or protein.

[0147] In one embodiment, the compound or a food-grade acceptable salt thereof may inhibit the interaction between IGF1 (insulin-like growth factor 1) and IGF1R (insulin-like growth factor 1 receptor).

[0148] In one embodiment, the inhibition of the interaction between IGF1 and IGF1R may be by one or more selected from the group consisting of inhibition of the expression or activity of the IGF1 gene or protein and inhibition of the expression or activity of the IGF1R gene or protein.

[0149] In one embodiment, the compound or a food-grade acceptable salt thereof may inhibit glycolysis in cancer cells.

[0150] In one embodiment, the health functional food may further include other health functional foods for the prevention or improvement of cancer in addition to the compound or a food-grade acceptable salt thereof. The other health functional food for the prevention or improvement of cancer may be an IGF1R inhibitor.

[0151] The above-mentioned health functional food for the prevention or improvement of other cancers may be included in the above-mentioned health functional food in a minimum amount that can obtain maximum effect without side effects, and this can be easily determined by a person skilled in the art.

[0152] In addition, in one embodiment, the health functional food may be consumed alone or in combination with the health functional food for the prevention or improvement of cancer. That is, the health functional food may be consumed in conjunction with other health functional foods for the prevention or improvement of cancer, and may be consumed simultaneously, separately, or sequentially, and may be consumed as a single or multiple foods. For example, the health functional food may be consumed in combination with an IGF1R inhibitor.

[0153] When the above health functional food further includes the above other health functional food, or when the above health functional food is consumed in combination with the above other health functional food, a synergistic effect may occur in which the effect of preventing or improving cancer becomes significantly superior, such as by significantly reducing the cell survival rate of cancer cells compared to when the above health functional food is consumed alone.

[0154] When consumed in combination with the above-mentioned health functional food for the prevention or improvement of cancer or an IGF1R inhibitor, it may be consumed in an amount or ratio that can obtain maximum effect without side effects, and this can be easily determined by a person skilled in the art. For example, a compound represented by Chemical Formula 1 or a food-grade salt thereof and an IGF1R inhibitor may be consumed in a concentration ratio of 10,000:1 to 1:10,000, specifically 10,000:1, 1,00:1, 50:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:50, 1:100, 1:100, 1:100, or 1:10,000, specifically 10,000:1, 1,000:1, 50:1, 10:1, 5:1, or 1:1.

[0155] The above-mentioned other health functional foods for the prevention or improvement of cancer may be conventionally known health functional foods for the prevention or improvement of cancer or newly developed health functional foods for the prevention or improvement of cancer.

[0156] A health functional food according to one aspect selectively inhibits GLUT3, inhibits the interaction of IGF1 and IGF1R, which are upstream signaling mechanisms, and significantly inhibits glycolysis in cancer cells. That is, since the health functional food inhibits the proliferation, growth, and development of cancer cells and exhibits significantly superior effects in the prevention, improvement, or treatment of cancer, it can be effectively utilized as a new health functional food for the prevention or improvement of cancer.

[0157]

[0158] Another aspect provides a method for the prevention or treatment of cancer comprising the step of administering a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof to an individual in need thereof:

[0159] [Chemical Formula 1]

[0160]

[0161] In the above chemical formula 1,

[0162] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0163] The above terms "alkyl," "salt," "prevention," "treatment," etc. may be within the aforementioned scope.

[0164] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3 and R 4 Each can independently be a C1 to C4 alkyl.

[0165] In one embodiment, the compound may be a compound represented by any one selected from the group consisting of the following chemical formulas 2 to 4:

[0166] [Chemical Formula 2]

[0167] ,

[0168] [Chemical Formula 3]

[0169] and

[0170] [Chemical Formula 4]

[0171] .

[0172] In one embodiment, the cancer may be one or more selected from the group consisting of brain tumor, laryngeal cancer, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, stomach cancer, colon cancer, breast cancer, triple negative breast cancer (TNBC), lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, pro-anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, kidney cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, spinal cord tumor, brainstem glioma and pituitary adenoma. Specifically, it may be one or more selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and brain tumor, and more specifically, it may be colorectal cancer.

[0173] In one embodiment, the pharmaceutical composition may further include other cancer therapeutic agents in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof, and the other cancer therapeutic agent may be an IGF1R inhibitor.

[0174] In one embodiment, the method for preventing or treating cancer may further include the step of administering another cancer treatment agent. That is, the pharmaceutical composition may be administered in conjunction with another cancer treatment agent, simultaneously, separately, or sequentially, and may be administered as a single or multiple doses. For example, the pharmaceutical composition may be administered in combination with an IGF1R inhibitor.

[0175] A compound according to one aspect selectively inhibits GLUT3, inhibits the interaction of its upstream signaling mechanisms, IGF1 and IGF1R, and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits significantly superior effects in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized in cancer treatment methods.

[0176]

[0177] Another aspect provides the use of a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a drug for the prevention or treatment of cancer:

[0178] [Chemical Formula 1]

[0179]

[0180] In the above chemical formula 1,

[0181] R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

[0182] The above terms "alkyl," "salt," "prevention," "treatment," etc. may be within the aforementioned scope.

[0183] In one embodiment, the above R 1 and R 2 is each independently H, F, Cl, Br, or I; and the above R 3 and R 4Each can independently be a C1 to C4 alkyl.

[0184] In one embodiment, the compound may be a compound represented by any one selected from the group consisting of the following chemical formulas 2 to 4:

[0185] [Chemical Formula 2]

[0186] ,

[0187] [Chemical Formula 3]

[0188] and

[0189] [Chemical Formula 4]

[0190] .

[0191] In one embodiment, the cancer may be one or more selected from the group consisting of brain tumor, laryngeal cancer, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, stomach cancer, colon cancer, breast cancer, triple negative breast cancer (TNBC), lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, pro-anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, kidney cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, spinal cord tumor, brainstem glioma and pituitary adenoma. Specifically, it may be one or more selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and brain tumor, and more specifically, it may be colorectal cancer.

[0192] In one embodiment, the pharmaceutical composition may further include other cancer therapeutic agents in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof, and the other cancer therapeutic agent may be an IGF1R inhibitor.

[0193] In one embodiment, the compound for manufacturing the drug may be administered in combination with another anticancer agent. That is, the compound may be administered in parallel with another anticancer agent, simultaneously, separately, or sequentially, and may be administered as a single or multiple doses. For example, the compound may be administered in combination with an IGF1R inhibitor.

[0194] A compound according to one aspect selectively inhibits GLUT3, inhibits the interaction of its upstream signaling mechanisms, IGF1 and IGF1R, and significantly inhibits glycolysis in cancer cells. That is, by significantly inhibiting the proliferation, growth, and development of cancer cells, it exhibits significantly superior effects in the prevention, improvement, or treatment of cancer, and thus can be effectively utilized as a novel cancer treatment.

[0195] According to one aspect, a novel triazine derivative selectively inhibits GLUT3 in cancer cells, inhibits the interaction of IGF1 and IGF1R, which are upstream signaling mechanisms, and significantly inhibits glycolysis in cancer cells. That is, the compound significantly inhibits the proliferation, growth, and development of cancer cells, thereby exhibiting a significantly superior effect in the prevention, improvement, or treatment of cancer, and thus can be usefully utilized as a novel cancer treatment.

[0196] Figure 1 shows the results of analyzing a bioinformatics database of colorectal cancer patient tissues.

[0197] Figure 2 shows GLUT3 mRNA expression in colorectal cancer patient tissue.

[0198] Figure 3 shows the results of immunohistochemistry (IHC) analysis in colorectal cancer patient tissue.

[0199] Figure 4 shows the expression of GLUT3 protein in colorectal cancer patient tissue.

[0200] Figure 5 shows the results of a Kaplan-Meier survival rate analysis using the TCGA dataset.

[0201] Figure 6 shows the expression of GLUT1 to 14 mRNA in colorectal cancer patient tissue.

[0202] Figure 7 is a confocal microscope image confirming GLUT3 expression after treating SW620 cells with shControl or shGLUT3.

[0203] Figure 8 shows the number of cells when treated with shControl or shGLUT3 (left) and when GLUT3 was overexpressed (right).

[0204] Figure 9 shows the colony formation of cells when treated with shControl or shGLUT3 by performing colony formation and anchorage-independent growth assays.

[0205] Figure 10 shows the colony formation and anchorage-independent growth assays performed to confirm the anchorage-independent growth of cells when treated with shControl or shGLUT3.

[0206] Figure 11 is a figure showing the effect of GLUT3 on the cell cycle by performing flow cytometry analysis with shControl or shGLUT3 treatment.

[0207] Figure 12 is a figure confirming glucose uptake in cells treated with shControl or shGLUT3.

[0208] Figure 13 is a figure confirming ATP production in cells treated with shControl or shGLUT3.

[0209] Figure 14 is a figure confirming glucose uptake in cells with a control vector or GLUT3 overexpression.

[0210] Figure 15 is a figure confirming ATP production in cells with a control vector or GLUT3 overexpression.

[0211] Figure 16 is a figure evaluating tumor growth in a subcutaneous xenograft model treated with shControl or shGLUT3.

[0212] Figure 17 shows the results of Gene Set Enrichment Analysis (GSEA) on RNA-seq data of colorectal cancer patient samples with increased GLUT3 expression.

[0213] Figure 18 is a figure confirming the upstream mechanism regulating GLUT3 expression.

[0214] Figure 19 is a figure confirming the correlation between IGF1 mRNA and GLUT3 mRNA expression.

[0215] Figure 20 shows the results of performing immunohistochemistry (IHC) on tissue samples from colorectal cancer patients with high IGF1 expression levels.

[0216] Figure 21 shows the results of RNA sequencing (RNA-Seq) analysis in tissue samples from colorectal cancer patients with high expression levels.

[0217] Figure 22 shows the results of a Kaplan-Meier survival rate analysis using the TCGA dataset of colorectal cancer patients.

[0218] Figure 23 shows the levels of GLUT3 protein in cells when treated with si-Control or si-IGF1R.

[0219] Figure 24 shows glucose uptake and ATP production in cells when treated with si-Control or si-IGF1R.

[0220] Figure 25 shows the results of screening GLUT3 selective inhibitors.

[0221] Figure 26 shows the GLUT3 protein expression when each of the eight compounds was treated.

[0222] Figure 27 shows the glucose uptake in cells when each of the eight compounds was treated.

[0223] Figure 28 shows the ATP production in cells when each of the eight compounds was treated.

[0224] Figure 29 shows the cell viability when each of the eight compounds was treated.

[0225] Figure 30 is a schematic diagram showing the synthesis process of compound SGT-24-104.

[0226] Figure 31 is a figure evaluating the cell viability of IEC6 and HCT116 cells, respectively, when treated with SGT-24093 and SGT-24-101.

[0227] Figure 32 shows the glucose uptake of cells when SGT-24093 and SGT-24-101 were treated to HCT116.

[0228] Figure 33 shows the ATP production of cells when SGT-24093 and SGT-24-101 were treated to HCT116.

[0229] Figure 34 shows the cell viability of IEC6 and HCT116 when SGT-24093 was treated.

[0230] Figure 35 shows the IC50 of SGT-24093 measured in HCT116 cells.

[0231] Figure 36 shows the GLUT3 luciferase activity measured when treated with SGT-24093.

[0232] Figure 37 is a figure confirming the expression of GLUT1, GLUT3, and GLUT4 when treated with SGT-24093.

[0233] Figure 38 shows the glucose uptake of cells when treated with SGT-24093.

[0234] Figure 39 shows the ATP production of cells when treated with SGT-24093.

[0235] Figure 40 shows the expression of p-IGFR1, IGFR1, and GLUT3 when treated with lincitinib.

[0236] Figure 41 shows the cell viability evaluated when SGT-24093 and lincitinib were administered in combination.

[0237] Figure 42 shows the screening results of GLUT3 selective inhibitors.

[0238] Figure 43 shows the evaluation of the stability in normal colon cells of 10 compounds (27, 28, 40, 92, 93, 101, 102, SGT-24-103, 104, 105) selected for showing the top 30% cancer inhibition rate in screening results.

[0239] Figure 44 is a figure evaluating the cancer inhibition rate by treating colon cancer cells with the 10 selected compounds mentioned above.

[0240] Figure 45 is a figure evaluating the effect of the 10 selected compounds on glucose metabolism in colon cancer cells.

[0241] Figure 46 is a figure evaluating the effect of the 10 selected compounds on ATP production capacity in colon cancer cells.

[0242] Figure 47 shows the IC50 values ​​of the GLUT3 inhibitor SGT-24-103 for various colorectal cancer cell lines.

[0243] Figure 48 shows the glucose metabolic capacity and ATP production inhibition rate according to the concentration of SGT-24-103 in colon cancer cells.

[0244] Figure 49 shows the degree of inhibition of tumor colony formation in colorectal cancer cells according to the concentration of SGT-24-103 treatment.

[0245] Figure 50 is a figure showing that SGT-24-103 does not affect the expression of other isoform GLUT proteins such as GLUT1 and GLUT4.

[0246] Figures 51 to 54 show the cancer-suppressing ability of SGT-24-103 in cell lines of gastric cancer (Fig. 51), liver cancer (Fig. 52), lung cancer (Fig. 53), breast cancer (MDA-MB-231 in Fig. 54), prostate cancer (LuCAP in Fig. 54), and glioblastoma (A172 in Fig. 54).

[0247] Figure 55 is a schematic diagram showing the synthesis process of compound SGT-24-103.

[0248] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0249]

[0250] Reference Example

[0251] <Reference Example 1> Bioinformatics Database Analysis

[0252] The GSE39582 dataset was obtained from the GEO database (https: / / www.ncbi.nlm.nih.gov / geo / ), and the TCGA database and IHC expression analysis of GLUT3 were obtained from the Human Protein Atlas (https: / / www.proteinatlas.org / ). The MGC9454 dataset was obtained from Kmplot (https: / / www.kmplot.com / ), and biological process analysis was obtained from Shiny GO (http: / / bioinformatics.sdstate.edu / go / ). In addition, data obtained from publicly available online analysis platforms such as muTARGET (https: / / www.mutarget.com / ), TNM Plot (https: / / tnmplot.com / analysis / ), and GEPIA (http: / / gepia.cancer-pku.cn / ) were utilized.

[0253]

[0254] <Reference Example 2> Cell Culture and Cell Proliferation

[0255] HCT116 (ATCC, #CCL-247) and SW620 (ATCC, #CCL-227) cells were purchased from the American Type Culture Collection, and all cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (HyClone) supplemented with 10% (v / v) FBS (Thermo Fisher Scientific), 100 U / ml penicillin, and 100 μg / ml streptomycin (Thermo Fisher Scientific). HumanKine ® Recombinant human IGF-1 protein was purchased from Humanzyme and used after diluting in PBS.

[0256] For the cell proliferation assay, 1 × 10⁶ SW620 cells transformed with shControl or shGLUT3 were placed in a 6-well plate. 5Cells were seeded at a density of cells / well. For HCT116 cells transformed with the control vector or GLUT3 / pEJ-3HA, 1 × 10⁶ cells were seeded in a 6-well plate. 6 Cells were seeded at a density of cells / well. Cells were harvested after 2, 4, and 6 days and counted using a hemocytometer. For colony formation testing, cells transformed with shControl or shGLUT3 were seeded in 6-well plates at a density of 500 cells / well for 2 weeks, fixed in 4% formalin at room temperature for 30 minutes, and stained with 0.5% crystal violet.

[0257]

[0258] <Reference Example 3> Western blot

[0259] Total protein from the cells was extracted using lysis buffer supplemented with a protease inhibitor (Roche Applied Science, #4906845001), incubated on ice for 10 minutes, and then centrifuged at 13,000 rpm for 15 minutes at 4°C to obtain the supernatant. Protein concentration was measured using a bicinchoninic acid protein assay kit (Thermo Fisher, #23225). Equal amounts of protein were separated by SDS-PAGE and transferred to a PVDF membrane (Millipore, #IPVH00010). The membrane was incubated overnight with the primary antibody at 4°C and then incubated with the secondary antibody. Protein bands were visualized using Immobilon® Forte Western HRP substrate (Millipore, #223633).

[0260]

[0261] <Reference Example 4> Immunohistochemical (IHC) Staining

[0262] Paraffin-fixed samples were sectioned to a thickness of 4 μm, the paraffin was removed, and the antigens were recovered after rehydration. Then, the sections were blocked with 3% normal goat serum and inoculated with the primary antibody anti-GLUT3 (abcam, #ab41525), diluted 500:1 in BSA, at 4°C for one day. The slides were washed five times with PBST, stained with the Vecrastain Elite ABC HRP Kit (Vector, Burlingame, #PK-6101), and then visualized using a Leica microscope (Leica Microsystems, Wetzlar, Germany, #DM750).

[0263]

[0264] <Reference Example 5> Immunofluorescence (IF) staining

[0265] For shControl or shRNA-mediated knockdown of GLUT3, cells were seeded onto chamber slides (Thermo Fisher Scientific, #154534) the night before and fixed with 4% formalin for 15 minutes. After permeabilization with 0.5% Triton X-100 at room temperature for 15 minutes, cells were blocked with 5% bovine serum albumin for 1 hour, followed by overnight incubation with the primary antibody at 4°C. Then, fluorescein isothiocyanate-conjugated secondary antibody (Invitrogen, Carlsbad, CA, USA, #A-11029) was applied for 1 hour in the dark, and nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole) for 20 minutes. IF images were captured using a confocal microscope (Leica Microsystems, #TCS SP5 II).

[0266]

[0267] <Reference Example 6> Flow cytometry analysis

[0268] 1 × 10⁶ cells in a 6-well plate 5 Cells were seeded at a cell / well density and cultured for 24 hours. Then, cells were harvested and fixed in 70% ethanol at 4°C for 1 hour. The harvested cells were added to wells containing 1 ml PBS, 50 g / ml propidium iodide (PI), and 100 g / ml RNase A, and then incubated in a dark room at room temperature for 30 minutes. Cells were detected using a CyAn ADP analyzer (Beckman Coulter, #27372).

[0269]

[0270] <Reference Example 7> Measurement of glucose absorption and internal ATP

[0271] To measure glucose uptake, place 2 × 10 cells in a 6-well plate 3 Cells were seeded at a density of cells / well and cultured for 24 hours. The next day, cells were washed twice with 1× PBS, and glucose levels were determined using the Glucose Uptake-Glo™ assay kit (Promega, #J1342) according to the manufacturer's instructions.

[0272] To detect intracellular ATP, place 5 × 10 cells in a 6-well plate 5 After dispensing at a density of cells / well and 24 hours later, relative ATP levels were measured using an ELITEN ATP assay system (Promega, #FF2000) according to the manufacturer's instructions. Intracellular ATP levels were measured via luminescence signals.

[0273]

[0274] <Reference Example 8> Mouse xenotransplantation experiment

[0275] All mouse experiments were conducted with the approval of the Animal Ethics Committee of the Laboratory Animal Center at CHA University (Reference No.: IACUC220119). Animals were handled in an accredited animal facility in accordance with institutional guidelines and regulations under conditions of 72 ± 2 °F, 35 ± 55% relative humidity, a 12-hour light-dark cycle, and the absence of specific pathogens. For cell-derived xenograft experiments, 1 × 10⁶ cells of the designated SW620-shControl or SW620-shGLUT3 stable cell line were implanted in the flanks of the mice. 6 Dogs were suspended in 100 μL of PBS and injected subcutaneously (n = 5 per group), and were sacrificed by carbon dioxide euthanasia on day 14 to avoid excessive tumor burden. The size and weight of the tumors were measured using calipers, and their volumes were calculated. All tumors were fixed overnight at room temperature and stored in 4% paraformaldehyde.

[0276]

[0277] <Reference Example 9> RNA Sequencing

[0278] RNA libraries were constructed using Lexogen's QuantSeq 3' mRNA-Seq Library Preparation Kit according to the manufacturer's instructions. Specifically, 500 ng of total RNA was hybridized with oligo-dT primers containing Illumina-compatible sequences for reverse transcription. After the RNA template was degraded, a second strand was synthesized using random primers with Illumina-compatible linker sequences. Subsequently, the double-stranded libraries were purified and amplified, and single-end sequencing was performed using NextSeq 500 (Illumina, USA). Gene classification was based on results retrieved from the DAVID and Medline databases, and data mining and graphical visualization were performed using ExDEGA (Ebiogen Inc., Korea).

[0279]

[0280] <Reference Example 10> Luciferase reporter assay

[0281] Cells were transiently transfected with the GLUT3 luciferase reporter plasmid in 6-well plates for 24 hours, and then treated with the specified compound for 6 hours. Luciferase activity was measured using a luciferase assay system (Promega, #1500) according to the manufacturer's protocol.

[0282]

[0283] Examples and Comparative Examples

[0284] <Example 1> 4-(4-(4-chlorophenethoxy)phenyl-N-(4-(2-(diethylamino)ethoxy)phenyl)-1,3,5-triazin-2-amine (4-(4-(4-chlorophenethoxy)phenyl)-N-(4-(2-(diethylamino)ethoxy)phenyl)-1,3,5-triazin-2-amine) (Compound SGT-24-104)

[0285] <Comparative Example> 4-chloro-6-(4-4-chlorophenethoxy)phenyl)-N-(4-2-(diethylamino)ethoxy)phenyl)-1,3,5-triazin-2-amine (Compound SGT-24-101)

[0286] Compound SGT-24-104 of Example 1 and Comparative Example compound SGT-24-101, which are compounds having a selective inhibitory effect on GLUT3, were synthesized as shown in Reaction Scheme 1 below (Fig. 30).

[0287]

[0288] [Reaction Equation 1]

[0289]

[0290]

[0291] Step 1: Synthesis of 1-bromo-4-(4-chlorophenethoxy)benzene (compound (5))

[0292] 4-bromophenol (5.00 g, 28.90 mmol), 4-chlorophenethyl alcohol (4.69 mL, 34.68 mmol), and triphenylphosphine (9.10 g, 34.68 mmol) were dissolved in anhydrous tetrahydrofuran (150 mL), diisopropyl azodicarboxylate (6.83 mL, 34.68 mmol) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, water was added dropwise, and the mixture was extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (ethyl acetate:n-hexane = 1:30) to obtain 1-bromo-4-(4-chlorophenethoxy)benzene in a yield of 8.74 g, or 97%.

[0293] of the above-mentioned obtained 1-bromo-4-(4-chlorophenethoxy)benzene 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0294] 1 H NMR (CDCl3, 800 MHz) δ7.34 (d,J= 8.8 Hz, 2H), 7.26 (d,J= 8.4 Hz, 2H), 7.18 (d,J= 8.4 Hz, 2H), 6.74 (d,J= 9.0 Hz, 2H), 4.09 (t,J= 6.8 Hz, 2H), 3.03 (t,J= 6.8 Hz, 2H),

[0295] 13 C NMR (CDCl3, 200 MHz) δ157.8, 136.5, 132.4, 132.3 (2C), 130.3 (2C), 128.6 (2C), 116.3 (2C), 113.0, 68.5, 35.0.

[0296]

[0297] Step 2: Synthesis of 2,4-dichloro-6-(4-(4-chlorophenethoxy)phenyl)-1,3,5-triazine (compound (6))

[0298] 1-bromo-4-(4-chlorophenethoxy)benzene (1.00 g, 3.21 mmol) prepared in Step 1 above was dissolved in anhydrous tetrahydrofuran (35 mL) and cooled to -78 °C, after which n-butyllithium (2.5 M, 1.41 mL, 3.53 mmol) was added dropwise. After stirring for 1 hour at the same temperature, cyanuric chloride (621 mg, 3.37 mmol) was added and stirred for 1 hour at room temperature. After the reaction was complete, water was added dropwise, and the mixture was extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate:n-hexane = 1:15) to obtain 2,4,-dichloro-6-(4-(4-chlorophenethoxy)phenyl)-1,3,5-triazine in a yield of 403 mg, 32%.

[0299] of the above-mentioned 2,4-dichloro-6-(4-(4-chlorophenethoxy)phenyl)-1,3,5-triazine 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0300] 1 H NMR (DMSO-d6, 800 MHz) δ8.43 (d,J= 9.0 Hz, 2H), 7.28 (d,J= 8.4 Hz, 2H), 7.21 (d,J= 8.3 Hz, 2H), 6.95 (d,J= 9.0 Hz, 2H), 4.23 (t,J= 6.8 Hz, 2H), 3.09 (t,J= 6.8 Hz, 2H),

[0301] 13C NMR (CDCl3, 200 MHz) δ174.1, 171.6 (2C), 164.3, 136.2, 132.6, 132.3 (2C), 130.3 (2C), 128.7 (2C), 125.1, 114.9 (2C), 68.7, 34.9.

[0302]

[0303] Step 3: Synthesis of 4-chloro-6-(4-4-chlorophenethoxy)phenyl)-N-(4-2-(diethylamino)ethoxy)phenyl)-1,3,5-triazin-2-amine (compound SGT-24-101)

[0304] 2,4-dichloro-6-(4-(4-chlorophenethoxy)phenyl)-1,3,5-triazine (200 mg, 0.53 mmol) prepared in Step 2 above was dissolved in anhydrous 1,4-dioxane (5 mL), and 4-(2-(diethylamino)ethoxy)aniline (0.11 mL, 0.55 mmol) was added and stirred at room temperature for 6 hours. After the reaction was complete, a saturated aqueous sodium bicarbonate solution was added dropwise, extracted three times with ethyl acetate, dried and filtered with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (dichloromethane:methanol = 1:15) to obtain 4-chloro-6-(4-4-chlorophenethoxy)phenyl)-N-(4-2-(diethylamino)ethoxy)phenyl)-1,3,5-triazine-2-amine (Formula 4) in a yield of 255 mg, 88%:

[0305] [Chemical Formula 4]

[0306]

[0307] of the above-obtained 4-chloro-6-(4-4-chlorophenethoxy)phenyl)-N-(4-2-(diethylamino)ethoxy)phenyl)-1,3,5-triazine-2-amine (compound SGT-24-101; formula 4). 1As a result of confirming the H NMR, the following data was obtained:

[0308] 1 H NMR (CDCl3, 800 MHz) δ8.33 (d,J= 8.8 Hz, 2H), 7.49 (bs, 1H), 7.46 (bs, 1H), 7.29 (bs, 1H), 7.27 (d,J= 8.4 Hz, 2H), 7.20 (d,J= 8.3 Hz, 2H), 6.92 (d,J= 8.9 Hz, 4H), 4.21 (t,J= 6.8 Hz, 2H), 4.10 (bs, 2H), 3.07 (t,J= 6.8 Hz, 2H), 2.93 (bs, 2H), 2.70 (bs, 4H), 1.10 (bs, 6H)

[0309] 13 C NMR (CDCl3, 200 MHz) δ172.8, 170.3, 164.4, 162.9, 155.9, 136.4, 132.5, 131.3 (2C), 130.9, 130.3 (2C), 128.6 (2C), 127.2, 122.8 (2C), 114.9 (2C), 114.4 (2C), 68.5, 66.5, 51.6, 47.8 (2C), 35.0, 11.5 (2C).

[0310]

[0311] Step 4: Synthesis of 4-(4-(4-chlorophenethoxy)phenyl-N-(4-(2-(diethylamino)ethoxy)phenyl)-1,3,5-triazin-2-amine) (compound SGT-24-104)

[0312] 4-chloro-6-(4-4-chlorophenethoxy)phenyl)-N-(4-2-(diethylamino)ethoxy)phenyl)-1,3,5-triazine-2-amine (120 mg, 0.22 mmol) prepared in Step 3 above was dissolved in methanol (5 mL), and after adding Raney-nickel (120 mg), the mixture was switched with hydrogen gas and stirred at room temperature for 8 hours. After the reaction was complete, the mixture was filtered through Celite and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (dichloromethane:methanol = 15:1) to obtain the final compound, 4-(4-(4-chlorophenethoxy)phenyl-N-(4-(2-(diethylamino)ethoxy)phenyl)-1,3,5-triazine-2-amine, in a yield of 70 mg, 62%.

[0313] [Chemical Formula 2]

[0314]

[0315] of the obtained 4-(4-(4-chlorophenethoxy)phenyl-N-(4-(2-(diethylamino)ethoxy)phenyl)-1,3,5-triazine-2-amine (compound SGT-24-104; formula 2) 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0316] 1 H NMR (CDCl3, 800 MHz) δ8.66 (s, 1H), 8.35 (d,J= 9.0 Hz, 2H), 7.51 (bs, 2H), 7.27 (d,J= 8.4 Hz, 2H), 7.21 (d,J= 8.5 Hz, 2H), 6.94 (d,J= 9.0 Hz, 2H), 6.92 (d,J= 8.3 Hz, 2H), 4.21 (t,J= 6.8 Hz, 2H), 4.11 (bs, 2H), 3.08 (t,J= 6.8 Hz, 2H), 2.94 (bs, 2H), 2.71 (bs, 4H), 1.11 (t,J=6.6 Hz, 6H)

[0317] 13C NMR (CDCl3, 200 MHz) δ170.9, 166.4 (2C), 163.9, 162.4, 155.3, 137.9, 131.0, 130.5, 129.0 (2C), 128.5 (2C), 128.2, 126.6, 123.2, 122.5, 114.9 (2C), 114.4 (2C), 68.8, 65.7, 51.4, 47.6 (2C), 35.6, 10.8 (2C).

[0318]

[0319] <Example 2> N-(4-(2-diethylamino)ethoxy)phenyl)-(4-(4-phenethoxyphenyl)-1,3,5-triazin-2-amine (N-(4-(2-(diethylamino)ethoxy)phenyl)-4-(4-phenethoxyphenyl)-1,3,5-triazin-2-amine) (Compound SGT-24-103)

[0320] Compound SGT-24-103 of Example 2, which is a compound having a selective inhibitory effect on GLUT3, was synthesized as shown in Reaction Scheme 2 below (Fig. 55).

[0321] [Reaction Equation 2]

[0322]

[0323]

[0324] Step 1: Synthesis of 1-bromo-4-phenethoxybenzene (compound (1))

[0325] 4-bromophenol (5.00 g, 28.90 mmol), phenethyl alcohol (4.17 mL, 34.68 mmol), and triphenylphosphine (9.10 g, 34.68 mmol) were dissolved in anhydrous tetrahydrofuran (150 mL), diisopropyl azodicarboxylate (6.83 mL, 34.68 mmol) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, water was added dropwise, and the mixture was extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (ethyl acetate:n-hexane = 1:30) to obtain 7.85 g of 1-bromo-4-phenethoxybenzene with a yield of 98%.

[0326] of the above-mentioned obtained 1-bromo-4-phenethoxybenzene 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0327] 1 H NMR (CDCl3, 800 MHz) δ7.33 (d,J= 9.1 Hz, 2H), 7.30 (t,J= 7.5 Hz, 2H), 7.25 (d,J= 7.0 Hz, 2H), 7.22 (d,J= 7.3 Hz, 1H), 6.75 (d,J= 8.9 Hz, 2H), 4.12 (t,J= 7.1 Hz, 2H), 3.07 (t,J= 7.1 Hz, 2H)

[0328] 13 C NMR (CDCl3, 200 MHz) δ157.9, 138.0, 132.2 (2C), 129.0 (2C), 128.5 (2C), 126.6, 116.3 (2C), 112.9, 68.9, 35.7.

[0329] LR-MS (FAB + ) m / z 276 (M + ); HR-MS (FAB + ) calcd for C 14 H 13 BrO (M +) 276.0150, found 276.0140.

[0330]

[0331] Step 2: Synthesis of 2,4-dichloro-6-(4-phenethoxyphenyl)-1,3,5-triazine (compound (2))

[0332] 1-bromo-4-phenethoxybenzene (1.50 g, 5.41 mmol) prepared in Step 1 above was dissolved in anhydrous tetrahydrofuran (55 mL) and cooled to -78 °C, after which n-butyllithium (2.5 M, 2.60 mL, 6.49 mmol) was added dropwise. After stirring for 1 hour at the same temperature, cyanuric chloride (1.30 g, 7.04 mmol) was added and stirred for 1 hour at room temperature. After the reaction was complete, water was added dropwise, and the mixture was extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate:n-hexane = 1:30) to obtain 899 mg of 2,4-dichloro-6-(4-phenethoxyphenyl)-1,3,5-triazine in a yield of 48%.

[0333] of the above-mentioned 2,4,-dichloro-6-(4-phenethoxy)phenyl)-1,3,5-triazine 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0334] 1 H NMR (CDCl3, 800 MHz) δ8.43 (d,J= 9.1 Hz, 2H), 7.32 (t,J= 7.5 Hz, 2H), 7.28 (d,J= 6.7 Hz, 2H), 7.25 (d,J= 7.3 Hz, 1H), 6.97 (d,J= 8.9 Hz, 2H), 4.26 (t,J= 7.1 Hz, 2H), 3.13 (t,J= 7.1 Hz, 2H)

[0335] 13 C NMR (CDCl3, 200 MHz) δ174.1, 171.6 (2C), 164.4, 137.6, 132.3 (2C), 129.0 (2C), 128.6 (2C), 126.7, 125.0, 114.9 (2C), 69.1, 35.6.

[0336] LR-MS (FAB + ) m / z 346 (M+H + ); HR-MS (FAB + ) calcd for C 17 H 14 Cl2N3O (M+H + ) 346.0514, found 346.0506.

[0337]

[0338] Step 3: Synthesis of 4-chloro-N-(4-(2-(diethylamino)ethoxy)phenyl)-6-(4-phenethoxy)phenyl)-1,3,5-triazin-2-amine (compound (3)).

[0339] 2,4,-dichloro-6-(4-phenethoxy)phenyl)-1,3,5-triazine (220 mg, 0.64 mmol) prepared in Step 2 above was dissolved in anhydrous 1,4-dioxane (6 mL), and 4-(2-(diethylamino)ethoxy)aniline (132 mg, 0.64 mmol) was added and stirred at room temperature for 6 hours. After the reaction was complete, a saturated aqueous sodium bicarbonate solution was added dropwise, extracted three times with ethyl acetate, dried and filtered with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (dichloromethane:methanol = 20:1) to obtain 4-chloro-N-(4-(2-(diethylamino)ethoxy)phenyl)-6-(4-phenethoxy)phenyl)-1,3,5-triazine-2-amine in a yield of 303 mg, 92%:

[0340] of the above-obtained 4-chloro-N-(4-(2-(diethylamino)ethoxy)phenyl)-6-(4-phenethoxy)phenyl)-1,3,5-triazine-2-amine 1 As a result of confirming the H NMR, the following data was obtained:

[0341] 1 H NMR (CDCl3, 800 MHz) δ8.31 (d,J= 8.5 Hz, 2H), 7.58 (d,J= 6.1 Hz, 1H), 7.50 (bs, 1H), 7.30 (t,J= 7.5 Hz, 2H), 7.27 (d,J= 7.2 Hz, 2H), 7.23 (t,J= 7.4 Hz, 2H), 6.92 (d,J= 8.6 Hz, 2H), 6.86 (bs, 2H), 4.54 (bs, 2H), 4.22 (t,J= 7.1 Hz, 2H), 3.76 (bs, 2H), 3.47 (bs, 4H), 3.10 (t,J= 7.1 Hz, 2H), 1.37 (t,J= 7.1 Hz, 6H)

[0342] 13 C NMR (CDCl3, 200 MHz) δ176.0. 172.6, 170.2, 164.3, 162.9, 154.4, 137.8, 131.2 (2C), 129.0 (2C), 128.5 (2C), 126.6 (2C), 122.9 (2C), 114.6 (2C), 114.4 (2C), 68.8, 63.6, 61.9, 60.2 (2C), 35.6, 8.7 (2C)

[0343] LR-MS (FAB + ) m / z 518 (M+H + ); HR-MS (FAB + ) calcd for C 29 H 33 ClN5O2(M+H + ) 518.2323, found 518.2336.

[0344]

[0345] Step 4: Synthesis of N-(4-(2-diethylamino)ethoxy)phenyl)-(4-(4-phenethoxyphenyl)-1,3,5-triazin-2-amine) (SGT-24-103)

[0346] 4-chloro-N-(4-(2-(diethylamino)ethoxy)phenyl)-6-(4-phenethoxy)phenyl)-1,3,5-triazine-2-amine (100 mg, 0.19 mmol) prepared in Step 3 above was dissolved in ethyl acetate:methanol (1:1, 4 mL), palladium (hydroxide) 2 (15 mg) was added, and the mixture was replaced with hydrogen gas and stirred at room temperature for 6 hours. After the reaction was complete, the mixture was filtered through Celite and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (dichloromethane:methanol = 20:1) to obtain the final candidate compound, N-(4-(2-diethylamino)ethoxy)phenyl)-(4-(4-phenethoxyphenyl)-1,3,5-triazine-2-amine (Formula 3), in a yield of 70 mg, 75%:

[0347] [Chemical Formula 3]

[0348]

[0349] of the above-obtained N-(4-(2-diethylamino)ethoxy)phenyl)-(4-(4-phenethoxyphenyl)-1,3,5-triazine-2-amine 1 H NMR and 13 As a result of confirming the C NMR, the following data was obtained:

[0350] 1H NMR (CDCl3, 800 MHz) δ8.66 (s, 1H), 8.35 (d,J= 9.0 Hz, 2H), 7.52 (bs, 2H), 7.32-7.30 (m, 2H), 7.28 (d,J= 6.7 Hz, 2H), 7.23 (d,J= 7.2 Hz, 1H), 6.96 (d,J= 8.9 Hz, 2H), 6.92 (d,J= 8.4 Hz, 2H), 4.24 (t,J= 7.1 Hz, 2H), 4.20 (bs, 2H), 3.12 (t,J= 7.1 Hz, 2H), 3.04 (bs, 2H), 2.82 (bs, 4H), 1.18 (bs, 6H)

[0351] 13 C NMR (CDCl3, 200 MHz) δ170.9, 166.4 (2C), 163.9, 162.4, 155.3, 137.9, 131.0, 130.5, 129.0 (2C), 128.5 (2C), 128.2, 126.6, 123.2, 122.5, 114.9 (2C), 114.4 (2C), 68.8, 65.7, 51.4, 47.6 (2C), 35.6, 10.8 (2C).

[0352] LR-MS (FAB + ) m / z 484 (M+H + ); HR-MS (FAB + ) calcd for C 29 H 34 N5O2(M+H + ) 484.2713, found 484.2707.

[0353]

[0354] Experimental Example

[0355] <Experimental Example 1> Analysis of the association between GLUT3 and colorectal cancer

[0356] (1) Analysis of GLUT3 expression levels in colorectal cancer patients

[0357] To confirm the role of GLUT3 in the progression of colorectal cancer, analyses such as mRNA, IHC, and Western blot were performed on tissues from colorectal cancer patients.

[0358] All human colon tissues used in the experiment (90 pairs of cancer cell and normal cell samples) were collected from CHA University Bundang Hospital, and the clinical specimens were used with the approval of the Ethics Committee of CHA University Bundang Hospital (1044308-202103-BR-015-02).

[0359] As a result of analyzing the publicly available bioinformatics database, it was confirmed that GLUT3 is involved in intracellular metabolic processes and contributes to energy production and metabolic homeostasis, as shown in Figure 1.

[0360] Accordingly, changes in GLUT3 expression levels were confirmed to have upregulated GLUT3 mRNA in colorectal cancer tissue compared to normal tissue (Fig. 2). Immunohistochemistry (IHC) analysis also confirmed that, consistent with the mRNA results, only minimal GLUT3 expression was observed in normal tissue, while GLUT3 expression was significantly higher in colorectal cancer tissue (Fig. 3). Furthermore, Western blot analysis similarly confirmed that GLUT3 protein was upregulated in colorectal cancer tissue compared to normal tissue (Fig. 4).

[0361] Additionally, as a result of performing Kaplan-Meier survival analysis using the TCGA dataset, it was confirmed that, as shown in Figure 5, colorectal cancer patients with high GLUT3 mRNA expression levels showed significantly lower overall survival rates compared to those with low GLUT3 expression levels.

[0362] In addition, a comprehensive evaluation of all GLUT isoforms in colorectal cancer tissue using RNA-seq data revealed significant changes in the expression of 12 GLUT isoforms, with GLUT1 and GLUT3 showing the most pronounced upregulation in colorectal cancer tissue (Fig. 6).

[0363] When the above results were combined, it was confirmed that GLUT3 expression significantly increases during the formation and progression of colorectal cancer tumors, and accordingly, that the regulation of GLUT3 expression can be utilized in the treatment of colorectal cancer.

[0364]

[0365] (2) Analysis of the effects of GLUT3 on cancer cell metabolism and growth

[0366] To differentiate the functional role of GLUT3 in colorectal cancer cells, experiments were conducted using an shRNA construct targeting GLUT3 (shGLUT3) and a negative control construct (shControl) in SW620 cells characterized by high endogenous GLUT3 expression.

[0367] As shown in Fig. 7, immunofluorescence (IF) analysis confirmed that GLUT3 protein levels were significantly reduced when shGLUT3 was treated compared to shControl. Additionally, when GLUT3 was silenced by treating SW620 cells with shGLUT3, cell proliferation and growth were inhibited compared to shControl, resulting in a significant decrease in cell viability (Fig. 8, left), while cell proliferation was increased when GLUT3 was ectopically overexpressed (Fig. 8, right).

[0368] Consistent with the results of Figures 7 and 8, the results of colony formation and anchorage-independent growth assays showed that the colony formation and invasive potential of cells treated with shGLUT3 were significantly reduced compared to shControl (Figures 9 and 10). Additionally, flow cytometry was performed to specifically evaluate the effect of GLUT3 on the cell cycle, and as a result, as shown in Figure 11, significant arrest of the G1 / S phase was induced in SW620 cells when treated with shGLUT3.

[0369] In addition, considering that glucose metabolism plays an important role in cell growth, the effect of GLUT3 regulation on glycolytic activity in colorectal cancer cells was evaluated. When shGLUT3 was treated, glucose uptake and ATP production decreased in SW620 cells compared to shControl (Figs. 12 and 13), but when GLUT3 was ectopically overexpressed, glucose uptake and ATP production increased (Figs. 14 and 15).

[0370] Specifically, when evaluating the effect of GLUT3 on tumor growth in a subcutaneous xenograft model, tumors derived from SW620 cells treated with shGLUT3 showed a 5-fold reduction in tumor growth compared to SW620 cell-derived tumors treated with shControl (Fig. 16). In addition, Gene Set Enrichment Analysis (GSEA) results on RNA-seq data from colorectal cancer patient samples with increased GLUT3 expression showed a positive correlation between high GLUT3 levels and the activation of the cell cycle and collagen metabolic pathways (Fig. 17).

[0371] Through the above results, it was confirmed that GLUT3 has potential as a therapeutic target for colorectal cancer because it promotes the rapid proliferation and invasive behavior of colorectal cancer cells.

[0372]

[0373] (3) Analysis of upstream mechanisms of GLUT3 regulation

[0374] To identify the upstream mechanism regulating GLUT3 expression, the correlation between upstream receptor tyrosine kinases (RTKs) and GLUT3 expression was analyzed.

[0375] As a result, as shown in Figures 18 and 19, there is a significant positive correlation between IGF1 mRNA and GLUT3 mRNA expression, and the difference in GLUT3 expression was most pronounced under conditions where IGF1 expression was increased.

[0376] To further verify the role of IGF1 in GLUT3 regulation, immunohistochemistry (IHC) was performed on tissue samples from colorectal cancer patients, and it was confirmed that GLUT3 expression increased in patients with high IGF1 expression levels (Fig. 20). RNA sequencing (RNA-Seq) analysis also showed a similar pattern of increased GLUT3 expression in colorectal cancer patients with high IGF1 expression levels, whereas colorectal cancer patients with low IGF1 expression levels tended to have low GLUT3 expression (Fig. 21).

[0377] To evaluate the effect of IGF1-mediated GLUT3 regulation on survival in colorectal cancer patients, a Kaplan-Meier survival analysis was performed using the TCGA dataset. As shown in Figure 22, patients with high expression of IGF1 and GLUT3 showed significantly lower overall survival rates compared to patients with low IGF1 expression.

[0378] Additionally, GLUT3 protein levels (Fig. 23) and glucose metabolism (Fig. 24) were evaluated in HCT116 cells after knocking out IGF1R. As a result, it was confirmed that GLUT3 expression decreased in the case of knocking out IGF1R (si-IGF1R) compared to the control group without knockout (si-Control) (Fig. 23), and glucose uptake and ATP production in HCT116 cells decreased (Fig. 24).

[0379] When the above results were combined, it was confirmed that there is a significant positive correlation between IGF1 and GLUT3, and that compounds capable of regulating IGF1-mediated GLUT3 expression can be used as treatments for colorectal cancer.

[0380]

[0381] <Experimental Example 2> Screening of GLUT3 Inhibitors

[0382] To identify GLUT3 selective inhibitors, compounds were treated to HCT116 cells, and then cell growth inhibition, GLUT3 protein expression levels, glucose metabolism, and cytotoxicity to normal cells were evaluated.

[0383] As a result of treating HCT116 cells with each compound for 24 hours and evaluating cell growth, in-house library compounds based on skeletal structures such as benzopyran, flavonoid, triterpenoid, steroid, catechol, isoxazole, isoxazoline, triazole, tetrazole, pyrazinone, pyranoside, pyrimidine, quinolone, quinazoline, quinazolinone, benzothiazole, benzoxazole, and benzimidazole For 22 out of 100 compounds, cell growth was significantly reduced compared to the control group that was not treated with the compound (Fig. 25).

[0384] Accordingly, the top 8 of the 22 compounds capable of acting as GLUT3 inhibitors were selected, and additional Western blots were performed. As shown in Fig. 26, when treated with the 93rd hit compound (SGT-24093), GLUT3 protein levels were most significantly downregulated after 24 hours, confirming that SGT-24093 has the potential to be a potent inhibitor of GLUT3. In addition, the effects of compound treatment on glucose uptake and ATP levels in HCT116 cells were evaluated, and it was confirmed that SGT-24093 most significantly reduced glucose uptake (Fig. 27) and ATP production (Fig. 28) in HCT116 colorectal cancer cells.

[0385] To evaluate the selective cancer cell treatment and safety of SGT-24093, cytotoxicity was compared in normal colon epithelial cells (IEC6) and HCT116 cancer cells. It was confirmed that SGT-24093 did not cause toxicity in normal cells but selectively inhibited the proliferation of cancer cells (Fig. 29).

[0386] Through the above results, it was confirmed that SGT-24093, which inhibits GLUT3 expression upregulated in colorectal cancer patients, inhibits glucose metabolism related to cancer cell growth, is non-toxic to normal cells, and specifically inhibits only cancer cells, can be utilized as an effective GLUT3 inhibitor, that is, a treatment for colorectal cancer.

[0387]

[0388] <Experimental Example 3> Evaluation of the Efficacy of SGT-24093 as a GLUT3 Inhibitor

[0389] In order to evaluate whether not only the 93rd hit compound (SGT-24093) but also SGT-24-101, a compound derived during the synthesis process of the said compound, has efficacy as a GLUT3 inhibitor, experiments were conducted to evaluate the cytotoxicity on normal cells and the effects on glucose metabolism.

[0390] As shown in Figure 31, both SGT-24093 and SGT-24-101 were confirmed to be safe as they did not exhibit cytotoxicity in IEC6. However, when HCT116 cancer cells were treated with SGT-24093, cell viability was significantly lower, whereas when treated with SGT-24-101, no significant decrease was observed, confirming that SGT-24093 exhibited superior efficacy.

[0391] In addition, glucose uptake and ATP production, which are key indicators of cancer cell metabolism, were measured. When treated with SGT-24093, both glucose uptake (Fig. 32) and ATP production (Fig. 33) were significantly reduced, whereas when treated with SGT-24-101, no significant difference was observed compared to the control group that was not treated with the compound.

[0392] Based on the above results, treatment with SGT-24093 showed a significantly superior inhibitory effect on the GLUT3-mediated metabolic pathway of cancer cells compared to compound SGT-24-101, which is attributed to structural modifications during the synthesis process, confirming that SGT-24093 can be utilized as an effective GLUT3 inhibitor.

[0393]

[0394] <Experimental Example 4> Evaluation of the Mechanism of Action of SGT-24093 as a GLUT3 Inhibitor

[0395] To confirm the selectivity and mechanism of action of compound 93 (SGT-24093) as a GLUT3 inhibitor, selective inhibition of cancer cell proliferation and selective GLUT3 action were evaluated.

[0396] As shown in Fig. 34, SGT-24093 did not affect the viability of normal colon epithelial cells and selectively inhibited only the growth of HCT116 cancer cells. In addition, the IC of SGT-24093 in HCT116 cells 50 The result of measuring was found to be 10.49 μM (Fig. 35).

[0397] Next, the GLUT3 promoter activity was evaluated to confirm the direct effect of the compound on GLUT3 expression. As shown in Figure 36, it was confirmed that when SGT-24093 was treated, GLUT3 promoter activity was significantly reduced, and GLUT3 transcription was strongly inhibited.

[0398] To confirm the function of SGT-24093 as a selective inhibitor of GLUT3, its action on GLUT1 and GLUT4, which have structural homology with GLUT3 and are members of the class I glucose transporter family, was also measured. As a result, it was confirmed that SGT-24093 selectively inhibited only GLUT3 expression without affecting the protein levels of GLUT1 or GLUT4 (Fig. 37).

[0399] In addition, for further functional analysis, the effect on glucose metabolism was evaluated, and it was confirmed that SGT-24093 significantly reduced glucose uptake and ATP production, thereby inhibiting the metabolism of cancer cells (Fig. 38).

[0400] In addition, when it was determined whether SGT-24093 inhibits GLUT3 expression by inhibiting IGF1R signaling, as shown in Fig. 39, treatment with compound SGT-24093 significantly reduced the phosphorylation level of IGF1R and decreased the expression of GLUT3 protein, confirming that SGT-24093 downregulated GLUT3 by inhibiting IGF1R phosphorylation. Furthermore, treatment with lincitinib, known as an IGF1R inhibitor, also reduced GLUT3 expression, similar to the case with SGT-24093, confirming the role of IGF1R signaling in GLUT3 regulation (Fig. 40).

[0401] In other words, it was confirmed that SGT-24093 selectively inhibits GLUT3 without affecting structurally similar glucose transporters GLUT1 and GLUT4, and that this is mediated through the inhibition of IGF1R phosphorylation.

[0402]

[0403] <Experimental Example 5> Evaluation of the Effect of Combination Administration of SGT-24093 and Lincitinib

[0404] To determine whether the cancer cell treatment effect is excellent when the 93rd hit compound (SGT-24093) and lincitinib are administered in combination at varying concentrations (0, 0.1, 1, 10, and 100 μM), the cancer cell survival inhibition ability upon combination administration was evaluated.

[0405] As a result of evaluating the combined effect of SGT-24093 and lincitinib on cancer cell survival, as shown in Figure 41, it was confirmed that a synergistic effect exists, as simultaneous treatment of cells with both compounds exhibited significantly superior inhibition of cancer cell survival compared to treatment with each compound alone. Specifically, when lincitinib and SGT-24093 were treated alone, SGT-24093 showed a lower cancer cell survival rate than lincitinib. Similarly, when treated in combination, for example, the decrease in cancer cell survival rate was higher when lincitinib 0.1 μM and SGT-24093 100 μM were treated compared to when lincitinib 100 μM and SGT-24093 0.1 μM were treated. Through this, it was confirmed that SGT-24093 exhibits a significantly superior inhibitory effect on cancer cell proliferation even when administered in combination.

[0406] In other words, SGT-24093 exhibited excellent cancer cell inhibitory ability when combined with lincitinib, confirming that it is promising as a targeted therapy for GLUT3-dependent cancers not only when used alone but also when administered in combination.

[0407]

[0408] <Experimental Example 6> Further screening and efficacy evaluation of SGT-24-103, a GLUT 3 inhibitor

[0409] In order to select compounds that exhibit an effect equivalent to or superior to hit compound No. 93 (SGT-24093), which was selected as a GLUT3 selective inhibitor in Experimental Examples 2 to 5 above, the same process as in Experimental Example 2 was performed by adding types of compounds (101, 102, SGT-24-103, 105, 106, etc.). Specifically, after treating HCT116 cells with each compound, cell growth inhibition, GLUT3 protein expression levels, glucose metabolism, and cytotoxicity to normal cells were evaluated.

[0410] As a result of treating HCT116 cells with each compound for 24 hours and evaluating cell growth, in-house library compounds based on skeletal structures such as benzopyran, flavonoid, triterpenoid, steroid, catechol, isoxazole, isoxazoline, triazole, tetrazole, pyrazinone, pyranoside, pyrimidine, quinolone, quinazoline, quinazolinone, benzothiazole, benzoxazole, and benzimidazole Significant inhibition of cell growth compared to the control group was confirmed in 29 of the 105 compounds (Fig. 42), and 10 of them, corresponding to the top 30%, were selected.

[0411] Among the 10 selected compounds, toxicity to normal colon cells was verified first, and as a result, it was confirmed that compound SGT-24-103 did not cause toxicity to normal cells and selectively inhibited the proliferation of cancer cells (Figs. 43, 44).

[0412] Next, when the inhibitory effect of compound SGT-24-103 on colon cancer cells was compared with other compounds, it was confirmed that not only was the cancer cell inhibitory activity the best (Fig. 44), but glucose metabolism and ATP production were also effectively inhibited compared to other compounds (Figs. 45, 46).

[0413] In addition, the IC of SGT-24-103 against various colorectal cancer cell lines 50 As a result of measuring the values, concentration-dependent inhibitory activity on cancer cells was observed (Fig. 47), glucose metabolism and ATP production were also inhibited in a concentration-dependent manner (Fig. 48), and it was confirmed that the inhibitory effect on the colony formation ability of cancer cells was also significant (Fig. 49).

[0414] Meanwhile, to determine whether compound SGT-24-103 could selectively inhibit only GLUT3, the inhibitory effect on other isoforms was compared, and as a result, it was confirmed that it could inhibit only GLUT3 without inhibiting other isoforms (GLUT1, GLUT4) other than GLUT3 (Fig. 50).

[0415] In addition, the inhibitory activity of compound SGT-24-103 against various cancers was evaluated, and it was confirmed that it has an inhibitory effect on cancer cell growth in gastric cancer, liver cancer, lung cancer, breast cancer, prostate cancer, and glioblastoma cell lines (Figs. 51 to 54).

[0416] Through the above results, it was confirmed that compound SGT-24-103 can be utilized as a treatment for colorectal cancer as an effective GLUT3 inhibitor that inhibits GLUT3 expression upregulated in colorectal cancer patients, inhibits glucose metabolism associated with cancer cell growth, is non-toxic to normal cells, and specifically inhibits only cancer cells.

Claims

1. A compound represented by the following chemical formula 1 or a salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

2. In Claim 1, The above R 1 and R 2 Each is independently H, F, Cl, Br, or I; The above R 3 and R 4 A compound or a salt thereof, each independently being a C1 to C4 alkyl.

3. In Claim 1, The above compound is a compound or a salt thereof, wherein the compound is represented by the following chemical formula 2 or chemical formula 3: [Chemical Formula 2] , [Chemical Formula 3] .

4. A pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

5. A pharmaceutical composition according to claim 4, wherein the cancer is one in which the expression or activity of the GLUT (glucose transporter) gene or protein within the cancer cell is increased.

6. A pharmaceutical composition according to claim 5, wherein the GLUT is GLUT3.

7. A pharmaceutical composition according to claim 4, wherein the cancer is one or more selected from the group consisting of gastric cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and brain tumor.

8. A pharmaceutical composition according to claim 4, wherein the compound or a pharmaceutically acceptable salt thereof inhibits the expression or activity of the GLUT3 gene or protein.

9. A pharmaceutical composition according to claim 4, wherein the compound or a pharmaceutically acceptable salt thereof inhibits the interaction between IGF1 (insulin-like growth factor 1) and IGF1R (insulin-like growth factor 1 receptor).

10. A pharmaceutical composition according to claim 9, wherein the inhibition of the interaction between IGF1 and IGF1R is by one or more selected from the group consisting of inhibition of the expression or activity of the IGF1 gene or protein and inhibition of the expression or activity of the IGF1R gene or protein.

11. A pharmaceutical composition according to claim 10, wherein the inhibition of activity of the IGF1R protein is the inhibition of phosphorylation of the IGF1R protein.

12. A pharmaceutical composition according to claim 4, wherein the compound or a pharmaceutically acceptable salt thereof inhibits glycolysis of cancer cells.

13. A pharmaceutical composition according to claim 12, wherein the inhibition of the corresponding process is one or more selected from the group consisting of inhibition of glucose absorption within cancer cells and inhibition of ATP production within cancer cells.

14. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition further comprises an IGF1R inhibitor.

15. A pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is administered in combination with an IGF1R inhibitor.

16. A pharmaceutical composition according to claim 15, wherein the pharmaceutical composition and the IGF1R inhibitor are administered simultaneously.

17. A health functional food for the prevention or improvement of cancer comprising a compound represented by the following chemical formula 1 or a food-grade acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 to R 4 Each is independently H, F, Cl, Br, I, or a substituted or unsubstituted C1 to C6 alkyl.

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