KR20230004995A - New compounds for differentiating human tooth stem cells into odontoblasts and medical uses thereof - Google Patents

Abstract

The present invention relates to a novel compound for differentiating human dental stem cells into odontoblasts, and medical uses thereof. By confirming an effect of the novel compound on the period of differentiation of dental stem cells into odontoblasts, it is confirmed that the differentiation of odontoblasts is induced and promoted as the concentration of the novel compound increases. Accordingly, the compound can be used in therapeutic fields such as tooth or bone regeneration.

Description

New compounds for differentiating human tooth stem cells into odontoblasts and medical uses thereof {New compounds for differentiating human tooth stem cells into odontoblasts and medical uses thereof}

The present invention relates to a novel compound for differentiating human dental stem cells into odontoblasts and a medical use thereof.

Adult stem cells are stem cells that are distinct from embryonic stem cells, and have multipotent properties capable of differentiating into certain cells. In particular, in the case of dental stem cells, ectodermal mesoderm cells have an advantage in that stem cells can be obtained by easily accessing tissues compared to other tissues. Stem cells that can be classified from teeth include dental pulp stem cells (DPSC), stem cells from human exfoliated deciduous teeth (SHED), and stem cells from dental pulp. the apical part of the papilla (SCAP), stem cells from the dental follicle (DFPC), and periodontal ligament stem cells (PDLSC).

Dental stem cells suggest various possibilities as part of a method for preserving natural teeth. The entry into an aging society always pays attention to the need for research on the preservation of natural teeth in the elderly population, which is presented as a basic condition for the extension of average life expectancy. It is a trend that is taking place. Under this background, the development of a method capable of inducing or promoting the differentiation of odontoblasts from dental stem cells has been urgently required, but is still insufficient.

1. Republic of Korea Patent No. 10-1904767.

An object of the present invention is to provide a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

Another object of the present invention is a composition for differentiating odontoblasts from dental stem cells containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutical composition for tooth or bone regeneration, and a health function for tooth or bone regeneration. It is to provide food, health functional food for preservation or improvement of natural teeth, and quasi-drugs for preservation or improvement of natural teeth.

Another object of the present invention is to provide a method for differentiating dental stem cells into odontoblasts, comprising the step of inducing differentiation into odontoblasts by treating dental stem cells with the compound or a pharmaceutically acceptable salt thereof. is in

Another object of the present invention is to provide a method for preparing the compound.

In order to achieve the above object, the present invention provides a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

In addition, the present invention provides a composition for differentiating odontoblasts from dental stem cells containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutical composition for tooth or bone regeneration, and a health functional food for tooth or bone regeneration. , Health functional foods for preservation or improvement of natural teeth and quasi-drugs for preservation or improvement of natural teeth are provided.

In addition, the present invention provides a method for differentiating dental stem cells into odontoblasts, comprising treating dental stem cells with the compound or a pharmaceutically acceptable salt thereof to induce differentiation into odontoblasts.

In addition, the present invention provides a method for preparing the compound.

The present invention relates to a novel compound for differentiating human dental stem cells into odontoblasts (hereinafter referred to as JD005) and its medical use. It was found that the differentiation of odontoblasts was increased and promoted whenever the concentration of JD005 increased, and thus, a pharmaceutical composition for tooth or bone regeneration containing the same, a health functional food for tooth or bone regeneration, and a health functional food for preserving or improving natural teeth And it can be usefully used as a quasi-drug for preserving or improving natural teeth.

1 shows a method for synthesizing the novel compound JD005 for differentiating human dental stem cells into odontoblasts.
Figure 2 shows comparative experimental examples for determining the degree of differentiation of odontoblasts according to the concentration of JD005.
Figure 3 shows the expression of odontoblast differentiation markers for the group treated with JD005 and the comparison of protein expression according to the concentration of the compound treated.

Hereinafter, the present invention will be described in more detail.

Pulp-derived stem cells are mesenchymal stem cells that, like other types of stem cells, self-renew and have the characteristics of being able to differentiate into various cells in a given environment. This characteristic of differentiation induces the regeneration of dental hard tissue, which is difficult to heal naturally, and this result can play an important role in preserving natural teeth. As a result of research efforts to develop a material that can be used, a novel compound, JD005, was discovered and the present invention was completed.

The present invention provides a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

The compound is characterized in that an aminomethyl-4-phenol functional group is introduced into a triazine compound as an amino furanyl triazole.

The present invention provides a composition for differentiating odontoblasts from dental stem cells containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The dental stem cells include dental pulp stem cells (DPSC), stem cells from human exfoliated deciduous teeth (SHED), and stem cells from the apical part of the papilla (SCAP). , may be at least one selected from the group consisting of stem cells from the dental follicle (DFPC) and periodontal ligament stem cells (PDLSC), but is not limited thereto.

The dental stem cells may be separable from the dental pulp.

The composition may phosphorylate any one or more of SMAD 1, SMAD 5 and SMAD 8.

The present invention provides a method for differentiating dental stem cells into odontoblasts, comprising treating dental stem cells with the compound or a pharmaceutically acceptable salt thereof to induce differentiation into odontoblasts.

The present invention provides a pharmaceutical composition for tooth or bone regeneration containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

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

In the present invention, the pharmaceutical composition may be any one formulation selected from the group consisting of injections, granules, powders, tablets, pills, capsules, gels, suspensions, emulsions, drops or liquids according to conventional methods. .

In another embodiment of the present invention, the pharmaceutical composition is a suitable carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricant, lubricant, flavoring agent, antioxidant, buffer, bacteriostatic agent commonly used in the preparation of pharmaceutical compositions , Diluents, dispersants, surfactants, binders and may further include one or more additives selected from the group consisting of lubricants.

Specifically, carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil may be used, and solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules. These solid preparations may be prepared by mixing at least one or more excipients, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions for oral use, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and the like. 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.

According to one embodiment of the present invention, the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or It can be administered to a subject in a conventional manner via the intradermal route.

The present invention provides a health functional food for tooth or bone regeneration containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food for preserving or improving natural teeth containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The health functional food includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, It may contain organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like.

In addition, it may contain fruit flesh for the production of natural fruit juice, synthetic fruit juice and vegetable beverages. These components may be used independently or in combination. In addition, the health functional food composition is any one form of meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol and vitamin complex can be

In addition, the health functional food may additionally contain food additives, and the suitability as a "food additive" is determined according to the general rules of the Food Additive Code and general test methods approved by the Korea Food and Drug Administration unless otherwise specified. It is judged according to the relevant standards and standards.

Examples of items listed in the “Food Additives Codex” include, for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goreng pigment, guar gum, L -Mixed preparations such as sodium glutamate preparations, noodle-added alkali preparations, preservative preparations, tar color preparations, and the like.

The present invention provides a quasi-drug for preserving or improving natural teeth containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The quasi-drug may be one or more formulations selected from the group consisting of toothpaste, mouthwash, mouthwash, gum, candy, oral spray, oral ointment, oral varnish, mouthwash, and gum massage cream, but is not limited thereto. .

The present invention is a first step of mixing 2-Furoic hydrazide and S-methylisothiouronium sulfate to obtain a precipitated material by stirring in a basic aqueous solution; Water was added to the N“-(Furan-2-ylcarbonyl)carbonohydrazonic diamide obtained in the first step and stirred under microwave conditions. Then, a second step of obtaining a material by extracting and drying the reaction mixture in an organic solvent and then concentrating under reduced pressure; 5-(Furan-2-yl)-1H-1,2,4-triazol-3-amine (5-(Furan-2-yl)-1H-1,2,4-triazol obtained in the second step -3-amine) was mixed with S,S'-dimethyl N-cyanodithioiminocarbonate (S,S'-dimethyl N-cyanodithioiminocarbonate), stirred in an inert gas atmosphere, and then put into an organic solvent to extract the substance. 3rd step to obtain; 2-(furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazine-7 obtained in the third step m- to -amine (2-(Furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) A fourth step of obtaining a substance by mixing with m-chloroperoxybenzoic acid under organic solvent conditions and then stirring; and 2-(furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazine obtained in the fourth step. -7-amine (2-(Furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) A fifth step of mixing diisopropylethylamine (DIPEA) and 4-hydroxybenzylamine and then stirring to obtain a material;

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example One : odontoblast Synthesis of JD005, a novel compound with differentiation-inducing efficacy

In order to synthesize JD005 having the efficacy of inducing differentiation from dental stem cells to odontoblasts, it was synthesized in the following order.

( Step 1 ) N“-( Furan -2 - ylcarbonyl ) Carbonohydrazonic Preparation of diamide ( N ''-(Furan-2-ylcarbonyl)carbonohydrazonic diamide)

A mixture of 2-Furoic hydrazide (894 mg, 7.09 mmol) and S-methylisothiouronium sulfate (2.37 g, 8.51 mmol) was dissolved in water (12 mL). ) was added with sodium hydroxide solution (567 mg, 14.2 mmol) dissolved in it, and stirred at room temperature for 2 days. The resulting precipitated material was filtered and then washed with dimethyl ether to obtain a product as a white powder (1.02 g, yield 85.6%).

( step 2 ) 5 - ( furan -2-day)-1H-1,2,4- triazole -3- amine (5-( Furan -2- yl )-1H-1,2,4-triazol-3-amine) manufacturing

A suspension of N“-(Furan-2-ylcarbonyl)carbonohydrazonic diamide (1.02 g, 6.07 mmol) in water (20 mL) is The mixture was stirred for 1 hour under microwave conditions at 160 °C and 6 bar. The reaction mixture was extracted with ethyl acetate, washed with brine, and dried over sodium sulfate. And reduced pressure and concentration to obtain a beige powder product (815 mg, yield 89.5%) (1H-NMR (400 MHz, DMSO-D6) δ 12.11 (s, 1H), 7.71 (s, 1H), 6.71 (s, 1H), 6.58 (s, 1H), 6.10 (s, 2H)).

( Step 3 ) 2-( furan -2-day)-5-( methylthio )- [1,2,4]triazolo[1,5-a] [1,3, 5] to Liazine-7-amine (2-( Furan -2- yl )-5-( methylthio )- [1,2,4]triazolo[1,5-a] [1,3,5]triazin-7-amine) manufacturing

5-(Furan-2-yl)-1H-1,2,4-triazol-3-amine (5-(Furan-2-yl)-1H-1,2,4-triazol-3-amine) ( A mixture of 770 mg, 5.13 mmol) and S,S'-dimethyl N-cyanodithioiminocarbonate (825 mg, 5.64 mmol) was heated at 170 °C in an argon-purged atmosphere. was stirred for 3 hours. After cooling the reaction mixture to room temperature, methanol (5 mL) and dichloromethane (5 mL) were added. The reaction mixture was then refluxed overnight. The product was purified by column chromatography to obtain the product as a white solid (273 mg, yield 21.4 %).

( Step 4 ) 2-( furan -2-day)-5-( methylsulfonyl )- [1,2,4]triazolo[1,5-a] [1,3, 5] triazine -7-amine (2-( Furan -2- yl )-5-( methylsulfonyl )- [1,2,4]triazolo[1,5-a] [1,3,5]triazin-7-amine) manufacturing

22-(furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazine-7-amine (2-( Furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) (273 mg, 1.10 mmol) in dichloro The suspension with methane (8 mL) was stirred at 0 °C and a solution of m-chloroperoxybenzoic acid (<77%, 759 mg, <3.39 mmol) in dichloromethane (2 mL) was obtained. put in slowly The reaction mixture was stirred overnight at room temperature. After the solvent was removed under reduced pressure, ethanol was added. The resulting solid was filtered and then washed with ethanol to obtain an ivory powder product (248 mg, yield 80.5%).

( Step 5 ) 4-(((7-amino-2-( furan -2 -yl)- [1,2,4]triazolo[1,5-a] [ 1,3,5 ]triazine- 5-yl)amino)methyl)phenol ( JD005 , 4-(((7-Amino-2-( furan -2 - yl )-[1,2,4]triazolo[1,5-a][1,3 ,5]triazin-5-yl)amino)methyl)phenol) manufacturing

2-(furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazine-7-amine (2- (Furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) (50 mg, 0.18 mmol) Diisopropylethylamine (DIPEA) (0.16 mL, 0.89 mmol) and 4-hydroxybenzylamine (26 mg, 0.21 mmol) were added to a suspension containing methanol (1 mL). The reaction mixture was stirred overnight at room temperature. Then, a white solid product was obtained (10 mg, yield 17%) purified by column chromatography (1H-NMR (400 MHz, DMSO-D6) δ 9.20 (s, 1H), 7.82 (t, 2H), 7.11 ( d, 2H), 7.01 (t, 1H), 6.66 (s, 2H), 6.63 (m, 1H), 4.34 (d, 2H) MS [M+H]+=324.1, purity (LC) 98.07%) .

Example 2: Culture of pulp-derived stem cells

Dental pulp-derived stem cells (hDPSCs) were purchased and used from Lonza (PT-5012, Switzerland), xeno-free human MSC medium (StemMACS™ MSC Expansion Media Kit XF) containing antibiotic-antimycotic (anti-anti, final concentration 1X) , MACS) was used to culture the cells in a 5% CO ₂ , 37 °C incubator condition. The cells were lysed on the plate at 4000 cells/cm ² standard, and the medium was replaced every 3 days. Cells between 3 and 6 passages were used for the experiment.

Example 3: Measurement of cell viability

The pulp-derived stem cells were inoculated into a 96 well plate at 1 × 10 ³ cells/well. In order to confirm cytotoxicity when measuring viability, the compound was treated at each concentration, and cell viability was measured after culturing for 3 days. To measure cell viability, CCK-8 assay kit (Dongin Biotech, Daejeon) was used, and 10 μL of CCK-8 reagent was added to 100 μL of medium, incubated at 37 ° C for 2 hours, and then a microplate reader (Tecan, Sunrise) was used to measure absorbance at 450 nm.

Example 4: Induction of odontoblast differentiation

After inoculation at 1 × 10 ⁴ cells/well in a 48 well plate (SPL, Daejeon), differentiation was induced when the cells grew to 100% in the well. The medium used for differentiation induction was antibiotic-antimycotic (anti-anti, final concentration 1X), 10% FBS (GIBCO, 16000), 0.1 mM dexamethasone, 10 mM β-glycerophosphate (β-glycerophosphate) and 50 α-MEM medium (GIBCO) containing mM ascorbic acid was used, and JD005 was treated at concentrations of 1, 10, and 100 μM, respectively, and the medium was changed once every 3 days. Differentiation was performed for 14 days, and differentiation capacity was measured by Alizarin Red S staining (2% Alizarin Red S at pH 4.3 (Sigma-Aldrich, St Louis, MO, USA)). For the stained well, the plate was scanned with a scanner, and cell pictures were taken using a microscope, and after drying the dye well, 10% (w/v) cetylpyridinium chloride was added for quantitative analysis of staining. 10 mM sodium phosphate (pH 7.0) solution containing (Sigma-Aldrich) was added to each well by 300 μL, the dye was extracted, and the absorbance was measured at 570 nm using a microplate reader. did.

Example 5: RT-PCR analysis

RT-PCR was performed to confirm the expression of the odontoblast differentiation marker gene. On the 7th day after differentiation induction, the cells were recovered, RNA was extracted using Trizol (FavorPrep™ Tri-RNA reagent, Dongin Bio), and cDNA (bioneer, Daejeon) was synthesized. Gene expression was performed through RT-PCR (ABI 7500), and the primer sequences used in the experiment are shown in Table 1 below.

gene Forward Primer (5’→ 3’) Reverse Primer (5’→ 3’) DSPP GAA GAT GCT GGC CTG GAT AA TCT TCT TTC CCA TGG TCC TG DMP1 ACC TTT GGA GAC GAA GAC AAT GGC TGT CTT CAC TGG ACT GTG TGG TG ALP AGC TGA ACA GGA ACA ACG TGA CTT CAT GGT GCC CGT GGT C RUNX2 GGT TAA TCT CCG CAG GTC ACT CAC TGT GCT GAA GAG GCT GTT OPN AGC AGA GCG ACA CCC TAG A GCT CAT TGC TCT CAT CAT TGG C b-actin GGC ACC CAG CAC AAT GAA G TGC GGT GGA CGA TGG AGG

Example 6: Western blot analysis

Western blot analysis was performed to confirm odontoblast differentiation marker gene expression and cell signaling pathway. Protein was recovered using RIPA buffer (Intron) containing 1X protease/phosphatase inhibitor and western blot analysis was performed at a concentration of 30 μg. Primary antibodies used are listed in Table 2.

protein catalog number DMP-1 SC-73633, Santa cruz biotechnology DSPP SC-73632, Santa cruz biotechnology β-actin SC-, Santa cruz biotechnology Phospho SAMD - 1/5/8 AB3848-I, merchmillipore Total SMAD-1/5 PA5-80036, Thermofisher scientific

The following experimental examples were shown using the average value and standard deviation obtained by performing three independent experiments, and statistical differences between groups showed significance at P<0.05 or less.

Experimental example 1: JD005 to odontoblasts cytotoxicity and odontoblast Effect on Differentiation

An experiment was conducted to confirm the effect of JD005 synthesized in Example 1 during the differentiation period of pulp-derived mesenchymal stem cells into odontoblasts. Prior to cell differentiation, CCK-8 assay was performed to confirm the toxicity of JD005 to cells, and the degree of differentiation of odontoblasts was tested using concentrations of 1, 10, and 100 μM.

As a result, in FIG. 2A, when the concentration of JD005 was 100 μM, cell viability decreased by about 10%, but this was a very low rate, indicating that cell differentiation could be induced using this concentration. FIG. 2B And in FIG. 2C, it can be seen that the differentiation of odontoblasts increases whenever the concentration of JD005 increases. As a result of these experiments, it can be seen that as the concentration of JD005 increases, the differentiation of odontoblasts is also promoted.

Experimental example 2: JD005's of dental stem cells during eruption odontoblast differentiation marker Expression and degree of phosphorylation of SAMD 1, SAMD 5 and SAMD 8

JD005 was treated during the odontoblast differentiation period, and when treated at concentrations of 1, 10, and 100 μM, the expression of odontoblast differentiation marker genes and proteins was confirmed in the cells.

As a result, in FIGS. 3A to 3E, it was confirmed through RT-PCR that the increase in the differentiation marker gene according to the increase in the concentration of JD005 was confirmed, and in FIG. 3F, the expression of the protein used as the differentiation marker also increased through western blot. Confirmed. It was found that JD005 increased gene and protein expression as well as increased mineralization in differentiation into odontoblasts.

In addition, in FIG. 3G, the phosphorylation levels of SAMD 1, SAMD 5, and SAMD 8 were measured by confirming the signaling pathway of odontoblasts.

As a result, it was confirmed that the phosphorylation of SAMD 1, SAMD 5, and SAMD 8 was increased by JD005 treatment, and it was found that the differentiation of odontoblasts was promoted through this.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims (11)

A compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.
[Formula 1]

A composition for differentiating odontoblasts from dental stem cells, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The method according to claim 2, wherein the dental stem cells are dental pulp stem cells (DPSC), deciduous stem cells (Stem cell from human exfoliated deciduous teeth, SHED), dental papillary stem cells (Stem cells from the apical part of odontoblast cells from dental stem cells, characterized in that at least one selected from the group consisting of the papilla, SCAP), stem cells from the dental follicle (DFPC) and periodontal ligament stem cells (PDLSC) (odontoblast) A composition for differentiation. The composition for differentiating odontoblasts from dental stem cells according to claim 2, wherein the composition phosphorylates any one or more of SMAD 1, SMAD 5 and SMAD 8. A method for differentiating dental stem cells into odontoblasts, comprising treating dental stem cells with a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof to induce differentiation into odontoblasts.
[Formula 1]

A pharmaceutical composition for tooth or bone regeneration containing a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A health functional food for tooth or bone regeneration containing a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A health functional food for preserving or improving natural teeth containing a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A quasi-drug for preserving or improving natural teeth containing a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The method according to claim 9, wherein the quasi-drug is at least one formulation selected from the group consisting of toothpaste, mouthwash, mouthwash, gum, candy, oral spray, oral ointment, oral varnish, mouthwash, and gum massage cream. Quasi-drugs for preserving or improving natural teeth A first step in which 2-Furoic hydrazide and S-methylisothiouronium sulfate are mixed and stirred in a basic aqueous solution to obtain a precipitated material;
Water was added to the N“-(Furan-2-ylcarbonyl)carbonohydrazonic diamide obtained in the first step and stirred under microwave conditions. Then, a second step of obtaining a material by extracting and drying the reaction mixture in an organic solvent and then concentrating under reduced pressure;
5-(Furan-2-yl)-1H-1,2,4-triazol-3-amine (5-(Furan-2-yl)-1H-1,2,4-triazol obtained in the second step -3-amine) in a mixture of S,S'-dimethyl N-cyanodithioiminocarbonate (S,S'-dimethyl N-cyanodithioiminocarbonate), stirred in an inert gas atmosphere, and then put into an organic solvent to extract the substance. 3rd step to obtain;
2-(furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazine-7 obtained in the third step m- to -amine (2-(Furan-2-yl)-5-(methylthio)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) A fourth step of obtaining a substance by mixing with m-chloroperoxybenzoic acid under organic solvent conditions and then stirring; and
2-(furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazine- 7-amine (2-(Furan-2-yl)-5-(methylsulfonyl)-[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-amine) Sopropylethylamine (DIPEA) and 4-hydroxybenzylamine (4-hydroxybenzylamine) are mixed, and then stirred to obtain a material; a fifth step; a compound represented by Formula 1 comprising a method for producing a compound.
[Formula 1]

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