KR102157466B1 - Composition for preventing, improving or treating of bladder cancer comprising hexahydrotriazine derivatives - Google Patents

Abstract

The present invention relates to a hexahydrotriazine derivative, a preparation method thereof, and a composition for preventing, alleviating or treating bladder cancer, comprising the same. Since a compound represented by chemical formula 1 according to the present invention has excellent anti-bladder cancer activity, it may be useful for preventing, alleviating and treating bladder cancer.

Description

Composition for preventing, improving or treating of bladder cancer comprising hexahydrotriazine derivatives}

The present invention relates to a composition for preventing, improving or treating bladder cancer comprising a hexahydrotriazine derivative.

Cancer cells differ from normal cells in several ways. Normal cells proliferate according to their needs and where they are needed. Normal cells are attached to each other and appear to be together. In addition, normal cells destroy themselves if they are too old or damaged. However, cancer cells lack all of these functions and ultimately continue to divide, forming a tumor, a mass of cancer cells.

Bladder cancer is a malignant tumor of the bladder. The majority of bladder cancers are epithelial tumors derived from epithelial cells. Malignant epithelial tumors include transitional epithelial cell carcinoma, squamous cell carcinoma, and adenocarcinoma. In addition, it is divided into sarcoma derived from the muscles of the bladder, small cell carcinoma derived from nerve cells, malignant lymphoma, and metastatic cancer of the bladder in which cancer of other organs has metastasized to the bladder. According to data from the Central Cancer Registry released in 2019, 232,255 new cancers occurred in Korea in 2017, of which bladder cancer (C67) accounted for 1.9% of the total cancer incidence with 4,379 cases in both men and women.

In the case of bladder cancer, surgery or drug therapy is used depending on the symptoms.Surgery can cause complications of the digestive or urinary system, and the injection method of drugs into the bladder varies depending on the drug used, but chemotherapy drugs are absorbed by the bladder mucosa. It can cause systemic side effects and bladder irritation symptoms. It also affects normal cells to some extent, resulting in systemic complications. Overall, existing anti-bladder cancer drugs are accompanied by loss of appetite, nausea, vomiting, diarrhea, general weakness, and hair loss, and are highly cytotoxic and can destroy healthy cells. Accordingly, there is a need to develop anti-bladder cancer drugs that can minimize the side effects of existing anti-bladder cancer drugs and exhibit excellent anti-bladder cancer activity in trace amounts.

Accordingly, the present inventors devised a hexahydrotriazine derivative compound to synthesize a total of three derivatives, and confirmed that the hexahydrotriazine derivative compound exhibited excellent bladder cancer cell line killing effect and completed the present invention.

Korean Patent Registration No. 10-1797813

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

Another object of the present invention is to provide a method for preparing a compound represented by Chemical Formula 1.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of bladder cancer comprising a compound represented by Chemical Formula 1, or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a health functional food composition for preventing or improving bladder cancer, including the compound represented by Formula 1, or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a health food composition for preventing or improving bladder cancer comprising the compound represented by Formula 1, or a pharmaceutically acceptable salt thereof.

To achieve the above object,

The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ¹ is C _5-20 straight or branched chain alkyl;

R ² is a hydroxyl group or a thiol group).

In addition, the present invention, as shown in the following scheme 1,

Dissolving the compound represented by Formula 2 in a solvent, adding formaldehyde and a catalyst, and refluxing to synthesize a compound represented by Formula 3 (Step 1); And

Dissolving the compound represented by Formula 3 in step 1 in a solvent, adding the compound represented by Formula 4, and refluxing to synthesize a compound represented by Formula 1 (step 2);

It provides a method for preparing a compound represented by Chemical Formula 1 of claim 1 or a pharmaceutically acceptable salt thereof.

[Scheme 1]

(In the above Scheme 1,

R ¹ is C _5-20 straight or branched chain alkyl;

R ² is a hydroxyl group or a thiol group).

Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of bladder cancer comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

In addition, the present invention provides a health functional food composition for preventing or improving bladder cancer comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

Furthermore, the present invention provides a health food composition for preventing or improving bladder cancer comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

Since the compound represented by Formula 1 according to the present invention has excellent anti-bladder cancer activity, it may be useful for preventing, improving and treating bladder cancer.

1 is a graph showing the survival rate (%) of cells through the MTT assay of a bladder cancer cell line according to the concentration (0 to 2mM) treated with a hexahydrotriazine derivative according to the present invention.
2 is a graph showing the survival rate (%) of cells through the MTT assay of the bladder cancer cell line according to the concentration (0.5 or 1 mM) of the hexahydrotriazine derivative according to the present invention.

Hereinafter, the present invention will be described in detail.

Compound or a pharmaceutically acceptable salt thereof

The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ¹ is C _5-20 straight or branched chain alkyl;

R ² is a hydroxyl group or a thiol group.

Preferably,

R ¹ is C _5-15 straight or branched chain alkyl;

R ² may be a hydroxyl group or a thiol group.

In one embodiment of the present invention, preferred examples of the compound represented by Chemical Formula 1 according to the present invention include the following compound groups.

1) Hexahydrotriazinyl-trivalproyl-tris(3-mercaptopropanoic) anhydride;

2) hexahydrotriazinyl-trilauryl-tris(3-hydroxypropanoic) anhydride; or

3) Hexahydrotriazinyl-trivalproyl-tris(3-hydroxypropanoic) anhydride.

The chemical structural formulas of compounds that may be preferred examples of the compound represented by Formula 1 according to the present invention are summarized in Table 1 below.

Example Chemical structural formula One

2

3

The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The expression pharmaceutically acceptable salt is a concentration that is relatively non-toxic and harmless to the patient, and side effects caused by this salt do not degrade the beneficial efficacy of the basic compound of formula (1). It means inorganic addition salt. As these salts, inorganic acids and organic acids can be used as free acids, hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc. can be used as inorganic acids, and citric acid, acetic acid, lactic acid, maleic acid, fumarin Acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galucturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesol Ponic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid, malonic acid, and the like can be used. In addition, these salts include alkali metal salts (sodium salt, potassium salt, etc.) and alkaline earth metal salt (calcium salt, magnesium salt, etc.), and the like. For example, as an acid addition salt, acetate, aspartate, benzate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Gluceptate, Gluconate, Glucuronate, Hexafluorophosphate, Hybenzate, Hydrochloride/Chloride, Hydrobromide/Bromide, Hydroiodide/Iodide, Isethionate, Lactate, Malate, Male Eate, malonate, mesylate, methyl sulfate, naphthylate, 2-naphsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccha Rate, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, Potassium, sodium, tromethamine, zinc salts, and the like may be included, among which hydrochloride or trifluoroacetate is preferred.

The acid addition salt according to the present invention is a precipitate produced by dissolving the compound represented by Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid in a conventional method, for example, May be prepared by filtration and drying, or may be prepared by distilling a solvent and an excess of acid under reduced pressure and then drying or crystallizing in an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be made using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. In addition, the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

Furthermore, the present invention includes not only the compound of Formula 1 and a pharmaceutically acceptable salt thereof, but also possible solvates, hydrates, isomers, optical isomers, and the like that may be prepared therefrom.

Method for preparing a compound or a pharmaceutically acceptable salt thereof

The present invention is shown in the following scheme 1,

Dissolving the compound represented by Formula 2 in a solvent, adding formaldehyde and a catalyst, and refluxing to synthesize a compound represented by Formula 3 (Step 1); And

Dissolving the compound represented by Formula 3 in step 1 in a solvent, adding the compound represented by Formula 4, and refluxing to synthesize a compound represented by Formula 1 (step 2);

It provides a method for preparing a compound represented by Chemical Formula 1 of claim 1 or a pharmaceutically acceptable salt thereof.

[Scheme 1]

In Scheme 1 above,

R ¹ and R ² are as defined in Chemical Formula 1.

In the production method of the present invention, examples of the organic solvent of Step 1 or Step 2 include ethanol, tetrahydrofuran (THF), benzene, KOH/MeOH, MeOH, toluene, CH ₂ Cl ₂ , hexane, dimethylformamide ( DMF), diisopropyl ether, diethyl ether, dioxane, dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), acetone or chlorobenzene may be used, preferably ethanol as a solvent in step 1, Tetrahydrofuran (THF) may be used as a solvent.

Pharmaceutical composition for prevention or treatment of bladder cancer

The present invention provides a pharmaceutical composition for preventing or treating bladder cancer comprising a compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ¹ and R ² are as defined in claim 1.

In the pharmaceutical composition of the present invention, preferred examples of the compound represented by Formula 1 according to the present invention include the following compound groups.

1) Hexahydrotriazinyl-trivalproyl-tris(3-mercaptopropanoic) anhydride;

2) hexahydrotriazinyl-trilauryl-tris(3-hydroxypropanoic) anhydride; or

3) Hexahydrotriazinyl-trivalproyl-tris(3-hydroxypropanoic) anhydride.

In the pharmaceutical composition according to the present invention, the hexahydrotriazine derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered in various oral and parenteral dosage forms at the time of clinical administration. It can be prepared by using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used.

Formulations for oral administration include, for example, tablets, pills, hard/soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and troches.These formulations include diluents (e.g., lactose , Dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salt and/or polyethylene glycol). Tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and in some cases, boron such as starch, agar, alginic acid or sodium salt thereof. It may contain release or boiling mixtures and/or absorbents, colorants, flavoring agents, and sweetening agents.

The pharmaceutical composition containing the hexahydrotriazine derivative represented by Formula 1 as an active ingredient can be administered parenterally, and parenteral administration is by a method of injecting subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. All.

At this time, in order to formulate a formulation for parenteral administration, the hexahydrotriazine derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed in water with a stabilizer or buffer to prepare a solution or suspension, and this is an ampoule or It can be prepared in vial unit dosage form. The composition may be sterilized and/or contain adjuvants such as preservatives, stabilizers, hydrating agents or emulsification accelerators, salts and/or buffers for controlling osmotic pressure, and other therapeutically useful substances, which are conventional methods of mixing, granulation. It can be formulated according to the method of painting or coating.

In addition, the dose to the human body of the hexahydrotriazine derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition, and degree of disease. And, based on an adult patient weighing 70 Kg, it is generally 0.1-1000 mg/day, preferably 1-500 mg/day, and 1 day at regular intervals according to the judgment of a doctor or pharmacist. It may be administered in divided doses from once to several times.

Furthermore, the pharmaceutical composition of the present invention may be used alone for the prevention or treatment of bladder cancer, or in combination with surgery, hormone therapy, chemotherapy, and methods of using a biological response modifier.

Health functional food or health food composition for preventing or improving bladder cancer

The present invention provides a health functional food or health food composition for the prevention or improvement of bladder cancer comprising a compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ¹ and R ² are as defined in claim 1.

In the health functional food composition or health food composition according to the present invention, preferred examples of the compound represented by Formula 1 according to the present invention include the following compound groups.

1) Hexahydrotriazinyl-trivalproyl-tris(3-mercaptopropanoic) anhydride;

2) hexahydrotriazinyl-trilauryl-tris(3-hydroxypropanoic) anhydride; or

3) Hexahydrotriazinyl-trivalproyl-tris(3-hydroxypropanoic) anhydride.

The health functional food composition or health food composition according to the present invention is a compound represented by Formula 1, a food pharmaceutically acceptable salt thereof, or an optical isomer thereof for the purpose of preventing or improving bladder cancer. It can be added to functional foods or health foods.

There is no particular limitation on the type of food. Examples of health functional foods to which the compound according to the present invention can be added may be in the form of tablets, capsules, pills, or liquids, and examples of health foods include various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream , Dairy products, soups, ionic beverages, beverages, alcoholic beverages, gums, teas, and vitamin complexes, and include all health functional foods and health food compositions in the usual sense.

The health food and health functional food composition containing the compound according to the present invention may be added to food as it is or may be used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the compound according to the present invention may be appropriately determined depending on the purpose of use (for prevention or improvement). In general, the amount of the compound in health foods and health functional foods may be added in an amount of 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of maintaining health or for the purpose of health control, the amount may be less than the above range, and there is no problem in terms of safety, so the active ingredient may be used in an amount above the above range.

The health food and health functional food composition of the present invention is an essential ingredient in the indicated ratio, and there is no particular limitation on other ingredients, except for containing the compound according to the present invention, and various flavoring agents or natural carbohydrates, etc. It can contain as. Examples of the above-described natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 g of the health functional food composition of the present invention.

In addition to the above, the health food and health functional food composition containing the compound according to the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.) ), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the health food and health functional food composition of the present invention may contain flesh for the manufacture of natural fruit juice and fruit juice beverage and vegetable beverage.

These components may be used independently or in combination. Although the proportion of these additives is not so important, it is generally selected from 0.1 to about 20 parts by weight per 100 parts by weight of the health food and health functional food composition containing the active substance of the present invention.

<Preparation Example 1> Synthesis of 2-(4-isobutylphenyl)propanoic 2-propylpentanoic anhydride (2-(4-isobutylphenyl)propanoic 2-propylpentanoic anhydride) (G1)

Valproic acid (1.58 mL, 0.01 mol) and ibuprofen (2.06 g, 0.01 mol) were added to THF in a 250 mL one-neck round bottom flask sufficiently dried at room temperature using a Deanstark apparatus. It was refluxed using 20 mL as a solvent. After stirring for 24 hours, the water removed by Deanstark was checked, and the degree and completion of the reaction were checked by TLC, and the obtained solid was subjected to flash column chromatography (EA: n-Hexane = 1: 9 v/v). A lemon-colored liquid, 2-(4-isobutylphenyl)propanoic 2-propylpentanoic anhydride (0.89 g, 26.8%) was obtained.

2-(4-isobutylphenyl)propanoic 2-propylpentanoic anhydride:

Isolated yield: 45.1%; mp: sticky oil; R _f : 0.5 (TLC eluent EA: n-Hexane = 1: 9 v/v); MASS (70 eV), m/z (rel. intensity %); 332.2(100) 333.2(22.7) 334.2(2.5); ¹ H NMR (MeOH, 400 MHz): δ 0.96 (t, 9H), 1.38 (dd, 6H), 1.78 (m, 2H), 1.89 (m, 2H), 2.47 (s, 3H), 3.77 (t, 1H), 7.11 (t, 2H), 7.13 (t, 2H); ¹³ C NMR (MeOH, 100 MHz):δ 176.254(1C), 174.466(1C), 140.168(1C), 138.032(1C), 129.045(2C), 126.946(2C), 63.750(1C), 44.783(1C) , 34.598 (1C), 30.098 (2C), 24.868 (1C), 21.713 (2C), 20.440 (2C), 17.866 (2C), 13.347 (1C); Anal. Calcd. for C ₂₁ H ₃₂ O ₃ : C, 75.86; H, 9.70; O, 14.44; Found: C, 75.84; H, 9.72; O, 14.42

<Preparation Example 2> Synthesis of 2,2',2''-(1,3,5-triazinane-1,3,5-triyl) triacetic acid)

Add L-glycine (0.26 g, 3 mmol) to a 250 mL round-bottom flask that has been well-dried at room temperature, and dissolve in 25 mL ethanol, and then 5Å 20 mL of adsorbent, formaldehyde (0.9 mL, 3 mmol) ) And TEA (0.042 mL, 0.3 mmol) as a catalyst were added, and refluxed for 24 hours. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, the adsorbent was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; Methanol: ethyl acetate = 1: 3, v/v) was performed to obtain a yellow liquid 2,2',2''-(1,3,5-triazinane-1,3,5-triyl)triacetic acid. acid (0.02 g, yield; 7.0%) was obtained.

2,2',2''-(1,3,5-triazinane-1,3,5-triyl)triacetic acid):

Isolated yield: 7.0%; R _f : 0.50 (TLC eluent; Methanol: ethyl acetate = 1: 1, v/v); ¹ H NMR (D ₂ O, 400 MHz): δ 3.92 (s, 6H), 4.51 (s, 6H); ¹³ C NMR (D ₂ O, 100 MHz): δ 60.52, 77.88, 168.93; Anal. Calcd. for C ₉ H ₁₅ N ₃ O ₆ : C, 41.38; H, 5.79; N, 16.09; O, 36.75; Found: C, 41.38; H, 5.76; N, 16.07; O, 36.76

<Example 1> Hexahydrotriazinyl-trivalproyl-tris(3-mercaptopropanoic) anhydride; hexahydrotriazinyl-trivalproyl-tris(3-mercaptopropanoic) anhydride; Synthesis of compound E1)

<1-1> 2,2',2"-(1,3,5-triajinan-1,3,5-tris)tris(3-mercappropanoic acid) (2,2',2 Synthesis of "-(1,3,5-triazinane-1,3,5-triyl)tris(3-mercaptopropanoic acid))

L-cysteine (0.36 g, 3 mmol) was added to a well-dried 250 mL one necked round-bottom flask at room temperature, dissolved in 25 mL ethanol, and 5Å adsorbent (molecular sieves). ) 20 mL, formaldehyde (0.9 mL, 3 mmol) and TEA as a catalyst (0.042 mL, 0.3 mmol) were added, and refluxed for 24 hours. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, and the adsorbent (molecular sieves) was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; Methanol: ethyl acetate = 1: 3, v/v), a yellow liquid 2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris (3-mercaptopropanoic acid) (0.02 g, yield; 5.0%) was obtained.

2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris(3-mercaptopropanoic acid):

Isolated yield: 5.0%; R _f : 0.50 (TLC eluent; Methanol: ethyl acetate = 1: 1, v/v); ¹ H NMR (D ₂ O, 400 MHz): δ 3.44 (m, 6H), 4.36 (m, 3H), 4.48 (m, 6H); ¹³ C NMR (D ₂ O, 100 MHz): δ 32.64, 72.06, 75.97, 183.94

Anal. For C ₁₂ H ₂₁ N ₃ O ₆ S ₃ : C, 36.08; H, 5.30; N, 10.52; O, 24.03; S, 24.08; Found: C, 36.08; H, 5.36; N, 10.57; O, 24.06; S, 24.09

<1-2> Synthesis of Compound E1

Compound 2,2',2''-(1,3,5-triazinane-1,3 of Example 1-1 above) in a 250 mL one necked round-bottom flask well-dried at room temperature. ,5-triyl)tris(3-mercaptopropanoic acid) (0.99 g, 2.5 mmol) was added and dissolved in 20 mL THF, followed by 2-propylpentanoic acid (1.1 mL, 7.5 mmol) and 5Å adsorbent. (molecular sieves) 15 mL is added and refluxed for 24 hours. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, the adsorbent was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; EA: MeOH = 7: 1, v/v) was performed to obtain E1 (0.07 g, yield; 3.6%).

Isolated yield: 5.0%; R _f : 0.50 (TLC eluent; Methanol: ethyl acetate = 1: 1, v/v); ¹ H NMR (D ₂ O, 400 MHz): δ 3.44 (m, 6H), 4.36 (m, 3H), 4.48 (m, 6H); ¹³ C NMR (D ₂ O, 100 MHz): δ 32.64, 72.06, 75.97, 183.94

Anal. For C ₁₂ H ₂₁ N ₃ O ₆ S ₃ : C, 36.08; H, 5.30; N, 10.52; O, 24.03; S, 24.08; Found: C, 36.08; H, 5.36; N, 10.57; O, 24.06; S, 24.09

<Example 2> Synthesis of Hexahydrotriazinyl-trilauryl-tris(3-hydroxypropanoic) anhydride; Compound E2)

<2-1> 2,2',2"-(1,3,5-triajinan-1,3,5-triyl) tris(3-hydroxypropanoic acid) (2,2',2 Synthesis of''-(1,3,5-triazinane-1,3,5-triyl) tris(3-hydroxypropanoic acid)

Add L-serine (0.31 g, 3 mmol) to a 250 mL round-bottom flask that has been well-dried at room temperature, and dissolve in 25 mL ethanol, and then 5Å 20 mL of adsorbent, formaldehyde (0.9 mL, 3 mmol) And TEA (0.042 mL, 0.3 mmol) as a catalyst were added, and refluxed for 24 hours. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, the adsorbent was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; Methanol: ethyl acetate = 1: 3, v/v), a yellow liquid 2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris (3-hydroxypropanoic acid) (0.02 g, yield; 5.6%) was obtained.

2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris(3-hydroxypropanoic acid):

Isolated yield: 5.6%; R _f : 0.50 (TLC eluent; Methanol: ethyl acetate = 1: 1, v/v); ¹ H NMR (D ₂ O, 400 MHz): δ 3.83 (m, 3H), 4.07 (m, 6H), 4.33 (m, 6H); ¹³ C NMR (D ₂ O, 100 MHz):? 53.69, 68.98, 70.95, 178.77; Anal. Calcd. for C ₁₂ H ₂₁ N ₃ O ₉ : C, 14.03; H, 6.03; N, 11.96; O, 40.99; Found: C, 14.02; H, 6.06; N, 11.97; O, 40.96

<2-2> Synthesis of Compound E2

In a 250 mL one-neck round bottom flask well-dried at room temperature, 2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris(3- Add hydroxypropanoic acid) (0.99 g, 2.5 mmol), dissolve in 20 mL THF, add lauric acid (1.5 mL, 7.5 mmol) and 15 mL of 5Å adsorbent, and reflux for 24 hours. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, the adsorbent was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; EA: MeOH = 7: 1, v/v) was performed to obtain E2 (0.05 g, yield; 2.23%).

Isolated yield: 2.23%; mp: 595-596°C; R _f : 0.50 (TLC eluent; MeOH: ethylacetate = 1: 1, v/v); ¹ H NMR (CDCl ₃ ,400 MHz): δ 0.87 (t, 9H), 1.25 (s, 48H), 1.62 (s, 6H), 2.33 (t, 6H), 4.10 (m, 9H), 4.47 (m , 6H); ¹³ C NMR (CDCl ₃ , 400 MHz): δ 43.140, 52.379, 54.720, 58.893, 59.090, 59.121, 59.272, 59.386, 61.711, 63.591, 204.106, 206.219; Anal. Calcd. for C ₂₄ H ₃₉ N ₃ O ₁₂ : C, 51.33; H, 7.00; N, 7.48; O, 34.19; Found: C, 51.32; H, 7.46; N, 7.47; O, 34.16

<Example 3> Synthesis of Hexahydrotriazinyl-trivalproyl-tris (3-hydroxypropanoic) anhydride; Compound E3), hexahydrotriazinyl-trivalproyl-tris (3-hydroxypropanoic) anhydride

2,2',2''-(1,3,5-triazinane-1,3,5-triyl)tris(3-) of Example 2-1 above in a 250 mL one-necked round bottom flask dried well at room temperature. Add hydroxypropanoic acid) (0.87 g, 2.5 mmol), dissolve in 20 mL THF, add 2-propylpentanoic acid (1.1 mL, 7.5 mmol) and 15 mL of 5Å adsorbent, and reflux for 24 hours. do. After cooling at room temperature, the progress and completion of the reaction were checked by TLC, the adsorbent was removed using a glass filter, and the solvent was completely removed using a rotary vacuum concentrator. Flash column chromatography (eluent; EA: MeOH = 7: 1, v/v) was performed to obtain E3 (0.06 g, yield; 3.29%).

Isolated yield: 3.29%; R _f : 0.50 (TLC eluent; MeOH: ethylacetate = 1: 1, v/v); ¹ H NMR (D ₂ O, 400 MHz): δ 1.00-1.084 (m, 18H), 1.41-1.92 (m, 24H), 2.49 (m, 3H), 3.83 (m, 3H), 3.94 (m, 6H) ), 4.68 (m, 6H); ¹³ C NMR (D ₂ O, 400 MHz): δ 14.488, 21.170, 24.687, 35.117, 45.802, 69.818, 72.140, 169.183, 176.138; Anal. Calcd. for C ₃₆ H ₆₃ N ₃ O ₁₂ : C, 59.24; H, 8.70; N, 5.76; O, 26.30; Found: C, 59.22; H, 8.70; N, 5.77; O, 26.30

<Experimental Example 1> Evaluation of apoptosis effect on bladder cancer cell lines

<1-1> Cell culture

In order to evaluate the anti-bladder cancer effect of the hexahydrotriazine derivative compounds according to Examples 1 to 3, apoptosis effect analysis was performed on a bladder cancer cell line (5637) purchased from the American Type Culture Collection (ATCC).

The bladder cancer cell line (5637) was cultured in RPMI supplemented with 10% FBS and 1% penicillin/streptomycin (100 U/ml). The cells were maintained at 37° C. in a humidified atmosphere of 5% CO ₂ .

<1-2> Cell activity analysis through MTT assay

MTT analysis was performed to confirm the viability of 5637 cells for the hexahydrotriazine derivative compounds according to Examples 1 to 3. The bladder cancer cell line (5637) was seeded in a 96-well plate at 3 × 10 ³ cells/well. The next day, the compounds of Examples 1 to 3 and Preparation Example 1 dissolved in deionized water were treated at a concentration of 0 mM to 2 mM for 24 hours. After 24 hours, the medium was replaced with 10 mL MTT solution (5 mg MTT / 1 mL 1 × PBS). After incubation for 4 hours, 100 mL of DMSO was added to measure the absorbance at 550 nm. Cell viability (%) was calculated as (absorbance value of cells treated with chemical substances / absorbance value of untreated cells) × 100.

The results are shown in Figs. 1 and 2, and the x-axis of the graph shows the concentration, and the y-axis shows the cell survival rate when the synthesized substance is administered to cancer cells. When the cell survival rate of cancer cells is 50% or less, it can be determined that there is an anticancer effect, and as shown in FIG. 1, the cancer cell survival rate as the concentration of both of the compounds of Examples 1 to 3 and Preparation Example 1 (control group; compound G1) increases Since this decreased, it was confirmed that apoptosis of the bladder cancer cell line (5637) was induced in a concentration-dependent manner, and as shown in FIGS. 1 and 2, in the case of Example 1 (E1), high apoptosis was induced even at a low concentration, and 0.5 At a concentration of mM or more, it was confirmed that the cancer cell killing effect was close to 100%, indicating an excellent anti-bladder cancer effect.

In addition, Example 3 (E3) was found to have higher solubility in solvent than other substances, so that the experiment was possible at an accurate concentration, and the reproducibility of the result was high. In addition, although it had a weak apoptosis effect compared to Example 1 (E1), it showed meaningful results at a concentration of 0.5 mM or more, and showed a very high anticancer effect compared to Preparation Example (G1). While Example 1 (E1) and Example 3 (E3) showed strong anti-cancer effects as shown in FIGS. 1 and 2, the result value for Example 2 (E2) was constant in concentration as shown in FIG. The results were not shown, and were not separately shown in FIG. 1.

It was found that the hexahydrotriazine derivative compound according to Example 1 of the present invention has an anti-bladder cancer effect of specifically acting on bladder cancer cells to kill cells.

The above results suggest that the compound according to the embodiment of the present invention has anti-bladder cancer activity.

<Formulation Example 1> Preparation of pharmaceutical formulation

<1-1> Preparation of powder

2 g of compound of formula 1

1 g lactose

After mixing the above ingredients, the powder was prepared by filling the airtight cloth.

<1-2> Preparation of tablets

100 mg of the compound of formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet preparation method.

<1-3> Preparation of capsules

100 mg of the compound of formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, the gelatin capsules were filled according to a conventional capsule preparation method to prepare a capsule formulation.

<1-4> Preparation of injection solution

Compound of Formula 1 10 ㎍/㎖

Until diluted hydrochloric acid BP pH 3.5

Sodium chloride BP for injection up to 1 ml

Dissolve the compound of formula 1 according to the present invention in an appropriate volume of sodium chloride BP for injection, adjust the pH of the resulting solution to pH 3.5 with dilute hydrochloric acid BP, adjust the volume with sodium chloride BP for injection, and sufficiently Mixed. The solution was filled in a 5 ml type I ampoule made of transparent glass, the glass was dissolved and sealed under an upper grid of air, and sterilized by autoclaving at 120° C. for 15 minutes or longer to prepare an injection solution.

<Formulation Example 2> Preparation of health functional food and health food

<2-1> Manufacture of health functional food

100 mg of the compound of formula 1

Vitamin mixture right amount

Vitamin A acetate 70 μg

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

1.7 mg of nicotinic acid amide

50 μg folic acid

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is a mixture of ingredients suitable for a health functional food in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules are prepared, and can be used for preparing a health functional food composition according to a conventional method.

<2-2> Production of health functional beverages

100 mg of the compound of formula 1

100 mg citric acid

100 mg oligosaccharide

Plum Concentrate 2 mg

100 mg taurine

Total 500 mL with purified water

에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 1 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. 상기 조성비는 비교적 기호 음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.After mixing the above ingredients according to a conventional health drink manufacturing method, 85

After stirring and heating at, the resulting solution is filtered and obtained in one sterilized container, sealed and sterilized, and then stored in a refrigerator, and then used to prepare the health drink composition of the present invention. The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the intended use.

<2-3> Manufacture of dairy products

0.01-1 parts by weight of the hexahydrotriazine derivative compound of Formula 1 of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-4> Manufacturing of Seonsik

Brown rice, barley, glutinous rice, and adlay were gelatinized by a known method, dried, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder. The dry powder of the grains and seeds prepared above and the hexahydrotriazine derivative compound of Formula 1 of the present invention were mixed in the following ratio to prepare.

Cereals (34 parts by weight of brown rice, 19 parts by weight of barley, 20 parts by weight of barley),

Seeds (perilla 7 parts by weight, black beans 8 parts by weight, black sesame 7 parts by weight),

A compound of formula 1 (2 parts by weight),

Ganoderma lucidum (1.5 parts by weight), and

Rehmannia (1.5 parts by weight).

Claims (10)

A pharmaceutical composition for preventing or treating bladder cancer comprising a compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1,
R ¹ is C _5-20 straight or branched chain alkyl;
R ² is a hydroxyl group or a thiol group). delete delete delete delete delete A health functional food composition for preventing or improving bladder cancer comprising the compound represented by Formula 1 of claim 1 or a pharmaceutically acceptable salt thereof. The method of claim 7,
The health functional food is a health functional food composition for preventing or improving bladder cancer, characterized in that the food in the form of a tablet, capsule, pill or liquid. A health food composition for preventing or improving bladder cancer comprising a compound represented by Formula 1 of claim 1 or a pharmaceutically acceptable salt thereof. The method of claim 9,
The health food is selected from various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ion drinks, beverages, alcoholic beverages, gum, tea, and vitamin complexes. Health food composition for improvement.

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