KR20210106156A - Composition for preventing or treating of leukemia comprising a benzofuran-pyrazole derivative compound that inhibits ASH1Lhistone methyltransferase activity - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating leukemia comprising a benzofuran-pyrazole derivative, wherein the composition according to the present invention effectively inhibits ASH1L histone methylase activity, and has an effect of promoting the death of leukemia cells since the ASH1L histone methylase activity is reduced, thereby being able to be usefully used for preventing or treating leukemia.

Description

BACKGROUND ART A composition for preventing or treating leukemia comprising a benzofuran-pyrazole derivative compound that inhibits ASH1L histone methylation enzyme activity.

The present invention relates to a pharmaceutical composition for preventing or treating leukemia comprising a benzofuran-pyrazole derivative, and more particularly, to a compound that inhibits ASH1L histone methylation enzyme activity.

Acute leukemia is a disease in which abnormal leukocyte progenitor cells or platelet progenitor cells are excessively produced due to a bone marrow abnormality. This is called acute myeloid leukemia (AML). Since abnormal white blood cells increase and occupy the place of hematopoiesis, normal white blood cells, red blood cells, platelets, etc. cannot be formed, so infection or bleeding occurs easily, and if not treated, death occurs within several months. Although the survival rate of acute leukemia in infants has improved significantly with recent advances in chemotherapy, the survival rate in adults is still low.

Acute myeloid leukemia is a malignant tumor arising from stem cells of non-lymphocytes or myeloid leukocytes produced in the bone marrow. Genetic mutations in hematopoietic stem cells cause myeloid progenitors to stop differentiation at various stages, and immature myeloblasts become single cells. It is a hematopoietic tumor that proliferates in a group (monoclonal). Symptoms of bone marrow dysfunction, such as anemia, fever, increased infectivity, and bleeding tendency, are also present, and there are cases of long-term infiltration of tumor cells, such as splenomegaly and lymph node swelling.

As a standard method for treating leukemia, chemotherapy, hematopoietic stem cell transplantation, radiation therapy, etc. are included, and in the case of chemotherapy, a method of using two or more anticancer agents in combination is usually included. The ideal chemotherapy would be that the anti-leukemia drug should show a selective effect only on leukemia cells without inhibiting normal hematopoiesis and without causing other harmful side effects. However, although most anti-leukemia drugs can kill leukemia cells close to the ideal state, they also inhibit normal hematopoiesis and cause other harmful side effects, so there is a limit to the treatment of leukemia. In addition, antitumor effects are weak in drug-resistant leukemia cells, side effects may occur, and sufficient chemotherapy may not be administered in some cases.

ASH1L is a histone lysine methyl transferase (KMTase) specific for histone H3, lysine 36, which plays a variety of roles in several types of cancer. During development, ASH1L activates the HOX gene and regulates the self-renewal capacity of long-term hematopoietic stem cells (LT-HSCs). The HOX gene is a carcinogen in various cancer types. For example, overexpression of HOXA9 is associated with a poor prognosis in acute myeloid leukemia. In addition, ASH1L is overexpressed in various cancers, including breast and thyroid cancer (Rogawski, David, et al. "The role of the ASH1L SET domain in leukemogenesis." (2017): 1373-1373.). In a recent study, it was reported that ASH1L induces leukemia in MLL by recruiting a mixed-lineage leukemia (MLL) epigenetic regulator (Zhu, Li, et al. "ASH1L links histone H3 lysine 36 dimethylation to MLL leukemia." 　 Cancer discovery 6.7 (2016): 770-783.). In addition, ASH1L, which regulates leukemia cell growth in CALM-AF10 and MLL-AF9-driven leukemia, plays an important role in leukemia expressing high HOX (Rogawski, David. The function of the ASH1L histone methyltransferase in cancer: A chemical biology approach. Diss. 2018.)

On the other hand, the high-speed ligand screening method using DSF (differential scanning fluorimetry) has the advantage of being able to easily perform large-scale, high-speed screening from natural mixtures to single compounds by measuring the change in the protein denaturation transition temperature using a compound that strongly binds to a target protein. have. Since DSF screening can directly search for compounds that bind to a target protein at the molecular level, it has the advantage of being developed as an innovative or natural drug, that is, leading materials can be secured in a short time.

Accordingly, the present inventors have completed the present invention by identifying a compound capable of preventing or treating acute myeloid leukemia by inhibiting the activity of ASH1L.

Korean Patent Publication No. 10-2019-0049584

1. Zhu, Li, et al. "ASH1L links histone H3 lysine 36 dimethylation to MLL leukemia." Cancer discovery　6.7 (2016): 770-783.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating leukemia.

Another object of the present invention is to provide a health functional food composition for preventing or improving leukemia.

Another object of the present invention is to provide a health food composition for preventing or improving leukemia.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for preventing or treating leukemia comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

(in Formula 1 above

R ¹ and R ² are independently hydrogen, halogen, hydroxy or halogenated C _1-5 straight-chain or branched alkyl).

In addition, the present invention provides a health functional food composition for preventing or improving leukemia 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 Formula 1 of claim 1).

Furthermore, the present invention provides a health food composition for preventing or improving leukemia 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 Formula 1 of claim 1).

The present invention is a novel composition having an effect of inhibiting ASH1L histone methylase activity, wherein the composition according to the present invention effectively inhibits ASH1L histone methylase activity, and the ASH1L histone methylase activity is inhibited to promote death of leukemia cells Therefore, it can be usefully used to prevent or treat leukemia.

1 is a result of confirming the melting point (Tm) of the compounds of Comparative Examples 1 to 7 for ASH1L-MRG15 using differential scanning fluorometry (DSF).
2 is a result of confirming the melting point (Tm) of the compounds of Comparative Examples 8 to 10 and Example 1 for ASH1L-MRG15 using differential scanning fluorescence spectroscopy (DSF).
3 is a result of confirming the melting point (Tm) of the compounds of Comparative Examples 11 to 14 for ASH1L-MRG15 using differential scanning fluorescence spectroscopy (DSF).
4 is a result of confirming the melting point (Tm) of the compounds of Comparative Examples 15 to 18 for ASH1L-MRG15 using differential scanning fluorescence spectroscopy (DSF).
5 is a result of measuring the ASH1L histone methylation enzyme activity of Comparative Examples 1 to 18 and Example 1 (first lane: enzyme ASH1L-MRG15, second lane: substrate nucleosome, compound concentrations of 10 μM and 100 μM, respectively) activation).

Hereinafter, the present invention will be described in detail.

Pharmaceutical composition for preventing or treating leukemia

The present invention provides a pharmaceutical composition for preventing or treating leukemia comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

(in Formula 1 above

R ¹ and R ² are independently hydrogen, halogen, hydroxy or halogenated C _1-5 straight-chain or branched alkyl).

Preferably, R ¹ and R ² may be independently hydrogen, halogen, hydroxy or halogenated C _1-3 linear or branched alkyl, more preferably independently hydrogen, halogen, hydroxy or trihalo may be methyl.

In the pharmaceutical composition of the present invention, the compound represented by Formula 1 is

3-(2,3-dihydrobenzofuran-5-yl)-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol; can be

In the pharmaceutical composition of the present invention, the leukemia is acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, or chronic lymphocytic leukemia. It may be, preferably, acute myeloid leukemia (Acute myeloid leukemia).

In the pharmaceutical composition of the present invention, the composition has the effect of promoting the death of leukemia cells by inhibiting the activity of ASH1L histone methylation enzyme.

The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The expression pharmaceutically acceptable salt refers to any organic or organic compound of the basic compound of formula (1), in which the concentration has an effective action that is relatively non-toxic and harmless to the patient, and the side effects due to the salt do not reduce the beneficial efficacy of the basic compound of formula (1). means inorganic addition salts. For these salts, inorganic acids and organic acids can be used as free acids, and hydrochloric acid, hydrobromic 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, etc. can be used as organic acids. Acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid Phonic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid may be used. Further, 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, acid addition salts include acetate, aspartate, benzate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Gluceptate, gluconate, glucuronate, hexafluorophosphate, hebenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, malate ate, malonate, mesylate, methylsulfate, naphthylate, 2-naphsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate Late, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, Potassium, sodium, tromethamine, zinc salt and the like may be included, of which hydrochloride or trifluoroacetate is preferable.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound represented by Formula 1 in an organic solvent, for example, methanol, ethanol, acetone, methylene chloride, acetonitrile, or the like, and adding an organic or inorganic acid to the precipitate. It can be prepared by filtration and drying, or by drying or crystallization in an organic solvent after distilling the solvent and excess acid under reduced pressure.

In addition, a pharmaceutically acceptable metal salt may be prepared 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. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding silver salt is also 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 or a pharmaceutically acceptable salt thereof, but also all possible solvates, hydrates, isomers, and optical isomers that can be prepared therefrom.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention may be administered in various oral and parenteral formulations during clinical administration, and when formulated, commonly used fillers, extenders, binders, and wetting agents. , is prepared using a diluent or excipient such as a disintegrant or surfactant.

Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, and these solid preparations include at least one compound represented by Formula 1 of the present invention or a pharmaceutically acceptable salt thereof. It is prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, or syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. can

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspension solutions, emulsions, lyophilized formulations, suppositories, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, etc. may be used.

In addition, the effective dose to the human body of the compound 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 status and disease degree, and generally as about 0.001-100 mg/kg/day, preferably 0.01-35 mg/kg/day. Based on an adult patient weighing 70 kg, it is generally 0.07-7000 mg/day, preferably 0.7-2500 mg/day, and once a day at regular time intervals according to the judgment of a doctor or pharmacist It may be administered in several divided doses.

Health functional food or health food composition for preventing or improving leukemia

The present invention provides a health functional food or health food composition for preventing or improving leukemia 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 Formula 1 of claim 1).

In the composition of the present invention, the leukemia may be acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, or chronic lymphocytic leukemia. , preferably acute myeloid leukemia.

In one embodiment of the present invention, the composition has the effect of promoting the death of leukemia cells by inhibiting the activity of ASH1L histone methylation enzyme.

The term "health functional food" as used in the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, pills or liquids using raw materials or ingredients having useful functions in the human body. Here, the term 'functionality' refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention is not particularly limited in the type of conventional food. Examples of foods to which the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention can be added include various drinks, meat, sausage, bread, candies, snacks, noodles, ice cream, dairy products, soups, and ionic beverages. , beverages, alcoholic beverages, gum, tea, vitamin complex, and the like, and includes both health food and health functional food in the ordinary sense.

The health food and health functional food composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof according to the present invention can be added to food as it is or used together with other food or food ingredients, and can be used in a conventional method. can be used appropriately. The mixing amount of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be suitably determined depending on the purpose of its use (for prevention or improvement). In general, the amount of the composition in health food and health functional food may be added in an amount of 0.1 to 90 parts by weight based on the total weight of the food. However, in the case of long-term ingestion for health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the compound represented by Formula 1 or its pharmaceutically acceptable The salt may be used in an amount above the above range.

The health food and health functional food composition of the present invention is not particularly limited in other ingredients except for containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in the present invention as an essential ingredient in the indicated ratio. It may contain various flavoring agents or natural carbohydrates as additional ingredients. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional 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 (taumatine, 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 health functional food composition of the present invention.

In addition to the above, the health food and health functional food composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and natural flavoring agents, etc. used in flavoring, coloring 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, carbonated beverages Carbonating agents and the like may be contained. In addition, the health food and health functional food composition of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage.

These components may be used independently or in combination. Although the ratio of these additives is not so important, it is selected from the range of 0.1 to about 20 parts by weight per 100 parts by weight of the health food and health functional food composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention. it is common to be

The present invention will be described in more detail through the following examples. However, the following examples are only for specifying the contents of the present invention, and the present invention is not limited thereto.

<Preparation example>

The compounds of Table 1 below were purchased from Fisher Science Korea as Maybridge products.

division compound name structure Comparative Example 1
(21B07) 4-(trifluoromethylthio)phenol

Comparative Example 2
(21C08) 2-(2,3-dichlorophenyl)thiazole-4-carboxylic acid

Comparative Example 3
(25G11) ethyl 2-(pyrrolidin-1-yl)acetate

Comparative Example 4
(25A11) (4-((1H-pyrazol-1-yl)methyl)phenyl)methanol

Comparative Example 5
(29E02) 6-(4-chloro-3-methylphenoxy)pyridin-3-amine

Comparative Example 6
(30B02) 2-phenyl-5-(trifluoromethyl)-1,2-dihydropyrazol-3-one

Comparative Example 7
(30H02) (5-tert-butylthiophen-2-yl)(pyrrolidin-1-yl)methanone

Comparative Example 8
(36H04) 2-ethyl-4-methyl-1H-imidazole

Comparative Example 9
(37E08) 4-phenyl-5-(trifluoromethyl)thiophene-2-carboxylic acid

Comparative Example 10
(37E09) 3-amino-4-phenyl-5-(trifluoromethyl)thiophene-2-carboxylic acid

Comparative Example 11
(38F07) 1-(2-(4-(trifluoromethyl)phenyl)thiazol-4-yl)ethanone

Comparative Example 12
(35D10) 3-phenyl-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol

Comparative Example 13
(35E11) 5-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)thiophene-2-carboxylic acid

Comparative Example 14
(36H02) (E)-2-(hydroxyimino)-N-(3-(trifluoromethyl)phenyl)propanamide

Comparative Example 15
(36E04) 2-hydroxy-6-(thiophen-2-yl)-4-(trifluoromethyl)nicotinonitrile

Comparative Example 16
(37C06) ethyl 2-(4-(trifluoromethyl)phenylamino)acetate

Comparative Example 17
(37G03) 7-hydroxy-4-(trifluoromethyl)-2H-chromen-2-one

Comparative Example 18
(37G08) 4-phenyl-3-(trifluoromethyl)-1H-pyrazole

Example 1
(38H04) 3-(2,3-dihydrobenzofuran-5-yl)-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol

<Experimental Example 1> Differential scanning fluorescence measurement

A thermal shift assay that measures the binding between a target protein and a compound using differential scanning fluorimetry (DSF) is used. That is, the measurement of the compound binding to the protein uses the fact that when the compound is bound, the melting point (Tm) is increased compared to the unbound protein. DSF was performed using an Applied Biosystems® StepOnePlus ^™ Real-Time PCR System (Thermo Fisher Scientific Co.). Samples were prepared in a final volume of 20 μl as follows: 1 mg ml ^-1 ASH1L-MRG15 (MORF-related gene 15 protein) complex (2 μl), 1 mM compound (2 μl, 2%) dissolved in 20% DMSO DMSO), 2.5Х SYPRO orange (2.5 μl), 10Х buffer (2 μl), distilled H ₂ O (11.5 μl). Fluorescence (λ _ex =580 nm, λ _em =623 nm) was measured at a rate of 0.05°C/s from 25°C to 99°C to monitor the protein unwinding state, and the melting point was obtained therefrom. As a result, the Tm of the ASH1L-MRG15 complex was 47.5°C.

Based on the annealing intermediate temperature (Tm) of the ASH1L-MRG15 complex obtained from differential scanning fluorometry, the change in Tm value was measured when the compound was added. As shown in FIGS. 1 to 4, Example 1 of the present invention It was confirmed that the Tm value significantly changed when the compound was added.

<Experimental Example 2> Measurement of the activity of histone methylation enzyme of ASH1L

Tritiated form of S-adenosyl methionine (SAM), a methyl donor cofactor of ASH1L, in order to measure the activity of histone methylation enzyme of ASH1L of the compounds of Comparative Examples 1 to 18 and Example 1 according to the experimental results of Experimental Example 1 The transfer of H ³ -SAM with methyl of ASH1L to histone H3, a substrate of ASH1L, is measured using the radio-autograph method. ASH1L enzyme and an oligo-nucleosomal array consisting of several nucleosomes (histone + DNA) as a substrate and H ³ -SAM were reacted, and the compounds of Comparative Examples 1 to 18 and Example 1 were added thereto, and the reaction was carried out at 37 ° C. Then, histones were separated through SDS-PAGE, the gel was transferred to a PVDF membrane, and the activity of histone methylation enzyme was measured by exposing it to a phosphor-image plate.

For each compound at final concentrations of the enzyme ASH1L-MRG15 400 nM, the substrate nucleosome 120 ng/μl and the compound Control (0 uM), 10 μM, 100 μM, the degree of inhibition of the enzyme activity was measured through autoradiograph. As a result, as shown in FIG. 5 , it was confirmed that the compound of Example 1 exhibited excellent inhibitory activity.

Preparation example of drug

The compound of Formula 1 according to the present invention can be formulated in various forms depending on the purpose. The following exemplifies several formulation methods containing the compound of Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Pharmaceutical Preparation Example 1> Preparation of powder

compound of formula 1 2 g

lactose 1 g

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

<Pharmaceutical Preparation Example 2> Preparation of tablets

compound of formula 1 100 mg

corn starch 100 mg

lactose 100 mg

magnesium stearate 2 mg

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

<Pharmaceutical Preparation Example 3> Preparation of capsules

compound of formula 1 100 mg

corn starch 100 mg

lactose 100 mg

magnesium stearate 2 mg

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

<Pharmaceutical Preparation Example 4> Preparation of injection

compound of formula 1 10 μg/ml

dilute hydrochloric acid BP until 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 clear glass, sealed under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120° C. for 15 minutes or more to prepare an injection solution.

<Pharmaceutical Preparation Example 5> Preparation of nasal absorbent (Nasal spray)

compound of formula 1 1.0 g

sodium acetate 0.3 g

methylparaben 0.1 g

Propylparaben 0.02 g

sodium chloride appropriate amount

HCl or NaOH pH adjustment appropriate amount

Purified water appropriate amount

According to a conventional method for preparing nasal absorbents, 3 mg of the compound of Formula 1 per 1 mL of saline (0.9% NaCl, w/v, solvent is purified water) is prepared, filled in an opaque spray container, and sterilized to An absorbent was prepared.

<Pharmaceutical Preparation Example 6> Preparation of liquid preparation

compound of formula 1 100 mg

Lee Seonghwadang 10 g

mannitol 5 g

Purified water appropriate amount

According to a conventional liquid preparation method, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, and purified water is added to adjust the total to 100 mL, then filled in a brown bottle and sterilized to prepare a liquid preparation did.

Manufacturing example of health food

The compound of Formula 1 according to the present invention can be prepared into various types of health food depending on the purpose. The following exemplifies a method for producing several health foods containing the compound of Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Health food production example 1> Production of dairy products

0.01-1 parts by weight of the 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.

<Health food production example 2> Preparation of wire

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared into powder having a particle size of 60 mesh with a grinder. The compound of Formula 1 of the present invention was concentrated under reduced pressure in a vacuum concentrator to obtain a dry powder. It was prepared by mixing the grains, seeds and dry powder of the compound of Formula 1 prepared above in the following ratios.

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

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

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

Reishi (1.5 parts by weight), and

Rehmannia (1.5 parts by weight).

Manufacturing example of health functional food

The compound of Formula 1 according to the present invention can be prepared into various types of health functional food depending on the purpose. The following exemplifies the manufacturing method of several health functional foods containing the compound of Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Health functional food manufacturing example 1> Manufacture of health functional food

compound of formula 1 100 mg

vitamin mixture appropriate amount

vitamin A acetate 70 μg

vitamin E 1.0 mg

vitamin B1 0.13 mg

vitamin B2 0.15 mg

vitamin B6 0.5 mg

vitamin B12 0.2 μg

vitamin C 10 mg

biotin 10 μg

Nicotinamide 1.7 mg

folic acid 50 μg

Calcium Pantothenate 0.5 mg

mineral mixture appropriate amount

ferrous sulfate 1.75 mg

zinc oxide 0.82 mg

magnesium carbonate 25.3 mg

monobasic potassium phosphate 15 mg

Dibasic Calcium Phosphate 55 mg

Potassium Citrate 90 mg

calcium carbonate 100 mg

magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food, but the composition is mixed 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 can be prepared and used in the preparation of a health functional food composition according to a conventional method.

<Health functional food manufacturing example 2> Manufacture of health functional beverage

compound of formula 1 100 mg

citric acid 100 mg

oligosaccharide 100 mg

Plum Concentrate 2 mg

Taurine 100 mg

Purified water is added to 500 mL

After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in one sterilized container, sealed and sterilized, and then refrigerated. used in the manufacture of health beverage compositions of Although the composition ratio is a composition that is relatively suitable for a beverage of preference in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, demand country, and use purpose.

So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in modified forms without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than in the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (13)

A pharmaceutical composition for preventing or treating leukemia comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1 above
R ¹ and R ² are independently hydrogen, halogen, hydroxy or halogenated C _1-5 straight-chain or branched alkyl). According to claim 1,
wherein R ¹ and R ² are independently hydrogen, halogen, hydroxy, or halogenated C _1-3 linear or branched alkyl; Wherein R ¹ And R ^{2 Are} independently hydrogen, halogen, hydroxy or trihalomethyl pharmaceutical composition, characterized in that. According to claim 1,
The compound represented by Formula 1 is,
3-(2,3-dihydrobenzofuran-5-yl)-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol; A pharmaceutical composition, characterized in that According to claim 1,
The leukemia is acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, or chronic lymphocytic leukemia. 6. The method of claim 5,
The leukemia is a pharmaceutical composition, characterized in that acute myeloid leukemia (Acute myeloid leukemia). According to claim 1,
The composition is a pharmaceutical composition, characterized in that inhibiting the activity of ASH1L histone methylation enzyme. A health functional food composition for preventing or improving leukemia 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 Formula 1 of claim 1). 9. The method of claim 8,
The health functional food is a health functional food composition for preventing or improving leukemia, characterized in that it is in the form of tablets, capsules, powders, granules, pills or liquids. A health food composition for preventing or improving leukemia 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 Formula 1 of claim 1). 11. The method of claim 10,
The health food is selected from various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea, and vitamin complexes. Prevention or improvement of leukemia, characterized in that health food composition for use. 11. The method of claim 8 or 10,
The leukemia is acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, or chronic lymphocytic leukemia. Composition, characterized in that it is. 13. The method of claim 12,
The leukemia is a composition, characterized in that acute myeloid leukemia (Acute myeloid leukemia).

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