KR101831981B1 - Composition for preventing, improving or treating cancer comprising Sageretia thea extract or its fraction as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating cancer comprising a Sageretia thea extract or a fraction thereof as an effective component, wherein the Sageretia thea extract and fractions for each solvent thereof reduce the cell survival rate of breast cancer cells, exhibit a cell suicide inducing effect, and can secure stability by being derived from plants, thereby being used for health functional foods and medicines for preventing, improving or treating the breast cancer.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing, ameliorating or treating cancer, which comprises as an active ingredient an extract of the same or a fraction thereof,

The present invention relates to a composition for preventing, ameliorating or treating a cancer containing an extract of the same or a fraction thereof as an active ingredient.

Cancer is one of the incurable diseases that humanity needs to solve. In the world, huge capital is invested in development to heal it globally. In Korea, it is the first disease among the causes of death, It is diagnosed and more than 60,000 people are dying. Smoking, ultraviolet rays, chemicals, food, and other environmental factors are among the carcinogens that cause cancer, but the development of therapeutic agents is difficult due to the various causes of the cancer, and the effect of the therapeutic agent varies depending on the site . Current therapeutic agents have considerable toxicity and can not selectively remove cancer cells. Therefore, there is a desperate need for the development of effective anti-cancer drugs that are less toxic.

Sageetia thea is an evergreen shrub native to southern coast of Korea and coastal area of Jeju island. It is mainly used for bonsai in Japan and USA. Leaves are small and have thorns and are sometimes called pedicel trees. It is said that it is efficacious to drink tea made with this leaf in cold, but there is little research on this plant.

Korean Patent Registration No. 0969134 discloses a method for separating a fraction containing a large amount of useful flavonoids and a novel flavonoid substance from the surface of a Sambrook tree. However, the present invention provides a method for preventing, Compositions for improvement or treatment have not yet been disclosed.

The present invention has been made in view of the above-described needs, and it is an object of the present invention to provide a method for treating breast cancer cells, which comprises treating the Sageretia thea extract and the solvent- (apoptosis), and thus the present invention has been completed.

In order to achieve the foregoing object, the present invention is homologous tree (Sageretia The present invention provides a pharmaceutical composition for preventing or treating cancer containing an extract or fraction thereof as an active ingredient.

In addition, the present invention relates to a method for producing Sangeretia The present invention provides a health functional food composition for preventing or ameliorating a cancer containing an extract or fraction thereof as an active ingredient.

The inventive Sangeretia thea extract and its solvent fractions reduce the cell survival rate of breast cancer cells, induce cell suicide, and can secure the stability from a plant. Therefore, the functional food and medicines for prevention, improvement or treatment of breast cancer . ≪ / RTI >

1 is a homology tree of the present invention (Sageretia when the extracts and their fractions were treated with AGS (gastric cancer cells), A549 (lung cancer cells), MDA-MB-231 (breast cancer cells), SNU-1 (gastric cancer cells) and SNU-16 Survival rate. (B) is the hexane fraction of MB, (C) is the chloroform fraction of MB, (D) is the ethyl acetate fraction of MB, (E) is the butanol fraction of MB and (F) is the water fraction of MB. * Means that there is a statistical difference in comparison with the control group in which the homologous methanol extract and its fractions are not treated, which means that the p value is less than 0.05.
Fig. 2 is a photograph of the Sangeretia thea) extract and fluorescence micrograph of when the fractions thereof were treated in the MDA-MB-231 (breast cancer cells), cell death (apoptosis) The MB-231 Cells (A), a graph (B) measure the apoptotic cells And flow cytometry analysis (C), which is measured through adenine V / PI (propidium iodide) staining. (B) * means that there is a statistical difference in comparison with the control (control) not treated with the homologous methanol extract and its fractions, meaning that the p value is less than 0.05. Control is the negative control without any treatment, MB is the methanol extract of the mixture of leaves and branches of the same tree, HF is the hexane fraction of MB, CF is the chloroform fraction of MB, EF is the ethyl acetate fraction of MB, and BF Is the butanol fraction of MB and WF is the water fraction of MB.

According to an aspect of the invention, the invention is homologous tree (Sageretia The present invention provides a pharmaceutical composition for preventing or treating cancer containing an extract or fraction thereof as an active ingredient.

In the pharmaceutical composition for preventing or treating cancer according to the present invention, the above-mentioned Sambrook extract may be obtained by extracting a mixture of leaves and branches of a Sambrook tree, preferably 1 to 10: 10-1 But the present invention is not limited thereto.

In addition, the extract of the above-mentioned Sambrook may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, preferably methanol or ethanol, more preferably methanol But the present invention is not limited thereto.

In addition, the fraction may be a hexane fraction, a chloroform fraction, an ethyl acetate fraction, a butanol fraction, or a water fraction of a methanol extract of a homologue, but is not limited thereto.

Also. The above-mentioned methanol extract of the same or fractions thereof may be used in combination with 2-Hydroxy-3-methyl-4-pyrone, 3,3,5,5-tetramethyl- (3,3,5,5-Tetramethyl-1,2-cyclopentanedione), 3,5-dihydroxy-6-methyl-2,3-dihydro-4H- Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one, isonicotinic acid-β-D2, Dodecanol, Acetoveratrone, 2,5-dimethoxy-4-methylbenzaldehyde, tetradecyl acrylate, neophytadiene, 6,10,14-trimethyl-2- But are not limited to, 6,10,14-trimethyl-2-pentadecanone, palmitic acid, methyl palmitoleate, dodecyl 2-nitrophenyl ether, But are not limited to, margaric acid, galactopyranoside, isobutyl caprylate, Methyl linoleate, Methyl oleate, Ethyl linoleate, Methyl stearate, Olealdehyde, 9,12,15-octadecatriene-1 -9-octadecatrien-1-ol, stearic acid, 2-palmitoylglycerol, 6-hexyltetrahydro-2H-pyran- Hexyltetrahydro-2H-pyran-2-one, 2,6-dimethyl-N- (2-methyl- alpha-phenylbenzyl) aniline, 3-nitropthalic acid, 1-propene, 3- (2-cyclopentenyl) -2-methyl-1,1- 2-methyl-1,1-diphenyl-, Clionasterol, Dimethyl 3,3'-Thiodipropionate, Friedelin, (Oleamide), 1,1,6,6-tetramethylspiro [4.4] nonan-2-one, 1,1,6,6-tetramethylspiro [ , 2,3,5-tetra (1, 3, 5-Tetraisopropylcyclohexane), Solanesol, octadecyl 3,5-di-t-butyl-4-hydroxyhydrocinnamate -butyl-4-hydroxyhydrocinnamate, dilauryl thiodipropionate, icosamethylcyclodecasiloxane, 1-hydroxymethyl-4,5,7-tri-T- 4-epo, 2-benzyloxy-2-methylhept-3-ene -ene, (24R, 25S) -apristeryl acetate ((24R, 25S) -aplysterylacetate), ethyltrimethyllead, silicone grease, 1,1,3,3,5,5- (1,1,3,3,5,5-hexamethyl-cyclohexasiloxane), Chlorthiophos oxygen analog, (5Z, 7Z) -1,2,3,4,6a , 6b, 9,10,11,12,12a, 12b-dodecahydrocycloocta [3,4] cyclobuta [1,2] [8] 4,6a, 6b, 9, 10 , 11,12,12a, 12b-Dodecahydrocycloocta [3,4] cyclobuta [1,2] [8] annulene, Norolean-12-ene, N- [3- (2-methoxycarbonylphenoxy) propyl] 3,4-dihydroxyphenylacetamide, Hop-22 (29) - ene (Hop-22 (29) -ene, and 2,2-dibenzylethanol, but the present invention is not limited thereto.

In addition, the cancer may be breast cancer, but is not limited thereto.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions.

The pharmaceutical dosage forms of the compositions according to the present invention may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable set.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method . Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition including the above-mentioned extracts of the same or a fraction thereof include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, Various compounds including gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And mixtures thereof. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose , Gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art.

In addition, the present invention relates to a method for producing Sangeretia The present invention provides a health functional food composition for preventing or ameliorating a cancer containing an extract or fraction thereof as an active ingredient.

The composition may be any one selected from powders, granules, pills, tablets, capsules, candies, syrups and beverages, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The active ingredient may be suitably used depending on its intended use (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in the case of long-term intake intended for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount of more than the above range.

There is no particular limitation on the kind of the health functional food. Examples of the foods to which the health functional food composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention may contain ingredients that are conventionally added. For example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of a drink, a natural carbohydrate or a flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like. Although the ratio of the above-mentioned ingredients is not critical, it is generally selected in the range of 0.01 to 0.1 part by weight based on 100 parts by weight of the health functional food composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Experimental Methods and Materials

1. Experimental material

The leaves and branches of Sageretia thea (Osbeck) MCJohnst. Used in the present invention were collected in November 2013 in Seogwangri, Seogwipo, Seogwipo city, Jeju island, and stored frozen at -70 ℃, and used as a sample in the following examples.

2. Extracts and their Fraction  Produce

The extract was prepared by adding 3.75 L of 80% (v / v) methanol and 80% (v / v) ethanol to the mixture of the leaves and branches of the same tree at a weight ratio of 1: Ultrasonic extraction was repeated three times. The leaf extracts and the mixture extracts of the leaves and branches were filtered through a filter of Whatman No. 2, respectively, and then concentrated under reduced pressure at 40 ° C using a rotary vacuum evaporator. Dried at -80 to -60 ° C for 48 hours using a freeze-drier (PVTFD10R, Ilsin, Korea) to prepare a powder, which was then stored frozen and used as an analytical sample in the following examples. The fraction with butanol (n -butanol), and water (water) - in the leaf and of the mixture of homologous wood methanol extracts sequentially n - hexane (n -hexane), chloroform (chloroform), ethyl acetate (ethyl acetate), n Fractions were obtained for each solvent.

3. Total polyphenol and flavonoid content

Total polyphenol content was determined by adding 125 μl of sample to 1,375 μl of distilled water, adding 0.5 ml of Folin-Ciocalteu's reagent, adding 1 ml of Na ₂ CO ₃ after 3 minutes, After standing for 30 minutes, absorbance was measured at 700 nm using a spectrophotometer (Sunrise, Tecan, Salzburg, Austria). Total polyphenol content was expressed as gallic acid equivalents (mg GAE / g).

Total flavonoid content was determined by adding 6 ㎕ of 5% NaNO ₂ to the sample of 40 ㎕, reacting for 5 minutes, adding ₁₂ ㎕ of 10% AlCl ₃ , mixing and reacting for 6 minutes, adding 40 ㎕ of 1N NaOH, Sunrise, Tecan). The absorbance was measured at 510 nm. Total flavonoid content was expressed as rutin equivalents (mg RE / g).

4. ABTS  Radical Scatters (radical scavenging activity)

The ABTS solution was prepared by mixing ₇ mM ABTS (2,2'-azinobis (3-ethylbensthiazoline-6-sulfonic acid) and 2.45 mM K ₂ S ₂ O ₈ in a dark place for 16 hours. The OD value was 0.700 ± 0.005 The above-mentioned 900 μl of ABTS solution and 100 μl of sample were mixed and placed in a cuvette. After reacting at room temperature for 2 minutes, the absorbance was measured at 734 nm using a spectrophotometer (UV1800, Shimadzu) - tocopherol was used as a positive control.

5. Alkyl Radical  Scavenging ability Alkyl radical scavenging activity )

The alkyl radical scavenging activity was determined by adding 20 μl of distilled water, 40 mM AAPH (2-methylpropionamidine) dihydrochloride and 40 mM 4-POBN (alpha- 20 μl of each concentration sample was added sequentially and reacted in a constant temperature water bath at 37 ° C for 30 minutes and then transferred to a capillary tube and measured using an electron spin resonance (ESR) spectrometer (JES-FA200, JEOL, Tokyo, Japan) Respectively. The measurement conditions were a magnetic field of 336,000 mT, a power of 7 mW, a sweep time of 30 seconds, a sweep width of 10 mT, a frequency of 9.43 GHz, a modulation width of 0.2 mT, Amplitude 500 and time constant 0.03 seconds.

6. DPPH  Radical Scatters (radical scavenging activity)

DPPH radical scavenging activity was measured according to the method of Hunting et al. (Nanjo F, et al., 1996, Free Radic Bio Med., 21, 895-902). 30 μl of 60 μM DPPH dissolved in ethanol and 30 μl of the prepared sample were mixed, stirred for 10 seconds, reacted at room temperature for 2 minutes, transferred to a capillary tube, and measured using an ESR spectrometer. The measurement conditions were a magnetic field of 336,000 mT, a power of 5 mW, a sweep time of 30 seconds, a sweep width of 10 mT, a frequency of 9.43 GHz, a modulation width of 0.8 mT, Amplitude 500 and time constant 0.03 seconds. As a positive control, catechin was used.

7. Cell culture

SNU-1 (gastric cancer cell line) and SNU-16 (gastric cancer cell line) used in this experiment were purchased from the Korean Cell Line Bank. Respectively. Cell lines were obtained by using DMEM (Dulbecco's modified Eagle's medium) supplemented with 10% FBS (fetal bovine serum), 100 units / mL penicillin and 100 μg / mL streptomycin, RPMI and F-12K medium And cultured in a 5% CO ₂ incubator at 37 ° C.

8. Cytotoxicity measurement

The cytotoxicity of the extracts of the present invention and its fractions against cancer cells was measured using MTT assay. AGS, A549 and MDA-MB-231 cell line is 3 × 10 ⁴ cells / mL for SNU-1 and SNU-16 cell line is 5 × 10 ⁴ cells / mL and dispensed to a 96-well plate in 5% CO ₂ 37 ℃ And cultured in an incubator for 16 hours. Samples were treated for each concentration and cultured for 24 hours. After removing the culture medium, MTT solution (5 mg / mL, Ameresco Inc.) was treated for 4 hours and then dissolved in 150 μl of DMSO. Absorbance was measured at 570 nm using a spectrophotometer (Sunrise, Tecan). The cell viability was measured by the absorbance reduction ratio of the sample solution and the no-added sample solution.

9. Observing cell morphology

Cell morphology was determined by preparing MDA-MB-231 cell line at 6 × 10 ⁴ cells / mL in a 6-well plate, culturing the samples for 24 hours, treating 10 μM Hoechst 33342 And then observed using a fluorescence microscope (Olympus, Essex, UK).

10. Annexin  V ( Annexin  V) measurement

The presence or absence of the induction of apoptosis of cancer cells when treated with the extracts of the present invention and the fractions thereof was evaluated by using anexin V-FITC Apoptosis Detection Kit-1 in the cells treated with apoptosis ). The cells were treated with MDA-MB-231 cell line for 24 hours, washed with PBS, stained with binding buffer (Annexin V) and PI (propidium iodide) for 15 minutes at room temperature, Cells were measured for annexin V positive cells expressing apoptotic cells using a FACS caliber flow cytometer (BD biosciences).

11. Gas Chromatography-Mass Spectrometry GC -MS)

Gas chromatographic analysis was performed using Shimatsu GC-MS (Model QP-2010, Shimadzu Co., Japan) / EI (electron impact) mode. The ionization voltage was 70 eV and the injector temperature and the interface temperature were 250 ° C and 290 ° C, respectively. Rtx-5MS was used as a GC column (30 m length, 0.25 mm inner diameter and 0.25 μM film thickness). The oven temperature was raised at 3 ° C per minute to 200 ° C after staying at 40 ° C for 2 minutes, then stayed for 5 minutes and then set to rise again to 250 ° C at 5 ° C per minute. The carrier gas helium was injected at a pressure of 57.4 kPa at a flow rate of 1 ml per minute at a 1:10 split ratio and the mass range was 40 to 500 amu. MS spectral data were compared within the WILEY9 and NIST05 libraries.

Example 1. homologous trees (Sageretia antioxidant activity of theaa extract

Since the leaves of Sambrook tree are very small and difficult to utilize, leaf and branches were extracted together and their antioxidative activities were compared with leaf extract alone. As a result, as shown in Table 1, the methanol extract (MB), the ethanol extract (EB), the methanol extract (ML) and the homologous The extraction yield of ethanol extract (EL) of tree leaves was highest at 20.43%, and EB, EL and ML were measured as 19.33%, 15.44% and 9.69%, respectively. The total polyphenol content and total flavonoid content of the extracts were 37.2 mg GAE / g and 7.8 mg RE / g, respectively, and the total polyphenol content and total flavonoid content were the highest among the four extracts. ABTS, alkyl (Alkyl), and through radical scavenging activity on DPPH for analysis, the results of comparing the antioxidant activity of the extract, to the MB, as shown in Table 2 IC ₅₀ Values were 55.4㎍ / mL (ABTS radical scavenging activity), IC ₅₀ 37.0㎍ / mL (alkyl radical scavenging activity) and activation of the IC ₅₀ 8.4㎍ / mL (DPPH radical scavenging activity). In conclusion, MB showed higher total polyphenol and total flavonoid content and higher antioxidant activity than EB, and the mixture extracts (MB and EB) of the leaves and branches of the same tree were the only leaf extract ) And the antioxidative activity were superior to those of the extracts. Therefore, the following Example 2 confirmed the antioxidative activity and cytotoxicity of the methanol extract (MB) and its solvent fractions of the leaf and branch mixture of the same tree.

Extraction yield, total polyphenol and total flavonoid content of extract extract EL EB ML MB Extraction yield (%) 15.44 19.33 9.69 20.43 Total polyphenol content (mg GAE / g) 23.9 ± 2.0 28.3 ± 4.3 26.4 ± 0.8 37.2 ± 4.5 Total flavonoid content (mg RE / g) 6.1 ± 0.5 6.9 ± 0.5 6.1 ± 0.8 7.8 ± 0.5

EL: Ethanol extract of Safflower leaves

EB: Ethanol Extract of Safflower Leaf and Branch Mixtures

ML: Methanol Extract of Safflower Leaf

MB: Methanol Extract of Safflower Leaf and Branch Mixtures

The antioxidative activity (IC ₅₀ ㎍ / mL) of the extracts of each part extract EL EB ML MB ABTS radical scavenging ability 84.4 ± 6.6 64.2 ± 2.1 78.1 ± 10.5 55.4 ± 5.1 Alkyl radical scavenging ability 52.5 ± 7.2 52.6 ± 7.6 52.1 ± 11.3 37.0 + - 6.7 DPPH radical scavenging ability 18.2 ± 1.5 14.0 ± 0.9 15.3 ± 1.0 8.4 ± 0.9

IC ₅₀ ㎍ / mL: The half maximal inhibitory concentration on radical scavenging activity.

Example  2. Same tree  Solvent of extract (MB) In the fraction  Antioxidant activity

Successively methanol extract (MB) of the leaf and of the mixture of homologous tree in this embodiment, 2 n - hexane (n -hexane), chloroform (chloroform), ethyl acetate (ethyl acetate), n - butanol (n -butanol) And water to obtain fractions for each solvent. Total polyphenol, total flavonoid content and antioxidant activity were measured. As a result, the highest total polyphenol and total flavonoid contents were shown as 57.9 mg GAE / g and 9.8 mg RE / g in the ethyl acetate fraction (EF) among the solvent fractions as shown in Table 3 below, and n -butanol fraction (BF) has showed a high content of the second to 40.8mg GAE / g and 9.8mg RE / g, followed by methanol extract (MB), the water fraction (WF), chloroform fraction (CF) and n - hexane Fraction (HF). Antioxidant activity was similar to that of total polyphenol and total flavonoid contents. EF and BF were measured by ABTS (IC ₅₀ 35.0 and 46.4 μg / mL), Alkyl (IC ₅₀ 17.8 and 14.6 μg / mL) and DPPH ₅₀ 4.8 and 7.0 / / mL) showed the highest activity in the radical scavenging assay (Table 4).

Extraction yield, total polyphenol and total flavonoid content of Safflower extract and its fractions extract MB HF CF EF BF WF Extraction yield (%) 20.43 1.75 3.83 7.74 14.56 55.74 Total polyphenol content
(mg GAE / g) 37.2 ± 4.5 9.3 ± 6.4 11.9 ± 3.1 57.9 ± 6.4 40.8 ± 6.1 35.8 ± 7.6 Total flavonoid content
(mg RE / g) 7.8 ± 0.5 - 4.5 ± 0.4 9.8 ± 0.6 9.8 ± 0.5 7.6 ± 1.0

MB: methanol extracts of the leaves and branches of the alien tree; HF: n -hexane fraction of MB; CF: chloroform fraction of MB; EF: ethyl acetate fraction of MB; BF: n - butanol fraction of MB and WF: water fraction of MB.

The antioxidative activity (IC ₅₀ ㎍ / mL) of Safflower extract and its fractions extract MB HF CF EF BF WF ABTS radical scavenging ability 55.4 ± 5.1 > 100 > 100 35.0 ± 3.8 46.4 ± 2.1 59.6 ± 4.6 Alkyl radical scavenging ability 37.0 + - 6.7 > 50 > 50 17.8 ± 4.7 14.6 ± 4.6 43.0 ± 2.3 DPPH radical scavenging ability 8.4 ± 0.9 > 10 > 10 4.8 ± 1.2 7.0 ± 0.7 8.1 ± 0.2

Example  3. Same tree  Solvent of extract (MB) Fraction  Cell death (MDA-MB-231) in breast cancer cell lines Apoptosis ) Judo

In Example 3, fractions of solvent extracts of methanol extract (MB) of leaf and branch mixture of the same tree were treated with AGS (gastric cancer cell), A549 (lung cancer cell), MDA-MB-231 (breast cancer cell), SNU- Cells) and SNU-16 (gastric cancer cells), cytotoxicity of the cancer cells was measured by MTT assay. As a result, as shown in FIG. 1, the MB and its solvent fractions showed cytotoxicity in a concentration-dependent manner, especially MDA-MB-231 (breast cancer cells), and the cell survival rate at 100 μg / mL was 42.72% ), 52.08% (EF), 48.96% (BF), 46.81% (WF), 78.38% (HF) and 72.89% (CF). To determine whether the cytotoxicity of the solvent fraction of MB was due to apoptosis, DNA fragmentation (cleavage) and degree of condensation were confirmed by Hoechst 33342 staining. As a result, MB (15.6%) and EF (12.3%) were treated with MDA-MB-231 (breast cancer cells) , And BF (19.4%) and WF (9.0%), respectively, but not in HF and CF. In addition, Annexin V / PI staining showed that the apoptotic cell annexin V-positive cells showed 52.2% (BF), 51.7% (BF) MB), 44.5% (EF), 39.8% (HF), 37.5% (WF) and 9.2% (CF), respectively. In conclusion, it was confirmed that the treatment of MB fraction with solvent resulted in apoptosis in MDA-MB-231 (breast cancer cells).

Example  4. GC / MS Same tree  Of extract (MB) Fraction  Component analysis

In Example 4, the methanol extract (MB) and the n -hexane fraction (HF), the chloroform fraction (CF), the ethyl acetate fraction (EF), the n -butanol fraction HF, CF, EF, BF and WF were analyzed by GC / MS for water fraction (WF) and water fraction (WF). As a result, a total of 53 compounds could be detected as shown in Table 5 below.

Of Same Tree Extract (MB) Analysis of components of fractions No. RT ¹⁾ (min) ingredient domain(%) MB HF CF EF BF WF One 10.489 2-Hydroxy-3-methyl-4-pyrone 1.43 ND ²⁾ ND ND ND ND 2 11.377 3,3,5,5-Tetramethyl-1,2-cyclopentanedione 5.73 ND ND ND ND ND 3 12.362 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one 4.29 ND ND ND ND ND 4 13.584 Isonicotinic acid -? - D2 1.83 ND ND ND ND ND 5 17.183 Dodecanol ND ND 2.31 ND ND 7.77 6 20.858 Acetoveratrone ND ND ND ND ND 1.78 7 24.010 2,5-Dimethoxy-4-methylbenzaldehyde 1.81 ND ND ND ND ND 8 25.385 Tetradecyl acrylate ND 3.07 13.49 1.82 4.52 18.95 9 30.062 Neophytadiene 1.24 0.24 ND ND ND ND 10 30.790 6,10,14-Trimethyl-2-pentadecanone ND 1.59 0.26 ND ND ND 11 32.676 Palmitic acid 4.16 ND ND ND ND ND 12 33.430 Methyl palmitoleate ND ND 1.09 ND ND ND 13 33.561 Methyl palmitate ND 2.88 8.59 1.14 3.58 1.83 14 34.854 Pentadecanoic acid ND ND 0.91 10.98 23.43 5.81 15 36.473 Dodecyl 2-nitrophenyl ether ND 1.11 7.91 3.02 11.12 20.89 16 38.046 Margaric acid ND ND ND 0.33 1.27 ND 17 38.468 Galactopyranoside 2.56 ND ND ND ND ND 18 38.575 Isobutyl caprylate 1.14 ND ND ND ND ND 19 39.039 Methyl linoleate 3.48 2.15 2.19 ND ND ND 20 39.219 Methyl oleate ND ND ND 0.96 3.55 ND 21 39.290 Ethyl linoleate 0.70 5.50 13.32 0.16 ND ND 22 39.701 Phytol 5.84 15.23 1.47 ND ND ND 23 40.021 Methyl stearate ND 1.08 3.38 0.31 ND 0.78 24 40.491 Olealdehyde ND ND ND 2.06 ND ND 25 40.523 9,12,15-octadecatrien-1-ol ND ND ND ND 10.19 ND 26 41.173 Stearic acid ND ND ND 1.78 6.35 1.48 27 42.436 2-Palmitoylglycerol 1.61 ND ND ND ND ND 28 46.797 6-Hexyltetrahydro-2H-pyran-2-one ND 3.28 ND ND ND ND 29 52.066 2,6-dimethyl-N- (2-methyl-.alpha.-phenylbenzyl) aniline ND 1.93 ND ND ND ND 30 52.070 3-Nitrophthalic acid ND ND ND 1.85 ND ND 31 52.429 1-Propene, 3- (2-cyclopentenyl) -2-methyl-1,1-diphenyl- ND 1.71 ND ND ND ND 32 52.577 Clionesterol 3.18 ND ND ND ND 0.56 33 52.673 Dimethyl 3,3'-Thiodipropionate ND ND 1.02 ND ND 3.52 34 57.018 Friedelin ND ND 22.63 ND ND ND 35 58.139 Oleamide 1.24 1.08 1.96 0.62 3.94 6.96 36 59.004 1,1,6,6-Tetramethylspiro [4.4] nonan-2-one ND ND ND ND 1.04 ND 37 59.101 Lycopersene ND 2.46 ND ND ND ND 38 59.732 [delta] -Tocopherol ND 3.91 ND ND ND ND 39 60.943 1,2,3,5-Tetraisopropylcyclohexane ND 1.01 ND ND ND ND 40 61.223 Solanesol ND 1.02 ND ND ND ND 41 61.285 Octadecyl 3,5-di-t-butyl-4-hydroxyhydrocinnamate ND ND 6.63 0.44 ND 0.52 42 61.560 Dilauryl thiodipropionate 35.40 ND ND 1.68 16.60 25.50 43 61.660 Icosamethylcyclodecasiloxane ND ND ND 3.64 2.51 ND 44 61.875 1-Hydroxymethyl-4,5,7-tri-T-butyl-1,4-epo ND ND 3.38 ND ND ND 45 63.541 2-benzyloxy-2-methylhept-3-ene ND 1.00 ND ND ND ND 46 64.280 (24R, 25S) -aplysterylacetate ND ND ND 2.17 ND ND 47 64.818 α-tocopherol-β-D-mannoside 14.64 13.69 1.12 ND 0.99 ND 48 65.575 Ethyltrimethyllead ND ND ND ND 1.59 ND 49 65.925 Silicone grease ND ND 0.27 ND 1.07 ND 50 65.985 1,1,3,3,5,5-hexamethyl-cyclohexasiloxane ND ND ND ND 1.37 ND 51 66.040 Chlorthophos oxygen analog ND ND ND ND 1.60 ND 52 66.140 (5Z, 7Z) -1,2,3,4,6a, 6b, 9,10,11,12,12a, 12b-Dodecahydrocycloocta [3,4] cyclobuta [1,2] [8] annulene ND ND ND ND 1.44 ND 53 67.906 β-sitosterol ND 9.52 2.54 59.49 3.82 ND 54 68.688 Norolean-12-ene ND 2.27 ND ND ND ND 55 69.080 N- [3- (2-methoxycarbonylphenoxy) propyl] 3,4-dihydroxyphenylacetamide ND ND ND 7.32 ND ND 56 69.185 Hop-22 (29) ND 1.01 ND ND ND ND 57 69.272 2,2-Dibenzylethanol ND ND 1.10 ND ND ND 58 69.572 Lupeol acetate 1.28 7.66 ND ND ND ND gun 91.56 84.40 95.57 99.77 99.98 96.35

¹⁾ RT: Retention time

²⁾ ND: Not detected

Claims (10)

A pharmaceutical composition for the prevention or treatment of breast cancer, which comprises a butanol fraction of Sageretia thea methanol extract as an active ingredient. The pharmaceutical composition for preventing or treating breast cancer according to claim 1, wherein the methanol extract of Sageretia thea is a mixture of leaves and branches of the same tree. delete delete delete delete The pharmaceutical composition for preventing or treating breast cancer according to claim 1, further comprising a pharmaceutically acceptable carrier, excipient or diluent in addition to the butanol fraction of the same methanol extract. The pharmaceutical composition for preventing or treating breast cancer according to claim 1, wherein the composition is prepared from any one selected from the group consisting of capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols. A health functional food composition for preventing or ameliorating breast cancer, which contains butanol fraction of methanol extract of Sageretia thea as an active ingredient. The health functional food composition according to claim 9, wherein the composition is prepared from one of powder, granule, ring, tablet, capsule, candy, syrup and beverage.

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