KR102471017B1 - Anticancer composition comprising Lonicera subsessilis extract - Google Patents

Abstract

The present invention relates to an anti-cancer composition containing an extract of japonica japonica, and a pharmaceutical composition or a health functional food containing an extract of a japonica japonica has an excellent effect on killing lung cancer cells, thereby preventing various cancer-related diseases including lung cancer. It can be easily used for improvement, treatment, etc.

Description

Anticancer composition comprising Lonicera subsessilis extract}

The present invention relates to a composition for anti-cancer containing an extract of B. chinensis.

Malignant tumor, or cancer, the worst disease threatening human health, is the second most common cause of death in Western societies after cardiovascular disease. In modern society, lung cancer due to an aging population, an increase in the smoking population, and air pollution are significantly increasing. Intake of a high-fat diet, increase in environmental pollutants, and increase in alcohol consumption due to westernization of diet lead to lung cancer, colon cancer, breast cancer, and prostate cancer. etc. are continuously increasing. Therefore, there is an urgent need for the creation of anti-cancer substances that can contribute to the improvement of human health, the improvement of healthy quality of life, and the promotion of human health by enabling early prevention and treatment of cancer. Drugs developed for cancer treatment have low specificity for cancer cells because they are non-specific and act throughout the body along the bloodstream following systemic injection or oral administration. Therefore, there are many systemic side effects according to drug administration, and there are many disadvantages in that side effects appear more than surgery or radiation therapy. As the cancer treatment period using existing anticancer drugs is administered 3 to 5 days a week at intervals of 3 to 4 weeks for 6 months to 1 year, many cases of stopping treatment due to various side effects of anticancer drugs occur.

Existing chemical therapies that exert their medicinal effects by directly attacking cancer cells have very strong side effects and have limited clinical use despite the disadvantages of non-specificity. Accordingly, many inventors are actively conducting various studies to find natural materials having anticancer activity.

Lonicera subsessilis Rehder is a deciduous shrub of the dicotyledonous plywood group, the honeysuckle family, and grows in deep mountain forests. It is about 2m in height. The small branch is purplish brown, and the eye scales remain under the new branch, and the inside of the branch is white. The leaves are opposite, egg-shaped or egg-shaped oval, with flat edges and hairless. Flowers bloom in June, are white or light yellow, and hang on the axils of new branches. The peduncle is 4-5mm long, and the bracts are shorter than the parcels, and there are preemptive dots, and the parcels are combined to form a dish. The upper lip of the corolla is divided into 3 parts, which is longer than the tube part. There are 4 stamens, similar to the length of the corolla branch, and the style is rather long. Two ovaries are almost combined, and the fruit is round and ripens in red in August-September. As a Korean endemic species, it is distributed in Chungcheongbuk-do, Pyongannam-do, and Hamgyeongbuk-do. The pharmacological effects are not yet well known.

Accordingly, the inventors of the present invention completed the present invention by confirming that the extract of the japonica japonica has an excellent killing effect on lung cancer cells while studying various physiological activities of the japonica japonica.

Republic of Korea Patent Registration No. 10-1833036 (Title of Invention: Composition for treatment or prevention of liver cancer or kidney cancer containing gold and silver flower extract, Applicant: Institute for Integrated Medical Promotion) Republic of Korea Patent Publication No. 10-1994-0001893 (Title of Invention: Anti-inflammatory, anti-allergic and anti-rheumatic agent of biflavonoid obtained from Lonicera japonica, Applicant: Kang Sam-shik and 3 others)

Paramasivan Poornima (2014). Doxorubicin induced apoptosis was potentiated by neferine in human lung adenocarcima, A549 cells, Food and Chemical Toxicology, 68, 87-98. Maria Davoren (2007). In vitro toxicity evaluation of single walled carbon nanotubes on human A549 lung cells, Toxicology in Vitro, 21, 438-448.

An object of the present invention is to provide a composition for anti-cancer containing an extract of japonica japonica.

The present invention relates to an anti-cancer composition containing an extract of Lonicera subsessilis Rehder.

The extract of the firewood tree can be extracted by using water, C1-C4 alcohol, or a mixture thereof as a solvent. The solvent may be an alcohol aqueous solution, preferably a 50 to 80% (v/v) ethanol aqueous solution.

The cheonggoebul tree extract may be a leaf extract of cheonggoe fire tree.

The present invention can thus provide a composition for preventing or treating cancer containing the extract of the chrysanthemum tree.

The cancer may be a disease selected from lung cancer, colon cancer, cervical cancer, pituitary gland cancer, brain tumor, stomach cancer, liver cancer, breast cancer, pancreatic cancer, uterine cancer, leukemia, esophageal cancer, prostate cancer, bladder cancer and ovarian cancer, preferably lung cancer. can

Therefore, the present invention may relate to a health functional food for preventing or improving cancer containing an extract of chrysanthemum japonica.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for anti-cancer containing an extract of B. chinensis. In addition, the present invention relates to a composition for preventing or treating cancer containing the extract of the fire tree, wherein the extract of the fire tree can be extracted from water, C1 to C4 alcohol or a mixture thereof as a solvent. And, the C1 to C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.

Water, C1-C4 alcohol, or a mixed solution thereof used in the preparation of the Bronx tree extract may be used in an amount of 1 to 50 times the volume (1 to 50 ℓ based on 1 kg) based on the weight of the Bul tree extract, preferably 5 ~50 times the volume can be used. The extraction conditions for the extract of the chrysanthemum tree extract may be 1 minute to 48 hours at 20 to 100 ° C, and preferably 2 to 24 hours at 20 to 40 ° C. The above process may be repeated from 1 to 4 times. Sonication may be performed during the extraction time.

In addition, as a conventional method in the art, after dissolving the extract of water, C1-C4 alcohol, or a mixture solution thereof of the green locust tree in water, selected from the group consisting of n-hexane, methylene chloride, ethyl acetate and n-butanol It may be prepared into fractions by further fractionation using one or more solvents.

More specifically, water is added to and suspended in the extract of the fire tree obtained by extracting and concentrating the tree of the tree with water, C1 ~ C4 alcohol, or a mixed solvent thereof, preferably 1 to 1000 of the weight of the tree of the tree. After adding and suspending the suspension by adding 10 to 50 times more water, more preferably 5 to 500 times, and most preferably 10 to 50 times, a solvent selected from the group consisting of hexane, methylene chloride, ethyl acetate and n-butanol is added to the suspension. It can be manufactured from the sapwood fraction obtained by adding

More preferably, the buckwheat tree fraction is obtained by suspending the buckwheat tree extract obtained by extracting and concentrating the buckwheat tree extract with water, C1-C4 alcohol, or a mixed solvent thereof in water and then mixing it with n-hexane. -The concentrate of the hexane layer, the concentrate of the methylene chloride layer obtained by mixing methylene chloride with the remaining water layer after removing the n-hexane layer, or the ethyl acetate obtained by mixing ethyl acetate with the remaining water layer after removing the methylene chloride layer It may be a concentrate of a layer, a concentrate of an n-butanol layer obtained by mixing n-butanol with an aqueous layer remaining after removing the ethyl acetate layer, or a concentrate of an aqueous layer remaining after removing the butanol layer.

As a device for extracting the extract or a fraction thereof, a conventional extraction device, an ultrasonic pulverization extractor, or a fractionator may be used. The extract or fraction prepared in this way may be dried under reduced pressure or lyophilized to remove the solvent. In addition, the extracts or fractions of the japonica japonica may be used after being purified using column chromatography.

The extract of the sagebrush tree extract is obtained by a known method used for separation and extraction of plant components, such as extraction with an organic solvent (alcohol, ether, acetone, etc.), distribution of hexane and water, and column chromatography, according to a conventional method. Alternatively, it may be used after fractionation or purification using a suitably combined method.

The chromatography is silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography), thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

In addition, the present invention provides a pharmaceutical composition for anti-cancer, a pharmaceutical composition for preventing or treating cancer containing the extract or a fraction thereof. The extract or a fraction thereof may be added in an amount of 0.001 to 30% by weight in the pharmaceutical composition of the present invention.

The pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, and cellulose. , methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, for example, starch, calcium carbonate, sucrose or It is prepared by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin fat, glycerogeratin and the like may be used.

The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. Determination of dosage based on these factors is within the level of those skilled in the art, and generally dosages range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 1 mg/kg/day to 500 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, and humans through various routes. All modes of administration are contemplated, eg oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection. Since the extract or fraction of the present invention has little toxicity and side effects, it is a drug that can be safely used even when taken for a long period of time for preventive purposes.

In addition, the present invention provides a health functional food for anti-cancer, a health functional food for preventing or improving cancer, containing a food-acceptable food supplement additive of the extract or fraction of the eucalyptus fir tree. The cheonggoebul tree extract may be added to the health functional food of the present invention in an amount of 0.001 to 100% by weight. The health functional food of the present invention includes forms such as tablets, capsules, pills or liquids, and foods to which the extract of the present invention can be added include, for example, various drinks, meat, sausages, bread, candy, Snacks, noodles, ice cream, dairy products, soups, ionic beverages, soft drinks, alcoholic beverages, chewing gum, tea, and vitamin complexes.

The present invention relates to an anti-cancer composition containing an extract of japonica japonica, and a pharmaceutical composition or a health functional food containing an extract of a japonica japonica has an excellent effect on killing lung cancer cells, thereby preventing various cancer-related diseases including lung cancer. It can be easily used for improvement, treatment, etc.

1 is a graph showing the apoptosis effect of lung cancer cell extract of cheongoebul tree extract.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to sufficiently convey the spirit of the invention to those skilled in the art, so that the disclosure herein will be thorough and complete.

<Example 1. Preparation of extracts of japonica japonica>

After putting 1 kg of Lonicera subsessilis Rehder leaves in 10 L of 70% (v/v) ethanol aqueous solution and submerging the sample sufficiently, extraction was performed by applying ultrasonic waves while stirring at room temperature (25 ° C) for 24 hours. . This process was repeated three times to sufficiently extract useful components. After first filtering using sterile gauze, the filtrate was filtered again using filter paper (Whatman No. 2; Whatman Ltd., Maidstone, Kent, UK) to filter out pulverized samples. Then, the residue was removed using a filter paper to obtain an extract filtrate. This operation was repeated three times to combine all the filtrates, and then concentrated under reduced pressure using a rotary evaporator to obtain an extract, and the remaining solvent was removed as much as possible by lyophilization. Lyophilized samples were stored at -20 ° C until use in experiments.

<Example 2. Confirmation of Lung Cancer Cell Apoptosis Effect of Lung Tree Extract>

Confirmation of the killing effect of lung cancer cells was confirmed by MTT assay. After adding 1 x 10 ⁴ cells/100 μl of A549 cells (human lung carcinoma cell line, sold by Korea Cell Line Bank) per well to a 96 well plate, incubation was performed in an incubator for 24 h. After removing the media in the plate, after processing 100 μl of sample and positive control (adriamycin) at the corresponding concentration, incubation in an incubator for 48 hours, removing the media in the plate, processing 100 μl of 0.2 mg/ml MTT, and then 3- It was incubated in an incubator for 4 h. Next, after removing the MTT solution in the plate, adding DMSO (dimethyl sulfoxide), shaking for 30 minutes, and measuring at 545 nm using a microplate reader (Tecan, Switzerland). At this time, the control group was a solvent control group, and the positive control was adriamycin 2 μM, and each sample was treated with the leaf extract obtained in Example 1 for each concentration. Evaluation of cytotoxicity (anti-cancer activity) was expressed as a relative ratio when the survival rate of the control group was 100%.

The results for this are shown in FIG. 1, and referring to the results of FIG. 1, it is confirmed that the extract of Bulgogi japonica effectively kills A549 cells, which are lung cancer cell lines.

<Example 3. Lonicera Comparison of lung cancer cell killing effect of genus plant extracts>

Next, the apoptotic effect on the lung cancer cell line A549dp was measured at a concentration of 100 ppm by the method of Example 2 of leaf extracts of plants of the genus Lonicera .

target plant A549 cell viability Big tree ( Lonicera maackii ) 101.6 ± 3.4%

Referring to the results in Table 1, the extract of the Lonicera genus, another species of plant, has no higher anticancer activity than the extract of the Lonicera genus. You can check.

<Formulation Example 1. Pharmaceutical formulation - Preparation of tablets>

200 g of the extract of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. After adding 10% gelatin solution to this mixture, it was pulverized and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

<Formulation Example 2. Food Manufacturing>

Formulation Example 2-1. Preparation of seasoning for cooking

A health-promoting cooking seasoning was prepared by adding 1% by weight of the extract of Bulgogi japonica of the present invention to the cooking seasoning.

Formulation example 2-2. Manufacture of Flour Food

Bread, cakes, cookies, crackers, and noodles were prepared using the mixture by adding 0.1% by weight of the extract of the green tree of the present invention to wheat flour, thereby preparing health-enhancing foods.

Formulation Example 2-3. Preparation of soups and gravies

Health-promoting soup and broth were prepared by adding 0.1% by weight of the extract of the green tree of the present invention to soup and broth.

Formulation Example 2-4. Manufacture of dairy products

0.1% by weight of the extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 2-5. vegetable juice manufacturing

Vegetable juice for health promotion was prepared by adding 0.5 g of the extract of the present invention to 1,000 ml of tomato juice or carrot juice.

Formulation Example 2-6. fruit juice manufacturing

Fruit juice for health promotion was prepared by adding 0.1 g of the extract of the present invention to 1,000 ml of apple juice or grape juice.

Claims (8)

An anti-lung cancer composition comprising a 50-80% (v/v) ethanol aqueous solution extract of Lonicera subsessilis Rehder. delete A composition for preventing or treating lung cancer, comprising a 50-80% (v/v) aqueous ethanol extract of Lonicera subsessilis Rehder. delete delete Health functional food for preventing or improving lung cancer, characterized in that it contains an extract of 50 to 80% (v / v) ethanol aqueous solution of Lonicera subsessilis Rehder. delete delete

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