KR101556443B1 - A composition for anti-cancer comprising extract of Pinus Koraiensis SIEB - Google Patents

Abstract

The present invention provides an anticancer composition containing a pine tree leaf extract. The pine leaf extract of the present invention inhibits the proliferation of cancer cells and exhibits an anti-cancer effect.

Description

[0001] The present invention relates to an anticancer composition containing Pinus koraiensis SIEB

The present invention relates to an anticancer composition containing a natural product extract. Specifically, the present invention relates to an anticancer composition containing a pine tree leaf extract.

Common treatments for cancer include surgery, chemotherapy, and radiation therapy. Surgery is divided into diagnostic surgery, curative surgery, prophylactic surgery, and palliative surgery depending on the purpose. Diagnostic surgery is performed to know the classification and type of tumor. Curative surgery is a method of treating early stage cancer by removing both the lymph nodes surrounding the tumor and the primary lesion. Prophylactic surgery is a method of helping cancer prevention by removing some polyps, previously known as pre-cancerous lesions, and palliative surgery is a way of alleviating the symptoms of cancer by reducing the size of the tumor. Although the growth and differentiation of normal cells in our body is strictly regulated, cancer cells exhibit abnormality patterns that show abnormalities in cell growth regulation and proliferate continuously. Therefore, suppression of cancer cell proliferation is important, and chemotherapy is one of them. It is used for the purpose of treatment of cancer which destroys cancer cells and does not recur, and it is used for the purpose of prolonging the life by inhibiting cancer growth or cancer cell spread in cancer which is not curable. The drugs to be used include alkylating drugs and antimetabolites. The alkylating drug binds directly to DNA, destroying the DNA molecule itself and damaging the same helix or double helix structure, inhibiting the growth and division of cancer cells. Typical drugs include platinum compounds such as cisplatin and carboplatin. Nitrogen mustard drugs include mechlorethamine (nitrogen mustard), cyclophosphamide ). Since the antimetabolite has a structure similar to the metabolites required for DNA replication in normal cells, it is competitive with purine and pyrimidine biosynthetic enzymes and interferes with its action. . Representative examples include pyroridine derivatives such as fluorouracil (5-FU), capecitabine and cytarabine. Examples of purine derivatives include mercaptopurine, 6-MP). Radiation therapy is a method of destroying cancer cells using radiation, sometimes used to alleviate symptoms when the treatment is no longer possible.

However, cancer treatment methods effectively inhibit cancer growth and cause side effects. In the case of surgery, side effects such as pulmonary complications, anastomotic leakage (intestinal lumen) leakage, intestinal obstruction, dysuria, and sexual dysfunction may occur. In cancer chemotherapy, the toxicity of the drug is spread not only in cancer cells but also in normal cells, resulting in some complications. Also, there are gastrointestinal disorders and enteritis, leukocytes and thrombocytopenia, immunosuppression, skin pruritus and hair loss, nervous system toxicity, kidney and hepatotoxicity. In radiation therapy, because of the radiation dose to some tissues as well as normal tissues, the wounds may not heal well, or skin necrosis, myelosuppression, inflammation of the bone marrow, and bleeding may be complications.

Therefore, there is a growing interest in extracts of natural materials having the above-mentioned side effects and anti-cancer effects, and development of safe and efficacious natural drugs is required.

On the other hand, jatip (or also known as 'pine leaf') is white pine (Pinus koraiensis SIEB). It is known that the pine cones contain various essential components such as terpene, phenolics and tannin, and chlorophyll, inorganic and organic components and vitamins. have.

However, the pine needle leaf has been reported by the present inventors to be effective in treating diabetes (Korean Patent Laid-Open No. 10-2010-0115598) and obesity-suppressing activity (Korean Patent Laid-Open Publication No. 10-2012-0134770) Antitumor activity has not been reported.

Accordingly, the inventors of the present invention have conducted research to develop anticancer agents using the pine tree leaves, and as a result, they confirmed that the pine tree leaves inhibit the growth of cancer cells and have anticancer activity, thus completing the present invention.

Korean Patent Publication No. 10-2010-0115598 Korean Patent Publication No. 10-2012-0134770

Disclosure of Invention Technical Problem [8] The present invention provides a pharmaceutical composition which can be administered safely as a natural extract, effectively inhibits the growth of cancer cells, and has excellent anticancer effect, by solving the problems of the conventional art as described above.

The present invention also provides a food composition for preventing or ameliorating cancer.

Pharmaceutical composition

The present invention provides a pharmaceutical composition for preventing or treating cancer containing Pine Pinnae Leaf Extract.

The pine leaf extract of the present invention can be obtained by washing, drying, cutting and crushing, extracting, concentrating and aging the pine tree leaves.

In the present invention, the Pine leaf blade extract can be extracted from the pine tree leaves by a conventional method, for example, a water extraction method, a water + alcohol extraction method, an alcohol extraction method, a steam distillation extraction method and the like. It is obvious to a person skilled in the art that the components and compositions contained in the extract may differ depending on the extraction method. In the present invention, pine tree leaf extract obtained through steam distillation extraction method is particularly preferable.

Steam distillation extraction is well known in the art. Specifically, the ratio of the pine needle blade to the purified water for performing the steam distillation extraction is 1: 1 to 1:10, preferably 1: 1 to 1: 3, more preferably 1: 2 (by weight). Pine tree leaves and purified water in a distillation tank at 70 ° C to 500 ° C, preferably 80 ° C to 150 ° C, more preferably 90 ° C, for about 1 hour to several days, preferably for 3 to 4 hours, Distillation. A condensate obtained by cooling and condensing the distillate thus obtained can be used in the present invention. Further, the thus obtained condensate can be further purified and further filtered by a conventional method in the art, so that a pine cone leaf extract having a higher purity can be used. According to the steam distillation extraction method, the pine tree leaf extract is obtained in the form of clear, colorless transparent oil (distillation-extracting oil) having pine tree incense.

The thus obtained pine leaf extract of the present invention has an effect of effectively inhibiting the growth of cancer cells. Therefore, the Pine leaf blade extract of the present invention can be used for prevention or treatment of cancer.

Specifically, it was confirmed that the pine vine leaf extract of the present invention inhibits the growth of cancer cells, especially colon cancer cells, of gastrointestinal cancer selected from the group consisting of esophageal cancer, stomach cancer, pancreatic cancer, liver cancer, gallbladder cancer, colon cancer and anal cancer. Therefore, the Pine leaf blade extract of the present invention can be used for prevention or treatment of gastrointestinal cancer.

The content of the P. intestinalis leaf extract, which is an active ingredient in the pharmaceutical composition of the present invention, can be appropriately adjusted according to the progress of the cancer, the condition of the patient and the like, for example, 5 to 80% by weight, But is not limited thereto.

The pine needle leaf extract which is contained as an active ingredient in the pharmaceutical composition of the present invention can be formulated together with pharmaceutically or pharmacologically acceptable carriers, excipients or diluents. Remington's Pharmaceutical Science (recent edition) Mack Publishing Company, Easton PA, and the like.

As an embodiment of the present invention, the pharmaceutical composition containing the pine vine leaf extract according to the present invention may be formulated into oral compositions such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, Suppositories, sterile injection solutions, and the like.

Examples of the carrier, excipient and diluent which may be contained herein include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the pharmaceutical composition of the present invention may contain diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and other pharmaceutically acceptable additives which are usually used.

When the composition is formulated as a solid preparation for oral administration, tablets, pills, powders, granules, capsules and the like may be used. In addition to the pine leaf extract, at least one excipient such as starch, calcium carbonate carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used.

When formulated as a liquid preparation for oral administration, suspensions, solutions, emulsions, syrups and the like can be used. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, And the like.

When formulated as a preparation for parenteral administration, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations and suppositories are included. 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, but the present invention is not limited thereto. Examples of suppository bases include, but are not limited to, witepsol, macrogol, tween 61, cacao bean, laurin, glycerogelatin, and the like.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the age, condition and weight of the patient, the degree of progress of the cancer, the type of drug, the route of administration, and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the dose of the composition of the present invention is preferably from 10 mg / kg to 1000 mg / kg, preferably from 100 mg / kg to 1000 mg / kg, on an adult male basis per day, kg.

The administration may be carried out once a day or divided into several doses. However, the scope of the present invention is not limited by the dose and the number of administrations.

The pharmaceutical compositions of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes including, for example, orally, intraperitoneally, rectally, intravenously, intramuscularly, subcutaneously, Intracerebroventricular injection, and the like.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of cancer.

Food composition

The present invention provides a food composition for preventing or ameliorating a cancer containing a pine tree leaf extract.

The pine cone leaf extract used in the food composition of the present invention can be used as it is as described in the pharmaceutical composition.

In addition, the cancer may be a digestive organ described in the pharmaceutical composition.

The food composition of the present invention can be used as a health functional food. "Health functional food" refers to foods prepared and processed using raw materials or ingredients having useful functions in the human body pursuant to Law No. 6727 on Health Functional Foods, and the term "functional" It is intended to take nutrients for the purpose of controlling nutrients and obtaining effects that are useful for health use such as physiological action.

The food composition of the present invention may contain conventional food additives and, unless otherwise specified, the suitability of the food additives as "food additives" is to be determined by the Food and Drug Administration in accordance with the general provisions of the Food Additives Ordinance approved by the Food and Drug Administration, Of the present invention.

Examples of the substances described in the above-mentioned "Food Additive Code" include natural compounds such as ketones, chemical compounds such as glycine, potassium citrate, nicotinic acid and cinnamic acid, persimmon extracts, licorice extract, crystalline cellulose, high- - Mixed preparations such as a sodium glutamate preparation, a noodle-added alkaline agent, a preservative preparation, a tar coloring agent and the like.

The food composition of the present invention can be used for various foods and beverages to prevent or ameliorate cancer and can be used for various foods, beverages, gums, tea, vitamin complex, health functional supplement, food additive, Powders, granules, tablets, capsules or beverages. May be in any food form other than those described above.

The food composition of the present invention may contain about 1 to 80% by weight, preferably 20 to 60% by weight, of the pine needle leaf extract based on the total weight of the composition for the purpose of preventing or improving cancer. This range may vary depending on the individual's weight, age, sex, health status, diet, and cancer severity.

In addition, the food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and rings for the purpose of preventing or improving cancer.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture of pine needle leaf extract, excipient, binder, disintegrant, and other additives by a conventional method and then compression- Can be directly compression molded. The health functional food of the tablet form may contain a mating agent and the like if necessary, and may be sieved to a suitable skin care agent if necessary.

The hard capsule of the capsule type health functional food may be prepared by filling a normal hard capsule with a mixture of pine needle leaf extract and additives such as excipients or the granular material or the granulated material. And additives such as excipients and the like may be filled in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of pine needle leaf extract, excipient, binder, disintegrant and the like in an appropriate manner. If necessary, the skin may be mixed with white sugar or other suitable skin care agent or a starch, talc or suitable substance It is possible that the

The granular health functional food may be prepared by granulating a mixture of pine needle leaf extract, excipient, binder, disintegrant and the like by a suitable method and, if necessary, a flavoring agent, a mating agent and the like. In the granular form of health functional foods, when the next granularity test was carried out using the No. 12 (1680 μm), No. 14 (1410 μm) and No. 45 (350 μm) sieve, the total amount of the 12- 5.0% or less and that passing through the 45-well body may be 15.0% or less of the total amount.

The definitions of the above excipients, binders, disintegrants, lubricants, mating agents, flavoring agents, and the like are described in documents known in the art and include the same or similar functions (Korean Pharmacopoeia, College of Pharmacy, 5th ed., P. 33-48, 1989).

As described above, the pine leaf extract of the present invention inhibits the growth of cancer cells and exhibits an anti-cancer effect. Therefore, it can be usefully used for the production of a health functional food or a medicine having excellent anticancer effect.

Fig. 1 shows the cell growth of pine needle leaf extract treated with various concentrations in colorectal cancer cells.
FIG. 2 shows the results of Crystal violet staining for the proliferation of pine needle leaf extract treated with HCT116, a colorectal cancer cell line, for 5 days.
Fig. 3 shows the results of treatment with P. vivax leaf extract for 5 days in HCT116 colon cancer cell line. The cell proliferation was examined by crystal violet staining followed by dissolution in 70% ethanol.
FIG. 4 is a graph showing cell growth by BrdU Assay when treated with P. intestinalis leaf extract at various concentrations for 24 hours and 48 hours in HCT116, a colon cancer cell line.
FIG. 5 is a graph showing the cell mobility when treated with shoemole leaf extract according to the concentration in HCT116, a colorectal cancer cell line, through a Wound Healing Assay used as a model of metastasis.

Hereinafter, the present invention will be described in more detail through examples and test examples. However, these examples and test examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples and test examples.

[ Manufacturing example ] Preparation of pine tree leaf extract

Fresh pine tree leaves (Gapyeong-gun, Gangwon-do, Gyeonggi-do, Hongcheon-gun) were collected and the disease, insect eaten and dried were selectively removed and the selected pine tree leaves were washed three times with flowing water in a washing tank. The washed shrub leaves were shaded (shade) for 3 days and cut to a constant size of about 10 mm using a cutting machine. The cut pine trees were transferred to a distillation tank. In the distillation tank, the ratio of the pine tree leaf and the purified water was set to about 1: 2, and the extraction temperature of the pine cones of the distillation tank was maintained at 90 ° C for 3 to 4 hours. At this time, the temperature of the cooling water supplied to the inside of the cooler was maintained at about 18 ° C, and the condensate passing through the cooler was collected in the condensate separator. The collected condensates were mixed with oil and water, so they were subjected to further concentration and filtration to obtain high purity extracts. The obtained extract was a clear, colorless transparent oil and showed a pine wood aroma. The final extract obtained by the above extraction process was aged for 30 days in a cold dark place for balance (stabilization) and flavor and aroma of the final chemical components.

[ Experimental Example ] Anticancer Effectiveness of Pine Needle Leaf Extract in Colon Cancer Cell Lines

The colon cancer cell lines HCT116 and SW620 were dispensed into 96-well plates at a density of 1 × 10 4 cells per well and cultured for 24 hours. The growth inhibition effect on the cancer cell line was confirmed by MTT cytotoxicity assay after culturing the pine needle leaf extract (EOPK) at various concentrations for 24 hours (FIG. 1).

As shown in Fig. 1, it was confirmed that when the pine needle leaf extract had a certain concentration or more, it inhibited cancer cell growth.

1, the following experiment was carried out by setting the concentration of the pine cone leaf extract (EOPK) not toxic to the colon cancer cell line HCT116.

HCT116, a colorectal cancer cell line, was dispensed into a 6-well plate at a density of 5 × 10 4 cells per well and cultured for 24 hours. Pine needle leaf extract (EOPK) was treated at different concentrations and cultured for 24 hours. After 5 days of incubation, the cells were fixed with 1% glutaraldehyde solution and stained with 0.05% Crystal violet solution (Fig. 2).

The absorbance was also measured by dissolving in 70% ethanol and using a microplate reader (Molecular Devices Co.) at 570 nm to 405 nm as a reference (FIG. 3).

As shown in FIG. 2 and FIG. 3, the extract of P. intestinalis inhibited the proliferation of colon cancer cells in a concentration-dependent manner.

HCT116, a colorectal cancer cell line, was divided into 1 × 10 4 cells and 5 × 10 ³ cells in 96-well plates, cultured for 24 hours, and treated with P. intestinalis leaf extract (EOPK) for 24 hours and 48 hours The growth of colon cancer cell lines was confirmed using a BrdU ELISA Kit (kit for measuring the degree of cancer cell proliferation) (FIG. 4).

As shown in FIG. 4, the extract of Pine Pinus species inhibited the proliferation of colon cancer cells in a concentration-dependent manner, and the pine leaf extract was more effective than the pine leaf extract for 24 hours.

In addition, HCT116, a colorectal cancer cell line, was divided into 4 × 10 ⁵ cells in a 6-well plate and cultured for 24 hours. The cell mobility was determined by the Wound Healing Assay (Fig. 5).

As shown in FIG. 5, it was confirmed that the extract of P. vivax suppresses the mobility of colon cancer cells in a concentration-dependent manner.

As can be seen from the above experiment, the extract of Pine Pinus koraiensis shows anti-cancer effect through inhibition of cancer cell growth in HCT116, a cancer cell model in vitro.

Claims (7)

A pharmaceutical composition for preventing or treating cancer containing a pine leaf extract. The pharmaceutical composition according to claim 1, wherein the Pine leaf blade extract is obtained by steam distillation extraction. 3. The pharmaceutical composition according to claim 2, wherein the Pine leaf blade extract is a colorless transparent oil. The pharmaceutical composition according to claim 1, wherein the cancer is gastrointestinal cancer. The pharmaceutical composition according to claim 4, wherein the gastrointestinal cancer is selected from the group consisting of esophageal cancer, stomach cancer, pancreatic cancer, liver cancer, gallbladder cancer, colon cancer and anal cancer. 5. The pharmaceutical composition according to claim 4, wherein the gastrointestinal cancer is colon cancer. A food composition for prevention or improvement of cancer containing Pine leaf extract.

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