KR20130032720A - A composition comprising the extract of melia azedarach showing anti-cancer activity against stomach tumor - Google Patents

Abstract

PURPOSE: A composition containing a Melia azedarach L. extract is provided to ensure remarkable anticancer activity in vivo and in vitro without side effects, and to prevent and treat gastric cancer. CONSTITUTION: A pharmaceutical composition for preventing and treating gastric cancer contains a Melia azedarach L. extract as an active ingredient. The extract is prepared using water including purified water, C1-C4 alcohol, or a mixture thereof. The extract suppresses SNU-16 cells and proliferation of tumor induced from the cells. [Reference numerals] (a) Injection concentration of Melia azedarach L. extract; (b) Cancer volume = (cancer length x cancer width^2)/2;

Description

Composition comprising the extract of Melia azedarach showing anti-cancer activity against stomach tumor}

The present invention relates to a composition for the prevention and treatment of gastric cancer, and to a composition having significant in vitro and in vivo anticancer effects utilizing SNU-16 cells.

The composition for the prevention and treatment of gastric cancer of the present invention has a technical feature of including the extract as an active ingredient.

Cancer is also called a malignant tumor, malignant neoplasm, carcinoma, and sarcoma, leukemia, and lymphoma are also included in the category of cancer. .

In particular, gastric cancer has the highest incidence in Korea, and males account for 30% of all cancers, and females account for 15% after cervical cancer.

Therefore, the incidence of gastric cancer accounted for about 50% of all male and female cancers.

Also, like stomach cancer takes place via a complex, multi-step process that involved a variety of risk factors, risk factors of gastric cancer is high salt diet, fat is burnt food, tobacco, alcohol, and Helicobacter pylori (Helicobacter pylori) infections, genetic factors Known.

When people are exposed to these carcinogens in their daily lives, the carcinogenic process is affected by the nature and degree of exposure of the carcinogen itself.

On the other hand, the present invention relates to a composition having anti-cancer activity against gastric cancer, which contains the extract as an active ingredient, it will be described in relation to the following.

The Melia azedarach L. var. Japonica Makino is a dried, mature fruit of the Meliaceae family. It is large, full, and its bark is golden and its pulp is yellowish white.

Appearance is spherical ~ oval, 10 ~ 15mm in diameter, outer surface is yellowish brown, glossy and wrinkled, transverse surface is light yellow, divided into 4 ~ 5 threads, and there is one seed in it.

Main ingredients include limonoid, tosendanin, alkaloid, azaridine, and other ingredients such as resin, tannin, and benzoic acid. have.

The main pharmacological action is used for the purpose of antiparasitic action and antimicrobial action, and in oriental medicine, it is hard to settle, settles, and enters the liver, which leads to novel fever. It is effective in Haeulji pain, and it can be used for thorax narrowing abdominal pain due to liver gastrointestinal pain, sensation pain, and pentagonal pain. It is known to be used for the disease.

On the other hand, although the conventional patent documents that use the cheonngi for anti-cancer activity against gastric cancer have not been searched. Looking at the patent document utilizing the cheonnyeon, Korean Patent Publication No. 10-2001-0093861 discloses a cosmetic composition for whitening containing the cheonnyun extract This is described.

Second, Korean Laid-Open Patent Publication No. 10-2010-0011453 describes the use of cheonnyeonja extract for oxidative stress and inflammatory diseases, which is induced by DPPS radical, NO scavenging activity, and LPS in RAW 264.7 cells. Experimental confirmation of the inhibitory effects of NO, PGE2, iNOS, COX-2 and inflammatory cytokines is described.

Third, Korean Laid-Open Patent Publication No. 10-2010-0011449 discloses that the effect of the anti-inflammatory experiment was confirmed by using a complex extract of Chunjaon and Hyunho color.

Fourth, Korean Laid-Open Patent Publication No. 10-2011-0080184 describes a composition for preventing and treating a Helicobacter bacterial infection using a Stylus extract.

In summary, although it has been found that some of the medicinal herb improve the oxidative stress, inflammation, and Helicobacter disease, the anticancer activity against gastric cancer has not been studied sufficiently.

In this regard, the present applicant has developed a medicinal herb extract agent that exhibits excellent anticancer effect against gastric cancer, and the anticancer effect against gastric cancer in vitro and in vivo experiments was confirmed. To complete.

In particular, the applicant used SNU-16 cells sold by the Korea Cell Line Bank, SNU-16 cells are floating cells growing in the medium as human gastric adenocarcinoma cells.

On the other hand, a paper using SNU-16 cells, “Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja ( Citrus grandis ) Leaves” (Jeong Yong Moon et al., J. Korean) Soc. Appl. Biol. Chem. 52 (2), 168-175, 2009.) describes the application to SNU-16 cells utilizing extracts of Angelica leaf.

KR 10-2001-0093861 A KR 10-2010-0011453 A KR 10-2010-0011449 A KR 10-2011-0080184 A

“Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja (Citrus grandis) Leaves” (Jeong Yong Moon et al. 5, J. Korean Soc. Appl. Biol. Chem. 52 (2) , 168-175, 2009.) “Anti-cancer Effects of Water and Ethanol Extracts from Persimmon Leaves on Korean Gastric Cancer Cell Lines” (Kim, Ho-Jung, Mi-Kyung Kim, Korean Journal of Nutrition, Vol. 36, No. 2, p. 133-146, 2003.3) “Study on Anticancer Effects in Nude Mice of Genetic Recombinant Interferon a-2a” (Kim Chang-Hwan et al. 7, Journal of the Korean Society for Laboratory Animal Science, 14, pp. 251-255, 1998)

An object of the present invention is to provide a composition and health functional food containing as an active ingredient cheonnyeonja extract having anticancer activity against stomach cancer.

The present invention, by providing a pharmaceutical composition for the prevention and treatment of gastric cancer containing a cheonnyeonja extract as an active ingredient, to solve the technical problem.

The gastric cancer is characterized in that it occurs in mammals such as pigs, cows, goats, including humans.

The puncture extract of the present invention may be prepared by cutting the dried puncturer into water containing about 1 to 20 times the dry weight, preferably about 2 snacks and 6 times (v / w) purified water, C ₁ to C ₄ lower alcohols, or With these mixed solvents, preferably water, cold extraction, heat extraction, ultrasonic waves for about 1 hour to 10 days, preferably about 2 hours to 5 hours at an extraction temperature of 20 to 150 ℃, preferably 70 to 120 ℃ Extraction using extraction, such as reflux cooling extraction, preferably by reflux cooling extraction and then concentrated under reduced pressure to obtain the extract of the present invention.

The pharmaceutical composition containing the extract of the present invention comprises the extract in an amount of 0.1 to 50% by weight based on the total weight of the composition.

However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

Extract itself of the present invention is a drug that can be used with confidence even for long-term administration for the purpose of prevention because there is little toxicity and side effects.

The composition of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral dosage forms, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents that may be included in the composition comprising extracts include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy, hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid form preparations contain at least one or more excipients, for example, calcium carbonate, sucrose, etc. Or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.

Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsion lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used.

As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compositions of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the dry matter of the present invention is preferably administered in 0.5g / kg to 5g / kg, preferably 1g / kg to 3g / kg per day. Administration can be done once a day or divided several times. Accordingly, the dosage is not limited in any way to the scope of the present invention.

In addition, the composition of the present invention can be administered to various mammals such as rats, mice, livestock, humans. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

The present invention provides a health functional food for the prevention and improvement of gastric cancer caused by gastric cancer cell proliferation, which contains the extract of Chunjaun obtained as an active ingredient.

The composition comprising the extract of the present invention can be used in a variety of drugs, food and beverages for the prevention and improvement of gastric cancer caused by gastric cancer cell proliferation. Foods to which the extract of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Extract of the present invention may be added to food or beverage for the purpose of preventing and improving gastric cancer caused by gastric cancer cell proliferation. At this time, the amount of the extract in the food or beverage is generally the health food composition of the present invention can be added to 1 to 5% by weight of the total food weight and the health beverage composition is 0.02 to 10g, preferably 0.3 based on 100ml To 1 g.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient, as long as it contains the extract as an essential ingredient at the indicated ratio, and there is no particular limitation to the liquid ingredient.

Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, ilgin acid and salts thereof, organic acids , Protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The composition containing the extract of the present invention as an active ingredient has a remarkable in vitro and in vivo anticancer activity effect.

Compositions and health functional foods containing the present inventors cheonnyeonja extract as an active ingredient, by taking continuously without bringing side effects, the effect of preventing and treating gastric cancer is remarkable.

1 is a diagram of in vitro SNU-16 cells.
FIG. 2 is a diagram showing the results of in vivo anticancer effects according to Experimental Example 2. FIG.

The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may properly define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

Therefore, the examples, reference examples, experimental examples, formulation examples, and drawings described in the present specification are merely the most preferred examples of the present invention and do not represent all the technical ideas of the present invention. Therefore, It should be understood that various equivalents and modifications may be substituted for those embodiments.

Example 1.Preparation of Stellar Extract

The trainer used in this example used a dried trainer purchased from University of Korea.

First, using a grinder (Daesung Akron, DA700) to pulverize the particles to a size of 30 mesh or less to obtain a puncture powder, distilled water corresponding to three times (v / w) of the puncture powder (1 kg) An aqueous 70% ethyl alcohol solution was added thereto, followed by reflux extraction at 100 ° C. for 3 hours.

Next, the resultant was filtered and concentrated under reduced pressure, and then lyophilized to obtain 600 g of a powdery Stellar extract, which was used as a test sample.

Reference Example 1. Preparation of in vitro anticancer experiment

1-1. SNU-16 Cell Preparation

SNU-16 cells used in the experiments are the most suitable model for in vitro experiments to investigate the anticancer effect as cells isolated from human gastric cancer.

SUN-16 cells were cultured by the Korea Cell Line Bank and RPMI 1640 (Gibco, USA) medium was used for cell culture, and 10% Fetal Bovine Serum (FBS, Gibco, USA) was used.

1 shows in vitro SNU-16 cells.

1-2. Sample Preparation

The test sample was prepared by dissolving the puncture extract according to Example 1 in Dimethylsulphoxide (DMSO) and diluting with RPMI 1640 medium.

First, a stock solution of the extract of St. Panaxella was prepared at a concentration of 100 mg / ml, and then diluted with RPMI 1640 medium to prepare a final sample for cytotoxicity experiment.

1-3. Composition

After preparing the stock solution at a concentration of 100 mg / ml, the step was diluted with RPMI 1640 medium, and the samples were respectively 0.1 mg / ml, 0.5 mg / ml, 1 mg / ml, 5 mg / ml, and 10 mg / ml. The group was composed of the concentration.

Experimental Example 1. In vitro anticancer effect of Chunja extract

First, Reference Example 1 and Experimental Example 1 are the experimental methods described in the existing document (“Anti-tumor Effect of Persimmon Leaf Water and Ethanol Extracts on Korean Gastric Cancer Cell Lines” (Kim Ho-jeong, Kim Mi-kyung, Korean Journal of Nutrition, 36, No. 2, p. 133-146, 2003.3)).

The SNU-16 cells prepared in Reference Example 1 were prepared at a concentration of 10 ⁶ cells / ml for each well, and 100 μl of each well of each 96well plate was uniformly added thereto. Mg / ml, 0.5 mg / ml, 1 mg / ml, 5 mg / ml, 10 mg / ml) were added in 100 μl, and control was put in 100 μl of RPMI 1640 medium instead of the sample solution.

Subsequently, the 96 well plate was placed in a cell culture incubator (37 ° C., 5% CO ₂ ) and incubated for 72 hours, followed by MTT (3- [4,5-dimethy thizol-2-yl] -2,5-diphenyl tetrazolium After application of bromide, sigma), 10 μl of MTT (5 mg / ml PBS) was added to each well for 4 hours, and then placed in an incubator for cell culture (37 ℃, 5% CO ₂ ) again after 4 hours. Fluorometer (FLUO star Galaxy, Germany) at 540 nm.

Three samples were measured per concentration of each sample and repeated three times per cell.

Table 1 shows the results of the in vitro anticancer effect according to Experimental Example 1.

SNU-16 O.D value Viability Cell 0.844 100.0 0.1 0.208 24.6 0.5 0.168 19.9 One 0.907 107.5 5 0.000 0.0 10 0.000 0.0

Referring to Table 1, it can be seen that the St. Panax extract has anticancer activity against SNU-16 gastric cancer cells, and is generally concentration-dependent.

Reference Example 2 in vivo (in vi vo) preparing anti-cancer Experiment

2-1. SNU-16 Cell Preparation

In the same manner as in Reference Example 1, SNU-16 cells were prepared.

2-2. Sample Preparation

The test sample was used by dissolving the puncture extract according to Example 1 in Dimethylsulphoxide (DMSO) and then diluting according to the group.

2-3. Laboratory Animal Preparation

Twenty five-week-old male BALB / c-nu / nu mice were purchased and used for experiments after 1 week of pure breeding.

2-4. Composition

The experimental animals were divided into three groups of five animals per group, and then grouped into tumor group inoculation group, no treatment group, 1000 mg / kg administration group, and 500 mg / kg administration group.

Experimental Example 2 in vivo (in vi vo) antitumor effects of extracts thousand ryeonja

First, Reference Example 2 and Experimental Example 2 are experimental methods described in the existing literature (“Study on anticancer effects in Nude Mice of Genetic Recombination Interferon a-2a” (Kim Chang-Hwan et al. -255, 1998).

SNU-16 cell suspension (2 × 10 ⁶ cells / ml) prepared in Reference Example 2 was injected subcutaneously in the rat subcutaneous subcutaneously at 200 ul dose and waited until tumors grew to a sufficient size.

The grown tumor tissue was extracted, cut into 2-3 mm in diameter, placed in a trocar for transplantation, and the tumor tissue was implanted subcutaneously under the right shoulder scapula by 1 slice per animal.

In the experiment, after inoculating tumor tissue into the animal, the size of tumor tissue was 100mm³More than adult From the next day, five animals per group were divided into three groups (untreated group after tumor cell inoculation, 100 mg / kg administration group and 500 mg / kg administration group per day) and administered orally to the administration group for 21 days.

Tumor volume were calculated as V = ab ^2/2 a and b was measured three times the size of the tumor at 1 week in order to determine the anticancer effect of each mean the tumor length and width, and cloth ryeonja extract.

Figure 2 is a diagram showing the results of in vivo anti-cancer effect according to Experimental Example 2, showing that the puncture extract has anti-cancer activity against gastric cancer tissues induced in SNU-16 gastric cancer cells, especially in the dose It can be seen that it is excellent in proportion.

In summary, Experimental Example 1 and Experimental Example 2, it can be said that the cheonnyeonja extract is excellent in vitro and in vivo anti-cancer activity, and generally proportional to the concentration and dose.

Preparation Example 1. Preparation of powder

Angelica Extract (CC-1) 20 mg

Lactose 100 mg

10 mg of talc

The above components are mixed and filled in airtight bags to prepare powders.

Formulation Example 2. Preparation of tablets

Millennium Extract (CC-1) 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation Example 3. Preparation of capsules

Millennium Extract (CC-1) 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2mg

The capsules are prepared by mixing the above components and filling the gelatin capsules according to a conventional manufacturing method.

Formulation Example 4. Preparation of injection

Millennium Extract (CC-1) 10 mg

180 mg mannitol

Sterile Distillers 2974 mg for injection

Na ₂ HPO _4, 12H20 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation Example 5 Preparation of Liquid

Angelica Extract (CC-1) 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, and lemon flavor is added thereto, then the above ingredients are mixed, adjusted to 100 ml by adding purified water, and then filled into a brown bottle and sterilized to prepare a liquid solution. do.

Formulation Example 6 Preparation of Healthy Food

Millennium Extract (CC-1) 1000 mg

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg vitamin B1

0.15 mg of vitamin B2

0.5 mg vitamin B6

0.2 [mu] g vitamin B12

10 mg vitamin C

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

Potassium Citrate 90 mg

100 mg of calcium carbonate

24.8 mg of magnesium chloride

The composition ratio of the above-mentioned vitamins and mixed substances is mixed with a component suitable for a health food as a preferable example, but the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method, and then granulated. It can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation Example 7 Preparation of Healthy Drink

Millennium Extract (CC-1) 1000 mg

Citric acid 1000 mg

Oligosaccharide 100 mg

Plum concentrate 2 g

1 g of taurine

Purified water was added to the mixture,

After mixing the above components according to a conventional healthy beverage production method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained by sterilization in a sterilized 2 L container, sealed sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Claims (4)

Pharmaceutical composition for the prevention and treatment of gastric cancer containing Melia azedarach L. var. Japonica Makino as an active ingredient.

The method of claim 1,
The extract is water, including purified water, C ₁ to C ₄ lower alcohol, or a mixture thereof, characterized in that the extract available in the solvent, the pharmaceutical composition for the prevention and treatment of gastric cancer. The method of claim 1,
The extract is characterized in that to inhibit the proliferation of SNU-16 cells and the tumor induced by the cells, pharmaceutical composition for the prevention and treatment of gastric cancer. Health functional food for the prevention and improvement of gastric cancer containing Melia azedarach L. var. Japonica Makino extract as an active ingredient, but adding a food additive that is acceptable to food.

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