KR20200059921A - Composition for preventing or treating cancer comprising fruit extracts of alnus sibirica or fraction thereof - Google Patents

Abstract

According to a composition for preventing or treating cancer containing an Alnus japonica fruit extract or a fraction thereof as an active component, it is possible to provide the composition for preventing or treating cancer which has excellent effects of preventing or treating cancer by inhibiting the expression of MMPs, and at the same time has no side effects when taking the composition, and also contains an economically burdenless natural substance as a main component.

Description

Composition for prevention or treatment of cancer comprising alderberry extract or a fraction thereof as an active ingredient {COMPOSITION FOR PREVENTING OR TREATING CANCER COMPRISING FRUIT EXTRACTS OF ALNUS SIBIRICA OR FRACTION THEREOF}

The present invention relates to a composition for the prevention or treatment of cancer comprising the Alderberry extract or a fraction thereof as an active ingredient, and more specifically, the duck having an excellent effect of inhibiting cancer cell death and cancer metastasis using an extract of Alderberry fruit. It relates to a composition for the prevention or treatment of cancer comprising a tree fruit extract or a fraction thereof as an active ingredient.

As aging progresses, humans show visual characteristics such as gray hair, blotch, wrinkles, and atherosclerosis, Alzheimer's diabetes, cancer, and imbalance of the immune system, resulting in death.

Cell aging, which is one of the causes of human aging, is transformed into cancer cells by irregularly modifying the intracellular division cycle, causing imbalance in the body.

Cancer cells, which repeatedly divide and grow due to mutations in genes that control the cell cycle, use excessive nutrients around them, and if nutrients are deficient afterwards, they metastasize or infiltrate into surrounding tissues and act as malignant tumors. Effect.

MMPs, which are proteolytic enzymes, play an important role in the function of penetrating and spreading the surrounding tissues of cancer.

MMPs are important enzymes involved in regeneration, embryogenesis, angiogenesis, or tumor cell suicide suppression, growth factor activation proteins, and are involved in the breakdown of extracellular matrix and cell migration in vivo.

Cancer cells use MMPs to induce the breakdown of the extracellular matrix, a structural support tissue, and cause the basement membrane to collapse, allowing cells to spread to other tissues. Among the MMPs proteins that cause invasive growth and metastasis of cancer cells, MMP-2 and MMP-9 act on enzymes that break down gelatin, a component that forms the basement membrane and the extracellular matrix.

Inhibitors with basic frameworks of hydroxamate and collagen, zinc binding functional groups developed to inhibit MMPs, which play an important role in the metastasis of cancer cells, act on MMP1, MMP2, MMP7 and MMP9.

They can inhibit MMPs by binding to the zinc ion active site, and non-hydroxamate MMP inhibitors are still in the spotlight as treatments for some diseases to compensate for their shortcomings.

Besides. As the most recent cancer metastasis inhibitor, it helps to move in the case of normal cells, but in order to cause metastasis in cancer cells, filopodia targets the excessive secretion of integrin protein, and the surface is coated with a peptide as a gold nano Treatments that suppress metastasis by using a method that wraps the material and irradiates it are emerging.

Currently, more than 50 MMPs inhibitors have been developed, but there is insufficient development of effective therapeutic agents capable of substantially inhibiting MMPs.

(Patent Document 1) KR 10-2018-0116904 A1

An object of the present invention is to provide a composition for the prevention or treatment of cancer, comprising the alderberry extract or a fraction thereof as an active ingredient.

Another object of the present invention is to provide a composition for the prevention or treatment of cancer comprising alderberry fruit extract and tomatidine as active ingredients.

Another object of the present invention is to prevent or treat cancer that contains natural substances that do not cause side effects when taken and have economical burden as a main ingredient while suppressing the expression of MMPs, and thus have excellent prevention or treatment effects of cancer. It is to provide a composition.

In order to achieve the above object, the composition for preventing or treating cancer according to an embodiment of the present invention includes an Alder sibirica fruit extract or a fraction thereof as an active ingredient.

The composition further comprises Tomatidine, which is isolated from the leaves and stems of tomato plants.

The alderberry extract is extracted using an extraction solvent selected from the group consisting of purified water, alcohol having 1 to 10 carbon atoms, and mixtures thereof.

The Alderberry extract is extracted using alcohol as an extraction solvent.

The Alnus sibirica fruit extract or its fraction has excellent cancer cell killing and inhibitory effects on MMP-2 and MMP-9.

The Alnus sibirica fruit extract or its fraction has an excellent inhibitory effect on the expression of p-ERK and p-p38 proteins.

The cancer may be selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, lung cancer, breast cancer, liver cancer, prostate cancer, pancreatic cancer, gallbladder cancer, and biliary cancer.

Food composition for preventing or treating cancer according to another embodiment of the present invention includes a composition for preventing or treating cancer.

The pharmaceutical composition for preventing or treating cancer according to another embodiment of the present invention includes the composition for preventing or treating cancer.

Hereinafter, the present invention will be described in more detail.

The composition for the prevention or treatment of cancer according to an embodiment of the present invention includes an Alder sibirica fruit extract or a fraction thereof as an active ingredient.

More specifically, the fruit of the alder tree is harvested, and the extract and its fraction are separated using the fruit of the alder tree, and this is used as a composition for preventing or treating cancer.

As for the treatment of cancer, chemotherapy is often performed using the anticancer agent as described above, but it is often seen that the anticancer agent is ineffective (resistant). As a basis for the immunization, for example, it may be due to a biological reaction of a cancer patient, such as inactivation of an anticancer agent in the liver, or an improvement in metabolism, or a biochemical change in cancer cell level. In the latter, changes in the membrane transport mechanism of anticancer agents involved in multidrug resistance, amplification of target enzymes or proteins, drug activation mechanism, decrease in enzyme activity, for example, DNA repair mechanism enhancement, anticancer agent inactivation mechanism enhancement It is believed that the back is rising. In addition, there are cases in which many patients have strong side effects and the patient dies due to side effects.

For this reason, there is a high demand for cancer treatment methods with few side effects. Immunotherapy is receiving attention as a treatment with less side effects, but it is difficult to eradicate the tumor with low tumor control. This is due to the immunotherapy using lymphocytes or NK cells, the main chain of immunity, and also because lymphocytes and NK cells are inherently infectious-response cells, the inhibitory effect on solid cancer is small.

Therefore, there is a strong social request for the development of new anti-cancer drugs that are highly effective against solid cancer, have no side effects such as chemotherapy, and are difficult to cause resistance.

As described above, in the case of a chemotherapeutic agent, the likelihood of side effects is very high, and since it causes resistance, the cancer patient often complains of pain due to side effects other than cancer as the treatment period increases.

Thus, according to the present invention, as the natural extract using alder berries is used as a composition for anticancer, unlike chemotherapy, it lowers the occurrence of side effects by taking and does not cause resistance, but has an excellent effect as an anticancer agent. Compositions can be provided.

The alnus sibirica is geographically inhabited in Korea, Japan, China, and Russia. The bark of alder is a traditional Korean medicine in the private sector, and is a tea that detoxifies antipyretics, expectorants, anti-inflammatory drugs, cough medicines, anticonvulsants, and alcohol. Was used. Alder is known to contain diarylheptanoids, tannins, flavonoids and triterpenoids as its main components.

In the present invention, to extract the extract and its fractions using the fruit, not the bark of the alder tree, and to use it as an anticancer agent.

The alderberry extract is extracted by using an extraction solvent selected from the group consisting of purified water, alcohol having 1 to 10 carbons, and mixtures thereof, and more specifically, extraction using alcohol as an extraction solvent.

The alcohol (Ethyl alcohol) is contained in various "alcohol beverages" is also called alcohol (酒 주), or is also called ethyl alcohol.

The extraction solvent may be used 2 to 50 times based on the weight of the sample, preferably 2 to 20 times. For extraction, the sample can be prevented for 1 to 72 hours for leaching in the extraction solvent, and in particular can be left for 1 to 24 hours.

The filtered extract may be a result obtained by concentrating under reduced pressure with a vacuum rotary concentrator, but is not limited thereto, as long as it is an alderberry extract capable of exhibiting the anti-cancer effect of the present invention, and the extract, a diluent or concentrate of the extract, and dried extract All of the dried product obtained by this, or its modifier or refined product are included.

In addition, the anti-cancer composition of the present invention is excellent in preventing or treating cancer even when only the extract of alder tree fruit and its fraction itself is used, but to show a higher cancer preventing or treating effect, tomatidine ( Tomatidine).

The tomatidine is a substance that is separated from the leaves and stems of tomato plants.

Specifically, tomatidine is a glycoalkaloid, which is mainly contained in the leaves and stems of tomato plants, which are annual plants of the dicotyledon family. It is known that tomatidine stimulates cell growth and mTOR signaling and has a recovery effect of skeletal muscles degraded by aging.

The present invention is to use a composite extract obtained by mixing the extract of alder tree fruit and tomatidine, as compared to the case of using alone, according to the mixing action between the components, The anti-cancer effect will increase.

Specifically, by using an extract of alder tree fruit and a complex extract mixed with tomatidine, it suppresses the expression of MMPs, a proteolytic enzyme important for cancer cell invasion and metastasis, and can exhibit excellent cancer prevention or treatment effects. have.

The composite composition of a mixture of tomatidine and alder tree extracts derived from natural products may stop cancer growth by preventing the proliferation and metastasis of cancer cells through the killing effect of cancer cells in vivo.

Until recently, cancer, which shows the highest mortality rate among human diseases, has been actively researched and developed in various cancer diagnosis and treatment methods according to the fatal risk.

In general, cancer is treated by surgical treatment or chemotherapy, but these treatments remove the lymph nodes surrounding the cancer-causing region and the cancer-causing region, and the treatment proceeds. Cause loss.

The importance of cancer cell invasion and metastasis suppression studies to reduce the loss of bio-function is widely known. These infiltration phenomena form in normal cells and move around the body and are activated when they pass through the cell basement membrane and extracellular matrix to monitor cancer or abnormal cells, but cancer cells amplify these activations more than necessary to cause problems.

In order to infiltrate and metastasize cancer cells to other tissues, the basement membrane and extracellular matrix decomposition must be accompanied, so matrix metalloproteinases (MMPs), which are related proteases, work.

In particular, among them, MMP-2 and MMP-9 are gelatins that degrade gelatin among extracellular matrix proteins, collagen 1, 2, 3, fibronectin, laminin, and gelatin. It is known as the most important component for cancer metastasis and infiltration.

The present invention confirmed the efficacy of MMP-2 and MMP-9 secreted from HT1080 cells and their efficacy for protein expression using tomatidine and alderberry extracts.

As a result, the composite composition of the present invention inhibited gelatin decomposition of MMP-2 and MMP-9 and showed a decrease in expression of MMP-2 and MMP-9 according to a decrease in p-ERK at the protein level.

It has also been shown that the composite composition at the cytoplasmic site significantly reduces MMP-2 protein expression.

This is a signaling protein that is important for all physiological phenomena of cells due to external factors such as reactive oxygen species. MAPK (mitogen-activated protein kinase) includes extracellular signal-regulated kinases (ERKs), p38, NH2-terminal kinase (JNK). It is known that when phosphorylation of proteins by MAPK occurs, the expressions of MMP-2 and MMP-9 are regulated by acting on transcription factors NF-kB and AP-1.

Thus, the composite composition of the present invention regulates the expression of MMP-2 and MMP-9 through the p-p38 and ERK pathways.

When cancer metastasizes to other tissues or blood vessels, infiltration occurs by producing new blood vessels. To investigate the degree of infiltration, stimulation with vascular epidermal factor (VEGF) confirmed cell invasion, and as a result, it was confirmed to inhibit cell invasion.

Finally, through the above results, the composite composition of the present invention may exhibit an effect on inhibiting metastasis of cancer cells by reducing the enzyme activity and protein expression of MMP-2 and MMP-9 expressed during cancer metastasis.

Preferably, the composition for the prevention or treatment of cancer may further include Gavinja extract and Blackcurrant extract.

The Gaebija (Cephalotaxus koreana Nakai) is an evergreen tree growing in a valley or in a forest, reaching a height of 3 m. The leaves are linear, arranged in two rows, the ends are sharply sharp, the front is green, and the vein is clear. Flowers bloom in April with Igahwa, and male flowers are flat, and two female flowers run in one place. Fruits are round, ripened in red in August-September of next year, seeds are long oval, brown.

The Magpie (Carpinus cordata Blume) is a deciduous tree growing in the valley, reaching 15m in height, and its twigs are hairy, but gradually disappear. The leaves are ovate or elliptical, pointed to the bottom of the heart, and have sharp teeth at the edges. The flowers bloom in May with a single flower, and the male inflorescence hangs on the tip of the twig with the leaves, and the female inflorescence hangs from the end of the branch. Fruits are oval, oval, ripen in October.

Preferably, the composition for preventing or treating cancer of the present invention includes Alnus sibirica fruit extract, Gavinus extract, blackcurrant extract and tomatidine.

Even if each natural extract is used separately, it is excellent in preventing or treating cancer, but when the natural extract is used in combination, the anti-cancer effect is increased due to the mixing action between the components.

The cancer may be selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, lung cancer, breast cancer, liver cancer, prostate cancer, pancreatic cancer, gallbladder cancer, and biliary cancer, but inhibits cancer cell death, activity of MMP-2 and MMP-9, and p- ERK and p-p38 protein expression can be used for all cancers that may have anti-cancer effects by suppressing the expression, but is not limited to the above examples.

Food composition for preventing or treating cancer according to another embodiment of the present invention may include a composition for preventing or treating the cancer.

As a specific example, a processed food capable of enhancing the anti-inflammatory effect may be prepared using the food composition. Such processed foods include, for example, confectionery, beverages, alcoholic beverages, fermented foods, canned foods, milk processed foods, meat processed foods, noodles, and the like. Cookies include biscuits, pies, cakes, breads, candy, jellies, chewing gum, and cereals. Drinks include drinking water, carbonated beverages, functional ionic beverages, functional ionic beverages, juices (e.g., apples, pears, grapes, aloe, citrus fruits, peaches, carrots, tomato juices, etc.), and sikhye. Alcoholic beverages include sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and red pepper paste. Canned foods include canned seafood (eg, canned tuna, mackerel, saury, turban, etc.), canned livestock products (canned beef, pork, chicken, turkey, etc.), and canned agricultural products (corn, peach, canned file apple, etc.). Milk processed foods include cheese, butter, and yogurt. Processed meat products include pork cutlet, beef cutlet, chicken cutlet and sausage. It includes sweet and sour pork, nuggets, and bread, and so on. Contains noodles such as sealed packaging noodles. In addition to this, the composition may be used for retort foods, soups, and the like.

When using the Alnus sibirica fruit extract, Gavinja extract, blackcurrant extract and tomatidine as functional food composition for anti-cancer, the unique aroma of Alder tree extract, Gavinja extract and blackcurrant extract and The problem of poor palatability occurs due to taste.

In order to prevent such a problem, a functional food composition may be prepared using a chemical component such as a sweetener, but considering that the functional food composition is used for anticancer, natural extracts other than chemical components such as sweeteners are used. To improve palatability.

Thus, preferably, the present invention further includes a licorice extract and a ginseng extract, to enhance palatability and enhance anticancer effects.

The licorice (Glycyrrhiza uralensis Fisch.) Is a perennial plant grown for medicinal purposes, reaching a height of 1 m, with ridges and preoccupations. Leaves are elongated, odd upper right leaf lobe, 7 to 17 lobules each, ovate original, with white hairs on both sides, and preoccupation. The flowers are light purple, and run as a gunshot wound on the leaf axil in July to August. Fruits are narrow, linear, with 6 to 8 seeds.

The terminus (Senna tora (L.) Roxb.) Is an annual plant that grows and reaches a height of 2 m. The leaf is an even numbered dominant biplane with 2 to 4 pairs of lobules, and the lobules are obovate, slightly pointed at the tip, and sharp or original. The flowers are yellow with peduncles, and hang on 1-2 leaf axils in June to August. The fruits are curved like a bow.

Preferably, the composition for anti-cancer according to an embodiment of the present invention includes alder sibirica fruit extract, Gaeja birch extract, blackcurrant extract and tomatidine,

Licorice extract and Cassia tora extract may be further included.

Compared to the case of using the alder tree fruit extract, Gaeja tree extract, blackcurrant extract and tomatidine, when the licorice extract and Cassia tora extract are used in combination, the palatability is improved and the convenience of taking is increased. , When used as an anti-cancer agent, the anti-cancer effect also rises due to the mixing action according to the combined use of each component.

More preferably, with respect to 100 parts by weight of the Alnus sibirica fruit extract, 50 to 100 parts by weight of Gavinja extract, 50 to 100 parts by weight of Magpie extract, 80 to 120 parts by weight of tomatidine, 20 licorice extract It may include 40 to 40 parts by weight and 20 to 40 parts by weight of the extract of the ginseng. In the case of the above range, a synergistic effect of a critical significance level is expressed as a synergistic effect due to the interaction of each extract, and when it is outside the above range, the synergistic effect is rapidly reduced or almost absent. That is, when used as a food composition within the above range, the palatability is increased, and the anti-inflammatory effect is increased by the interaction of each extract.

The pharmaceutical composition according to another embodiment of the present invention includes the anti-cancer composition.

The anti-cancer pharmaceutical composition may further include one selected from the group consisting of pharmaceutically acceptable carriers, excipients, and diluents. Specifically, the pharmaceutical composition is formulated in the form of oral dosage forms, external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. Can be. In the present invention, as a carrier, excipient, and diluent that may be included in the pharmaceutical composition, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate , Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient, such as starch, calcium carbonate, in the extract and its fractions. It is prepared by mixing sucrose or lactose and gelatin. Also, lubricants such as magnesium stearate and talc are used in addition to simple excipients. Liquid preparations for oral administration may include various excipients, such as wetting agents, sweeteners, flavoring agents, preservatives, etc., in addition to water and liquid paraffin, which are simple diluents commonly used for suspending agents, liquid solutions, emulsions, syrups, etc. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, the term "pharmaceutically effective amount" of the present invention to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention Means an amount sufficient, and the effective dose level is the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration, route of administration and rate of excretion of the composition of the invention used. The duration of treatment can be determined by factors including drugs used in combination or coincidental with the composition of the present invention used and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a known immunotherapeutic agent. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects.

According to the composition for preventing or treating cancer comprising the Alderberry extract of the present invention or a fraction thereof as an active ingredient, the expression of MMPs is suppressed, and the prevention or treatment effect of cancer is excellent, and at the same time, no side effects occur when taking , It can provide a composition for the prevention or treatment of cancer, which contains economically burdenless natural substances as a main component.

1 is an experimental result for the cancer cell killing effect of the composition according to an embodiment of the present invention.
2 is an experimental result for inhibiting MMP-9 activity of the composition according to an embodiment of the present invention.
3 is an experimental result for confirming the inhibitory effect of the protein related to the expression of MMPs of the composition according to an embodiment of the present invention.
4 is an experimental result for suppressing cell invasion occurring during cancer metastasis of the composition according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art to which the present invention pertains can easily practice. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein.

[Production Example 1: Preparation of a complex mixture of alderberry extract and tomatidine]

Dulbecco's Modified Eagle's Medium (DMEM), Trypsin-EDTA, penicillin (10,000 U / ml) / streptomycin (10,000 μg / ml) / amphotericin (2,500 μg / ml), fetal bovine serum (FBS) reagent for cell culture is Gibco BRL , Life Technologies (Paisley, Scotland). The HT1080 cell line was purchased from ATCC (American Type Culture Collection, USA). Tomatidine hydrochloride, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) reagent, gelatin and other reagents are available from Sigma Chemical Co. (St. Louis, MO, USA) and santa cruz biotechnology.

Alder fruit extract used in the present invention was extracted by the following method. First, the black rice was washed thoroughly with running water and then dried completely. The washed and dried black rice was pulverized with a blender, produced as a powder, and then extracted for 3 days and then filtered. Each filtered filtrate was concentrated under reduced pressure to obtain an extract of alderberry fruit in powder form. The prepared powder-type sample was mixed with tomatidine and named as TASE, and diluted with dimethyl sulfoxide (DMSO) to a test concentration.

[Experimental Example 1: Review of anti-cancer effects of TASE]

Experiment method

MTT assay

The cytotoxicity of TASE against HT1080 cells was measured using MTT (3- (4,5-dimethyl-2-yl) -2,5-diphenyltetrazolium bromide).

Gelatin zymography

For MMP-2 and MMP-9 activity, HT1080 cells cultured in DMEM medium without FBS were treated with TASE, and experiments were performed with 1 ng / ml of PMA, which is known to stimulate the expression and activity of MMP. The cell culture solution containing 50 μg total protein was electrophoresed using 10% polyacrylamide gels under non-reducing conditions containing 1.5 mg / ml gelatin. Gelatin-decomposed bands appear as transparent bands on a blue background. The intensity of the bands appears in proportion to the activity of the MMPs, and was observed and observed with a LAS3000® image analyzer (Fujifilm Life Science, Tokyo, Japan).

Western blot experiment

Elution buffer (50 mM Tris-HCl, pH 7.5, 0.4% Nonidet P-40, 120 mM NaCl, 1.5 mM MgCl2, 2 mM phenylmethylsulfonyl fluoride, 80 μg / ml leupeptin, in HT1080 cells pre-treated with TASE and stimulated with PMA) 3 mM NaF and 1 mM DTT) was added and treated at 4 ° C. for 30 min. After electrophoresis of 10 μg of cell eluate on 10% Tris-HCl gel, the protein was transferred to a nitrocellulose membrane. Then, 10% skim milk was pre-treated on nitrocellulose membrane, and primary antibodies against the target protein (anti-MMP-9, anti-MMP-2, p-p38, p-JNK, p-ERK) were treated, and 2 After treatment of the secondary antibody, the target protein was detected using a chemiluminescent ECL kit (Amersham Pharmacia Biotech). The band of the Western blot was observed using a LAS3000® image analyzer (Fujifilm Life Science, Tokyo, Japan).

Cell infiltration experiment

Cell infiltration experiment was performed according to the method of Invasion Assay Kit (ECM550) Chemicon®. A cell culture insert was placed in a 24 well cell culture plate, and DMEM medium without FBS was added to the insert, followed by incubation at 36 ° C for 30 minutes. After removing the medium in the insert, DMEM medium to which 10% FBS was added outside the insert was added, and DMEM medium and cells without FBS were added to the insert, followed by incubation at 36 ° C for 1 hour. The cultured cells were pre-treated with TASE, treated with VEGF, and incubated at 36 ° C. for 24 hours. After removing the medium in the 24 well culture plate and the medium in the insert, the remaining cells in the insert were removed using a cotton swab. Cells that penetrated the bottom of the insert were stained, dissolved in 10% acetic acid solution, transferred to a 96-well plate, and absorbance was measured at 560 nm.

Statistics processing

Each experiment obtained the result through three or more repetitive experiments, and was expressed as the mean ± standard deviation for each sample concentration. The significance difference test for each sample concentration group was considered as a statistically significant result after Student's test compared to the control group.

TASE's HT1080 cancer cell killing effect

To investigate the cancer cell killing effect of HT1080 of TASE mixed with Tomatidine and AFEE, an MTT assay was performed. As shown in FIG. 1, the TASE treatment increased the killing effect of cancer cells as the concentration increased compared to the blank test group, and 50% of the cancer cells were killed at 8 μM.

Inhibitory effect of TASE on MMP-9 activity in HT1080 cells stimulated with PMA, a cancer-promoting factor

TASE, a mixture of tomatidine and AFEE, for enzyme activity of MMP-2 and MMP-9, which are essential enzymes for invasion and angiogenesis during cancer metastasis of cancer cells through gelatin breakdown of the extracellular matrix, Gelatin zymography was performed to investigate the effect.

After processing TASE on HT1080 cells, the enzyme activity of MMP-2 and MMP-9 was measured in the supernatant cultured for 72 hours. As shown in FIG. 2, TSSEE was found to decrease the activity of MMP-9 in a concentration-dependent manner at 1 μM or more compared to the PMA-treated group. In addition, it was found that the activity of MMP-2, which is the active form, was reduced at a concentration of 4 μM or more, and the activity was decreased by 52% compared to the PMA treatment group at a maximum concentration of 8 μM.

Inhibitory effect of TASE on proteins related to expression of MMPs in HT1080 cells

The ability of TASE to regulate the protein expression of MMPs, an extracellular matrix degrading protein that causes invasive growth and metastasis of cancer cells, was investigated. As shown in FIG. 3, it was shown that the protein expression of p-p38 was suppressed at a concentration of 8 μM and p-ERK involved in MMP expression when compared to the PMA treatment group at a concentration of TASE of 1 μM or more.

In addition, it was observed that the protein expression of MMP-2 and MMP-9 was also lowered by TASE. TASE was shown to suppress the expression of MMP-2 and MMP-9 by lowering the expression of Erk and p-p38.

Inhibitory effect of TASE on cell invasion during cancer metastasis

A cell invasion assay was performed to investigate the inhibitory effect of TASE on cell invasion occurring during cancer metastasis. The control group and the TASE-treated group were cultured for 24 hours by treatment with vascular epidermal growth factor (VEGF). As shown in FIG. 4, the TASE-treated group was found to inhibit cell infiltration in a concentration-dependent manner compared to the control group VEGF-treated group. TASE was found to inhibit about 51% of cell infiltration compared to the VEGF-treated group at a concentration of 1 μM.

[Production Example 2: Preparation of a complex extract]

Using the same method as the alderberry fruit extract of Preparation Example 1, a gaenja tree extract, a blackcurrant extract, a licorice extract and a ginseng extract were prepared.

Alder tree extract (TA), Gaeja tree extract (RE), blackcurrant extract (TU), tomatidine (SE), licorice extract (CU) and Cassia tora extract (CA) are mixed as shown in Table 1 below. Was prepared.

TASE TASE2 TASE3 TASE4 TASE5 TASE6 TASE7 TASE8 TASE9 TA 100 100 100 100 100 100 100 100 100 RE - 40 50 70 100 120 70 70 70 TU - 40 50 70 100 120 70 70 70 SE 100 100 100 100 100 100 100 100 100 CU - - - - - - 10 30 50 CA - - - - - - 10 30 50

(Unit parts by weight) [Experimental Example 2: Review of anticancer effect]

With respect to the contents reviewed in Experimental Example 1, the same experiment was conducted for TASEs 2 to 6 to confirm the results.

The evaluation of TASE was set to an index of 5, and the anticancer effect on TASE 2 to 6 was evaluated by a relative score and expressed as an index. The results are shown in Table 2 below.

TASE TASE2 TASE3 TASE4 TASE5 TASE6 Cancer cell killing effect 5 4 6 7 8 6 Inhibition of MMP-9 activity 5 4 6 6 9 5 Inhibition of p-ERK and -p38 expression 5 3 5 7 8 5 Cell infiltration inhibition 5 4 6 7 9 5

As shown in Table 2, it was confirmed that the anti-cancer effect was equal or higher in TASE3 to 5 compared to TASE.

[Experimental Example 3: Sensory evaluation]

Sensory evaluation was performed for TASE and TASE 4 shown in Table 2 above. In addition, TASE 7 and 8 were also evaluated in the same way.

TASE, TASE4, TASE7, and TASE8 were prepared as teas, and 20 adult males and females were asked to evaluate their flavor, taste, and preference. Evaluation was requested with a score from 1 to 10, and the evaluation result for 20 persons was converted into an average score.

TASE TASE4 TASE7 TASE8 flavor 4 3 7 8 incense 4 5 8 8 Preference 4 3 7 8

As shown in Table 3, in the case of TASE4, which was evaluated as having excellent anti-cancer effect, it was confirmed that the preference was lowered due to the natural flavor and taste of the natural extract itself. On the other hand, it was confirmed that TASE 7 and 8 exhibited excellent evaluation in taste, aroma, and preference.

This can enhance the palatability by reducing the flavor and taste of the other natural extracts of the licorice and ginseng extract contained in TASE 7 and 8.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also provided. It belongs to the scope of rights.

Claims (9)

Alnus sibirica fruit extract or its fraction containing as an active ingredient
Composition for the prevention or treatment of cancer. According to claim 1,
The composition further comprises Tomatidine,
The tomatidine is to separate from the leaves and stems of tomato plants
Composition for the prevention or treatment of cancer. According to claim 1,
The alderberry extract is extracted using an extraction solvent selected from the group consisting of purified water, alcohols having 1 to 10 carbon atoms, and mixtures thereof.
Composition for the prevention or treatment of cancer. According to claim 1,
The Alderberry extract is extracted using alcohol as an extraction solvent.
Composition for the prevention or treatment of cancer. According to claim 1,
The Alnus sibirica fruit extract or its fraction has excellent cancer cell killing and inhibitory effects on MMP-2 and MMP-9.
Composition for the prevention or treatment of cancer. According to claim 1,
The alnus sibirica fruit extract or a fraction thereof inhibits the expression of p-ERK and p-p38 proteins
Composition for the prevention or treatment of cancer. According to claim 1,
The cancer is selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, lung cancer, breast cancer, liver cancer, prostate cancer, pancreatic cancer, gallbladder cancer, and biliary cancer.
Composition for the prevention or treatment of cancer. Claims 1 to 7 comprising a composition according to any one of claims
Food composition for the prevention or treatment of cancer. Claims 1 to 7 comprising a composition according to any one of claims
Pharmaceutical composition for the prevention or treatment of cancer.

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