JP2003238422A - Apotosis-inducing agent and health food - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new apotosis-inducing agent and health food which include natural food materials. <P>SOLUTION: The apotosis-inducing agent includes at least one selected from propolis extract, grape skins and hawthorn extract, particularly they are useful as an apotosis inducing agent for gastric cancer cells. The agent and the food materials are used in the form of capsules, e.g. hard capsules or soft capsules, tablets, pills, powder, granules or candy. As recognized in S1, S2 and S3, the gastric cancer cells are fragmented by the action of the apotosis-inducing agent and it is confirmed that the apotosis is induced. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an apoptosis inducer containing a natural food material and a health food containing the same. More specifically, it relates to an apoptosis inducer that is particularly effective against gastric cancer cells and a health food containing the same.

[0002]

2. Description of the Related Art Cancer still has the highest mortality rate among lifestyle-related diseases, and there is no specific medicine. Therefore, research on prevention and treatment of cancer is being actively conducted.
Cancer is also called by various names such as gastric cancer, liver cancer, and lung cancer depending on the site of its origin. Gastric cancer accounts for about 30% of all cancers, and particularly prefers salty foods and hot foods. People who eat are said to be more likely to have stomach cancer.

Currently, drugs used for cancer patients include alkylating agents such as nitrogen mustard and cyclophosphamide, purine antimetabolites such as 6-mercaptopurine and azathioprine, and 5-fluorouracil. There are pyrimidine antimetabolites such as and sirabitane, antifolate antimetabolites such as methotrexate and aminopterin, and antitumor antibiotics such as mitomycin C and bleomycin, but they all suppress the nucleic acid synthesis system. It is toxic to normal cells and causes side effects such as hematopoietic disorders.

On the other hand, as foods having an anti-cancer action, health foods containing a large amount of polysaccharides having an immunostimulating action such as Agaricus edulis and Phellinus linteus are commercially available, and people are conscious of prevention and treatment of cancer in the world. Therefore, the development of pharmaceuticals and health foods having high anticancer activity is desired.

[0005]

SUMMARY OF THE INVENTION It is therefore an object of the present invention to provide an apoptosis inducer which is less toxic to normal cells and less likely to cause side effects such as hematopoietic disorders, and a health food containing the same.

[0006]

[Means for Solving the Problems] In view of the above situation, the present inventors have conducted a screening to find a food material containing a substance that induces apoptosis in cancer cells Propolis extract,
The inventors have found that the grape skin extract and the hawthorn extract have the above-mentioned effects, and have completed the present invention. That is, the cancer cell apoptosis-inducing agent according to claim 1 is a cancer cell apoptosis-inducing agent containing at least one food material selected from the group consisting of propolis extract, grape skin extract and hawthorn extract. Claim 2
The cancer cell apoptosis-inducing agent according to claim 1, wherein the cancer cell is a gastric cancer cell. The health food according to claim 3 is claim 1.
Alternatively, it is a health food containing the apoptosis inducer described in 2.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION A cancer cell apoptosis-inducing agent according to an embodiment of the present invention will be described below. It should be noted that the configurations described below do not limit the present invention and can be variously modified within the scope of the gist of the present invention.

The present invention is an apoptosis inducer for cancer cells containing at least one food material selected from the group consisting of propolis extract, grape skin extract and hawthorn extract, and is a health food containing these. is there. Further, by combining these food materials, further anticancer effect can be obtained. In the present specification, an extract is used as a food material,
Not limited to the extract, for example, if the components contained in the extract such as dry powder of propolis are sufficiently contained and the same effect as the extract used in the present invention can be obtained, It is within the range.

Propolis is a component constituting a honeycomb, and is said to have bacterial growth inhibition and sterilization, fungal sterilization, analgesic effect and wound healing effect. As components contained in propolis, cinnamic acid, coumarinic acid, phenols such as caffeic acid, flavones, flavonols, flavanones, flavonoids such as flavanols, coumarins, aromatic aldehydes, minerals, vitamins, Organic acid etc. are mentioned.

The raw mass of propolis can be taken as it is, but in general, the propolis extract produced by extracting the propolis (raw mass) with ethanol, dewaxing, and filtering is taken. As the extraction solvent, in addition to ethanol, glycerin, water, or a liquefied gas is used, and a supercritical fluid extraction method using these as an extraction solvent can also be used.

Grape skin contains anthocyanins, flavonoids, resveratrol and the like, and is known to have active oxygen scavenging action and platelet aggregation inhibiting action.

[0012] The hawthorn extract is obtained from mature fruits of the hawthorn family Rosaceae, and is known to have mainly a stomachic effect. In addition, the glycoside has a cardiotonic action, and the esters have a vasodilatory action. It is said. Components contained in hawthorn fruits include amygdalin, quercetin, chlorogenic acid, and ursolic acid and tartaric acid as organic acids.

The above food material is preferably used in the form of an extract for the apoptosis inducer of the present invention. Regarding the method for producing each extract, specific examples are shown below, but in addition, food materials may be squeezed to obtain juice, or those extracted using water or a predetermined organic solvent may be used. May be.

When an organic solvent is used for extraction, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol. , Ethyl methyl ketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oil and fat,
Use 1,1,1,2-tetrafluoroethane, etc., which is acceptable for foods. In particular, hot water, hydrous ethanol,
It is more preferable to use hydrous propylene glycol or the like.

The extraction may be carried out by heating extraction,
Supercritical fluid extraction may be performed.

The supercritical fluid extraction is a method of performing extraction using a supercritical fluid which is a fluid in a state of exceeding the gas-liquid critical point (critical temperature, critical pressure) of a substance. In the present embodiment, carbon dioxide is used as the supercritical fluid, but in addition,
Ethane, ethylene, propane, nitrous oxide (laughing gas)
Etc. may be used.

In addition, a substance having a strong affinity with the substance to be extracted may be used.
You may perform supercritical fluid extraction by the entrainer addition method which increases the solubility of a to-be-extracted substance dramatically, or enhances the selectivity of separation by adding about 20 weight%.

In the supercritical outflow extraction process, an extraction step of extracting the target component with the supercritical fluid and a separation step of separating the target component and the supercritical fluid are performed. In the separation process,
Any of extraction separation by pressure change, extraction separation by temperature change, and extraction separation using an adsorbent / absorbent may be performed. Because supercritical extraction can be operated at relatively low temperatures,
The advantage that it can also be applied to substances that deteriorate and decompose at high temperatures,
There is an advantage that the extraction fluid does not remain, a solvent can be circulated and used, a desolvation step and the like can be omitted, and the steps are simplified.

The extraction may be carried out by a liquid carbon dioxide batch method, a liquid carbon dioxide reflux method, a supercritical carbon dioxide reflux method or the like, or a plurality of extraction methods may be combined.
By combining a plurality of extraction methods, it becomes possible to obtain extracts having various compositions.

An apoptosis inducer can be obtained by using at least one selected from these extracts. That is, the selected one or more extracts alone, or the selected one or more extracts and excipients, fillers, binders, thickeners, emulsifiers, colorants, flavors, food additives, It is mixed with a seasoning or the like to give a liquid or solid form, and these extracts are contained to obtain an apoptosis inducer.

The above-mentioned apoptosis inducer may be used as it is, or may be used as an excipient, a filler, a binder, a thickener,
It is used as a health food by blending with an emulsifier, a coloring agent, a flavor, a food additive, a seasoning and the like.

For example, it can be used in a liquid state such as an ampoule, and royal jelly, vitamins, protein, chitosan, lecithin, etc. are added as a nutritional supplement, and a sugar solution and a seasoning are further added to improve the taste. Can be arranged. And these are capsules such as hard capsules, soft capsules, tablets, as needed.
Alternatively, it may be molded as a pill or in the form of powder, granules, candy, and the like. These may be taken as they are, or may be taken after being dissolved in water, hot water, milk or the like, depending on their shape or taste.

[0023]

[Examples] The action of the following food materials as an apoptosis inducer on cancer cells when DNA is used alone
Fragmentation of was evaluated as an index. Propolis extract (water-soluble propolis powder, Morikawa Kenkodou Co., Ltd.). Grape skin extract (Grape skin powder: Lion Corporation). Hawthorn extract (hawthorn extract: Nippon Powder Chemicals Co., Ltd.).

(Evaluation of apoptosis-inducing action on cancer cells) A human gastric cancer cell line, KA, was added to a medium containing the above food materials so that the final concentration was 5 mg / ml.
TO-III was seeded at a cell density of 4 × 10 ⁵ cells / ml and cultured for 3 days. The final concentration refers to the concentration of the food material contained in the medium. The culture medium was RPMI1 containing fetal bovine serum (10%), penicillin G (50 IU / ml), streptomycin (50 μg / ml).
Culture was performed at 37 ° C. in a 5% CO ₂ atmosphere using 640 medium. And after lysing the cultured cells, RNA
The protein was removed and a DNA sample was obtained.

Subsequently, 2 mM ED was used as a running buffer.
40 mM Tris-acetate buffer (pH-containing TA)
7.5), using a 2% agarose gel as an electrophoretic gel, the obtained DNA sample was applied to each well so that the total DNA amount was 2 μg, and electrophoresis was performed.

Then, ethidium bromide staining was further performed to observe the fragmentation of DNA which is an index for inducing apoptosis. The results are shown in Fig. 1.

In the figure, S represents a DNA fragment obtained from a gastric cancer cell obtained by adding a food material to a medium and culturing for 3 days.
C is gastric cancer cell DNA to which no food material is added, and M is a lane on which a DNA molecular weight marker is electrophoresed. Further, S1 is the case where propolis extract is used, S2 is the grape skin extract, and S3 hawthorn extract is used as the food material. The molecular weight of the DNA fragment at the top of the figure is large, and the molecular weight of the DNA fragment becomes smaller toward the bottom.

The results shown in FIG. 1 indicate that DNA fragments having a small molecular weight are found in gastric cancer cells in the medium to which propolis extract, grape skin extract and hawthorn extract are added, whereas those in the medium not containing these extracts. Since high molecular weight DNA is observed in gastric cancer cells, it is understood that addition of these extracts causes DNA fragmentation and induces apoptosis. That is, since the DNA is cut, the same D
Gastric cancer cells eventually die because they do not replicate cells with NA.

[0029]

EFFECT OF THE INVENTION Propolis extract, grape skin extract,
At least one selected from the group consisting of hawthorn extract
Since cancer cell apoptosis can be induced by using one or more kinds of food materials, a cancer cell apoptosis inducer and a health food containing the same can be provided.

[Brief description of drawings]

FIG. 1 is a schematic diagram of an electrophoresis photograph of a DNA fragment of gastric cancer cells.

[Explanation of symbols]

M DNA molecular weight marker C Gastric cancer cell DNA in medium without addition of food materials S1 Gastric cancer cell DNA in medium supplemented with propolis extract
Fragment S2 Gastric cancer cell DNA in medium containing grape skin extract
Fragment S3 Gastric cancer cell DNA fragment in medium containing hawthorn extract

Front page continuation (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 43/00 105 A61P 43/00 105 (72) Inventor Hiroshi Kikuchi 2-19-27 Hakataekimae, Hakata-ku, Fukuoka-shi, Fukuoka No. Toyo Shinyaku Co., Ltd. F term (reference) 4B018 LB10 MD48 MD78 ME08 4C087 AA01 AA02 BB22 CA14 MA01 NA05 NA14 ZB21 ZB26 4C088 AB51 AB56 BA08 BA10 CA06 CA07 CA10 MA07 NA05 NA14 ZB21 ZB26

Claims (3)

[Claims] 1. A propolis extract, a grape skin extract,
At least one selected from the group consisting of hawthorn extract
A cancer cell apoptosis inducer containing at least one food material. 2. The apoptosis inducer according to claim 1, wherein the cancer cell is a gastric cancer cell. 3. A health food containing the apoptosis inducer according to claim 1.