KR101980809B1 - Pharmaceutical composition for prevention and treatment of pancreatic cancer comprising extracts of oat hull - Google Patents

Abstract

The present invention relates to an active ingredient of a pharmaceutical composition or functional food for prevention or treatment comprising oat bark extract having excellent anticancer effect on pancreatic cancer.
Since the pharmaceutical composition for preventing or treating oats bark extract according to the present invention has an excellent anticancer effect on pancreatic cancer and specifically acts on pancreatic cancer, it has low toxicity on normal cells, and therefore is useful as a pharmaceutical composition for prevention or treatment, May be provided as an active ingredient of the food.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for prevention or treatment of pancreatic cancer including oat bark extract,

The present invention relates to a composition for preventing or treating pancreatic cancer comprising oat bark extract, and a functional food for prevention and improvement of pancreatic cancer.

Oats are a biennial plant of the rice field and the scientific name is Avena sativa. It is also called ogre. There is also one that reaches 1m in height at the bottom. The stem is straight and almost hairless, but the nodule has downward hairs. The leaves are 15-30cm long and 6-12㎜ wide, and are broader than wheat and dark green. The leaf sheath is long and the leaves are short and finely divided. Flowers bloom in May-June, and the length is 20-30cm in cones. The small ears are green with a pedestal and contain two small flowers and hang down.

Oat, like other grains, is mainly composed of carbohydrates, but protein is 13 to 14 percent, more than rice, barley and wheat, and 5 to 6 percent is high. However, the amino acid composition of oats is similar to that of rice, with less lysine, tryptophan, threonine, and methionine. Oats are mainly used as food or feed for livestock, but they are peeled and flattened to make oatmeal or cookies. Oats are rich in fiber, lowering cholesterol in the blood and preventing constipation, and it is effective in lowering blood pressure because it is rich in unsaturated fatty acids. Betaglucan, which is one of the components of oats, is a water-soluble dietary fiber, which has improved immune functions and anti-cancer effects in the human body, and prostaglandin has an effect of suppressing the inflammatory reaction (Skendi, A., et al. properties of water soluble beta-glucans from oat cultivars of Avena sativa and Avena bysantina. "Journal of Cereal Science 38.1 (2003): 15-31.). However, research on oat bark has not been conducted.

Pancreatic cancer is an adenocarcinoma of the pancreas that is located in the upper back, connected to the duodenum and adjacent to the spleen. More than 90% of the pancreatic cancer is adenocarcinoma of the pancreatic duct.

There are several types of pancreatic cancer, the most common of which is benign cystic tumors, which include serous cystic tumors, mucinous cystic tumors, papillary papillary mucinous tumors, solid parenchymal tumors, lymphoid epithelial cysts and cystic teratomas. In addition to pancreatic adenocarcinoma and precursor cell carcinoma, which are exocrine tumors, neuroendocrine tumors are also malignant tumors.

Early detection of pancreatic cancer is very difficult because it is rarely seen at the early stage of development, and blood tests for early detection are not available at present. Because of this, pancreatic cancer has a poor prognosis than other cancers. A recurrence usually occurs in 1 to 2 years after surgery, and distant metastasis to the liver or peritoneum or invasion of the tumor in the vicinity of the surgical site forms a new mass. Therefore, even if radical resection is performed, the 2-year survival rate is only 10%, and most patients die within 6 months after diagnosis. Since 1999, more effective anti-cancer drugs have been developed and used for pancreatic cancer, such as gemcitabine, but they do not respond effectively to other drugs. The development of additional anti-pancreatic cancer drugs is in urgent need of development of the third-generation anti-cancer drug, which is the third-generation anti-cancer drug, and it is urgent to develop new anticancer active materials derived from natural substances as well as chemical or immunological drugs. can see.

Under these circumstances, the inventors of the present invention have made efforts to develop a natural-material-based material having anticancer effect on pancreatic cancer, and as a result, confirmed the anticancer effect of oat bark extract and completed the present invention.

 Li, Donghui, et al. "Pancreatic cancer." The Lancet 363.9414 (2004): 1049-1057.

The main object of the present invention is to provide a pharmaceutical composition for preventing or treating pancreatic cancer comprising oat bark extract.

Another object of the present invention is to provide a composition for health functional food for preventing or ameliorating pancreatic cancer comprising oat bark extract.

Another object of the present invention is to provide a method of preventing or treating pancreatic cancer comprising the step of administering an oat bark extract to a subject other than a human.

In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating pancreatic cancer comprising oat bark extract.

Hereinafter, the pharmaceutical composition for prevention or treatment of pancreatic cancer including oat bark extract according to the present invention will be described in detail.

As used herein, the term " oats " is a biennial herbaceous plant of the genus Porphyra, and its scientific name is Avena sativa. The oat bark of the present invention may be a by-product excluding the grain after the processing of oats.

The term " extract " used in the present invention may mean a liquid component obtained by immersing in a solvent at room temperature or under heating for a certain period of time, and a solid component obtained by removing the solvent from the liquid component . The solvent is not limited to one or more solvents selected from the group consisting of water, C ₁ to C ₁₀ alcohols, hexane and mixtures thereof. The solvent may be, but is not limited to, ethanol.

As used herein, the term " pancreatic cancer " refers to a tumor consisting of cancerous cells of the pancreas, an organs located adjacent to the spleen, connected to the duodenum and located in the upper region of the stomach. Examples of such tumors include benign cystic tumors, malignant pancreatic adenocarcinomas, And neuroendocrine tumors, but are not limited thereto.

As used herein, the term " prevention " may mean all actions that inhibit or delay the onset of pancreatic cancer by administering the oat bark extract according to the present invention to the individual.

As used herein, the term " treatment " or " improvement " may refer to any act that causes the composition of the present invention to be administered to a suspected pancreatic cancer susceptible individual such that the symptoms of pancreatic cancer are either improved or benefited.

The pharmaceutical composition of the present invention may increase the expression of the gene CDKN1A, NF-kB, TGF-beta or TNF-alpha, but is not limited thereto.

The pharmaceutical composition of the present invention can be used as a single preparation and can be used as a combined preparation containing a drug known to have an effect of treating a known pancreatic cancer and can be formulated using a pharmaceutically acceptable carrier or excipient, ≪ / RTI > or by intrusion into a multi-dose container.

In addition, if necessary, other conventional additives such as an antioxidant, a buffer and / or a bacteriostatic agent can be added and used. A diluent, a dispersant, a surfactant, a binder, a lubricant, Pills, capsules, granules or tablets, and the like.

In addition, the pharmaceutical composition of the present invention may contain a pharmaceutically effective amount of oat bark extract. The term " pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and is generally in the range of 0.001 to 1000 mg / kg, preferably 0.05 To 200 mg / kg, more preferably 0.1 to 100 mg / kg, may be administered once a day to several times per day. For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the specific composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer an amount that can achieve the maximum effect in a minimal amount without causing side effects, and can be readily determined by those skilled in the art.

The mode of administration of the pharmaceutical composition according to the present invention is not particularly limited and may be conventionally used in the art. As a non-limiting example of such a mode of administration, the compositions may be administered orally or parenterally. The pharmaceutical composition according to the present invention can be manufactured into various formulations according to the intended administration mode.

The frequency of administration of the composition of the present invention is not particularly limited, but it may be administered once a day or divided into several doses.

The concentration of the composition of the present invention is not particularly limited, but it may be 20 μg / ml or more and less than 250 μg / ml.

In another aspect, the present invention provides a health functional food for prevention or improvement of pancreatic cancer comprising oat bark extract.

The pancreatic cancer is as described above. Since the oat bark extract has been used for a long time as a natural substance and proved to be stable, it can be prepared in the form of food which is common but can prevent or improve pancreatic cancer. The solvent is as described above.

Functional food is the same term as food for special health use (FoSHU). It refers to foods that have been processed so that the biomechanical functions are efficiently displayed in addition to the nutritional supply. , The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, rings and the like in order to obtain a useful effect for prevention or improvement of pancreatic cancer.

The food composition of the present invention may further comprise a pharmaceutically acceptable carrier.

There is no particular limitation on the kind of the food to which the composition containing the oat bark extract of the present invention can be added, and examples thereof include various drinks, gums, tea, vitamin complex, and health supplement foods. The food composition may be supplemented with other ingredients which do not interfere with the preventive or improving effect of pancreatic cancer, and the kind thereof is not particularly limited. For example, various herbal medicine extracts, food-acceptable food-aid additives or natural carbohydrates, such as ordinary foods, may be added as an additional ingredient.

The food-aid additive is added to produce a health functional food of each formulation and can be appropriately selected and used by those skilled in the art. For example, various kinds of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, , A stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like, but the kind is not limited by the above examples.

At this time, the content of the extract contained in the food is not particularly limited, but may be 0.01 to 100% by weight, more preferably 1 to 80% by weight based on the total weight of the food composition.

When the food is a beverage, it may be contained in a proportion of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may contain additional ingredients which are commonly used in food compositions and which can improve odor, taste, vision and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid and the like. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn) and copper (Cu) It may also include amino acids such as lysine, tryptophan, cysteine, valine, and the like. In addition, it is also possible to use antiseptics (such as potassium sorbate, sodium benzoate, salicylic acid, and sodium dehydroacetate), disinfectants (such as bleaching powder and highly bleached white powder, sodium hypochlorite), antioxidants (butylhydroxyanilide (BHA), butylhydroxytoluene BHT), etc.), coloring agents (such as tar pigments), coloring agents (such as sodium nitrite and sodium acetic acid), bleaching agents (sodium sulfite), seasoning (such as MSG sodium glutamate), sweeteners (such as hypoglycemia, , Food additives such as flavorings (vanillin, lactones, etc.), swelling agents (alum, potassium hydrogen D-tartrate), emulsifiers, emulsifiers, thickeners, encapsulating agents, gum bases, foam inhibitors, ) Can be added. The additives are selected according to the type of food and used in an appropriate amount.

The health functional food of the present invention can be manufactured by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. Also, unlike general medicine, there is an advantage that there is no side effect that can occur when a medicine is used for a long time by using food as a raw material, and it is excellent in portability.

In another embodiment, the present invention provides a method of preventing or treating pancreatic cancer comprising administering the oat bark extract to a subject other than a human.

As used herein, the term " individual " may refer to any animal, including a human, who has or may have developed pancreatic cancer. The animal may be, but is not limited to, a mammal such as a cow, a horse, a sheep, a pig, a goat, a camel, a nutrient, a dog, a cat,

The preventive or therapeutic method of the present invention may specifically include administering the composition in a pharmaceutically effective amount to a subject suffering from or at risk of developing pancreatic cancer.

The term " administering " as used herein refers to the introduction of a pharmaceutical composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention may be oral or parenteral May be administered via various routes.

Since the pharmaceutical composition for preventing or treating oats bark extract according to the present invention has excellent anticancer effect on pancreatic cancer and specifically acts on pancreatic cancer, it has low toxicity to normal cells, May be provided as an active ingredient of the food.

FIG. 1 shows the anticancer effect according to the concentration and treatment time of the extract of oat bark.
2 shows the anticancer effect according to the concentration and treatment time of oat bark water extract.
FIG. 3 shows the difference in viability of pancreatic cancer cell line and normal cell according to the concentration and treatment time of oat bark extract. HPNE: normal pancreatic cell, MIA PiCa-2: pancreatic cancer cell line
FIG. 4 shows mRNA expression of an anti-cancer gene according to the treatment time of oat bark extract.

Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Example 1: Preparation of oat husk extract

Add 2 L of nucleic acid to 1 kg of oat bark, stir at room temperature for 24 hours, filter with filter paper, discard the filtrate, and repeat the procedure three times. Then, 2 L of alcohol was added to the filtered oat bark, and the mixture was stirred at room temperature for 24 hours. The mixture was filtered through a filter paper, and the filtration was repeated three times. The filtrate was collected and concentrated using a vacuum concentrator to prepare a brown viscous liquid extract do.

Example 2: Preparation of oat husk water extract

2 L of distilled water was added to the oat husks used in the preparation of the alcohol extract, and the mixture was stirred at room temperature for 24 hours, and extracted and filtered. The filtrate was collected three times and concentrated using a vacuum concentrator to prepare a water extract.

Experimental Example 1: Anti-pancreatic cancer effect of oat husk extract

To investigate the anticancer effect of oat bark extract, oat shell extracts were treated at 0, 20, 40, 60, 80, and 100 μg / mL for 24, 48, and 72 hours, respectively, in the pancreatic cancer cell line Miapaca-2 Were measured. As shown in FIG. 1, even at a very low concentration (20 μg / mL), the viability of pancreatic cancer cells was lowered to 50%, which was found to be higher as the concentration increased.

Experimental Example 2: Anti-pancreatic cancer effect of oat husk water extract

In order to confirm the anticancer effect of oat bark water extract, oat bark water extract was treated at 0, 10, 100, 250, 500, and 1000 μg / mL for 24, 48, and 72 hours in Miapaca-2 pancreatic cancer cell line, Were measured. As shown in FIG. 2, the water extract of oat bark was found to have a viability of 80% or more even at a concentration of 200 μg / mL.

Experimental Example  3: Comparison of survival between pancreatic cancer and normal cells by oat bark extract treatment

In order to confirm the anticancer effect and cytotoxicity of the oat bark extract, oat shell extract of Example 1 was injected into human pancreatic normal ductal epithelial (HPNE) cells, which were a pancreatic cancer cell line Miapaca-2 and a control group, at 0, 10, 20, Cell viability was measured after treatment for 24, 48, and 72 hours at 30, 40, 50, 60, 70, 80, 90, and 100 μg / mL. As shown in FIG. 3, it was confirmed that pancreatic cancer cells were specifically toxic with no effect on normal cells at 20 μg / mL or more and 50 μg / mL or less.

Experimental Example 4: Expression of mRNA expressing anti-cancer gene by treatment with oat bark extract

In order to investigate the effect of oat bark extract on the anti - cancer genes, we compared the gene expression levels between the oat bark extract - treated group and the untreated control group.

분석방법으로 하였으며, 사용한 프라이머의 서열은 다음과 같다. Miapaca-2 pancreatic cancer cells were plated on 6-well plates at a density of 2 × 10 ⁵ cells per well and cultured for 24 hours. On the next day, oat bark extract was treated at a concentration of 50 ug / ml and total RNA was extracted from the cells cultured for 24, 48, and 72 hours with the drug. RNA extraction proceeded with the Trizol reagent (Invitrogen, Cat. 15596026) protocol. DNase-free kit (Ambion, Cat. AM1906) was used in the same manner as the protocol to remove DNase and contaminating DNA. CDNA was synthesized with 1 ug total RNA, and the synthesis method was the same as that of SuperScript ll Reverse Transcriptase (Invitrogen, Cat. 18064-022). The expression level of mRNA was confirmed by Real-time PCR and proceeded according to protocol using Power SYBR Green PCR Master Mix (applied biosystem, Cat. 4367659)

The sequence of the primers used was as follows.

CDKN1A Forward: GGAAGACCATGTGGACCTGT Reverse: TAGGGCTTCCTCTTGGAGAA

NF-kB Forward: CATATTTGGGAAGGCCTGAA Reverse: TTTGAAGGTATGGGCCATCT

TGF-β Forward: GGCCTTTCCTGCTTCTCAT Reverse: GTCCTTGCGGAAGTCAATGT

TNF-α Forward: TGCTTGTTCCTCAGCCTCTT Reverse: TGGGCTACAGGCTTGTCACT

As shown in FIG. 4, the expression levels of NF-kB, TGF-β and TNF-α in the extract-treated group were increased over time as compared with the control group.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Pharmaceutical composition for prevention and treatment of          pancreatic cancer comprising extracts of oat hull <130> KPA161255-KR <160> 8 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CDKN1A Forward primer <400> 1 ggaagaccat gtggacctgt 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CDKN1A Reverse primer <400> 2 tagggcttcc tcttggagaa 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NF-kB Forward primer <400> 3 catatttggg aaggcctgaa 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NF-kB Reverse primer <400> 4 tttgaaggta tgggccatct 20 <210> 5 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TGF-beta Forward primer <400> 5 ggcctttcct gcttctcat 19 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TGF-beta Reverse primer <400> 6 gtccttgcgg aagtcaatgt 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha Forward primer <400> 7 tgcttgttcc tcagcctctt 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha Reverse primer <400> 8 tgggctacag gcttgtcact 20

Claims (9)

A pharmaceutical composition for preventing or treating pancreatic cancer, comprising oat bark extract.
The pharmaceutical composition according to claim 1, wherein the oat bark extract is an extract from any one selected from the group consisting of water, C ₁ to C ₁₀ alcohols, hexane, and mixtures thereof.
The pharmaceutical composition according to claim 1, wherein the extract is ethanol.
The pharmaceutical composition according to claim 1, wherein the pancreatic cancer is a pancreatic adenocarcinoma, a pancreatic precursor cell carcinoma, or a pancreatic neuroendocrine tumor.
2. The pharmaceutical composition according to claim 1, wherein the composition increases expression of CDKN1A, NF-kB, TGF-beta and TNF-alpha.
The pharmaceutical composition for preventing or treating pancreatic cancer according to claim 1, wherein the concentration of the oat bark extract is 20 μg / ml or more and less than 250 μg / ml.
A health functional food composition for prevention or improvement of pancreatic cancer comprising oat bark extract.
The health functional food composition according to claim 7, wherein the oat bark extract is an extract from any one selected from the group consisting of water, C ₁ to C ₁₀ alcohols, hexane and mixtures thereof.
A method for preventing or treating pancreatic cancer, comprising administering the pharmaceutical composition of any one of claims 1 to 6 to a subject other than human.

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