KR101695186B1 - Composition for treating, improving or preventing diabetes - Google Patents
Composition for treating, improving or preventing diabetes Download PDFInfo
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- KR101695186B1 KR101695186B1 KR1020160014989A KR20160014989A KR101695186B1 KR 101695186 B1 KR101695186 B1 KR 101695186B1 KR 1020160014989 A KR1020160014989 A KR 1020160014989A KR 20160014989 A KR20160014989 A KR 20160014989A KR 101695186 B1 KR101695186 B1 KR 101695186B1
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- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention relates to a composition for preventing, ameliorating or treating diabetes. More particularly, the present invention relates to a composition for preventing, ameliorating or treating diabetes mellitus, and more particularly, The present invention relates to a composition for preventing, improving or treating diabetes which can be used as a pharmaceutical or food useful for the prevention or treatment of diabetes.
Description
The present invention relates to a composition for preventing, ameliorating or treating diabetes, and more particularly, to a composition for preventing, improving or treating diabetes mellitus comprising fraction (s) of mushroom ginseng as an active ingredient.
Diabetes is the third most dangerous disease worldwide and is the fifth leading cause of death in Korea.
Diabetes is not a single disease but a metabolic disorder that shares features of hyperglycemia. In diabetes, hyperglycemia is caused by insulin secretion, defects in insulin activity, or both.
Diabetes mellitus is divided into type 1 and
Insulin therapy and oral hypoglycemic agents are the most commonly used methods for the treatment of diabetes.
Higher blood sugar levels lead to various complications, so blood glucose should be lowered to prevent complications. The first principle in the treatment of diabetes is to maintain normal blood sugar, which requires insulin to play a normal role.
Orally administered diabetes drugs can be divided into two groups: promoting insulin secretion prior to their action, increasing the activity of insulin, making less glucose in the body, controlling insulin production and promoting the use of glucose. In addition, insulin is administered mainly as an injection. Recently, studies on insulin that is attached to the body have been carried out.
On the other hand, Pleurotus nebrodensis is a kind of avium mushroom called " aiuotari " in Pleurotus mushroom. It has a patent registration under the name of "Great Mushroom", taking advantage of its genetic characteristics different from those of conventional mushroom. The middle part of edible mushroom is 2 ~ 3㎝, the diameter is 10 ~ 15㎝ and it is known to be the largest size. Its nutritional composition is 10 ~ 40 times higher than similar kinds of mushroom. In particular, the mushrooms of the great king of amino acids, vitamins, minerals and contains a large amount. In addition, the Great King Mushroom inhibits the secretion of acycoline, a neurotransmitter in the brain, and contains 18
As described above, various pharmacological effects of the mushroom of Great king mushroom are known, but the study on the mushroom fraction still remains insignificant.
In the present invention, in order to develop a new drug having an antidiabetic effect, the present inventors conducted a study using the fraction of mushroom of Great king. As a result, it was confirmed that the mushroom fraction of mushroom had excellent antidiabetic effect.
Accordingly, the present invention provides a composition for preventing, ameliorating or treating diabetes which is useful for preventing or treating diabetes by significantly inhibiting activation of PTP-1B (Protein-
In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing or treating diabetes comprising the fraction of Pleurotus nebrodensis as an active ingredient.
Preferably, the fraction is a fraction obtained by successively fractionating the mushroom extract obtained by extracting at least one selected from water, an alcohol having 1 to 4 carbon atoms and a mixed solvent thereof with a solvent using chloroform, ethyl acetate and water.
Preferably, the fraction is a chloroform fraction isolated from an extract of Daehwang mushroom ethanol.
Preferably, the fraction is an ethyl acetate fraction isolated from the mushroom ethanol extract.
The above mushroom fraction inhibits PTP-1B activity and promotes insulin secretion.
The present invention also provides a food composition for preventing or improving diabetes comprising the fraction of Pleurotus nebrodensis as an active ingredient.
According to the present invention, it is possible to provide a pharmaceutical or food composition useful for prevention or treatment of diabetes by significantly inhibiting the activation of PTP-1B (Protein-
FIG. 1 is a graph showing the results of measuring the inhibitory activity of? -Glucosidase in the fraction of the mushroom of Great king according to an embodiment of the present invention.
FIG. 2 is a graph showing the results of measurement of PTP-1B (Protein-
FIG. 3 is a graph showing the effect on the viability of RIN-m5f cells of the mushroom fraction according to an embodiment of the present invention.
FIG. 4 is a graph showing an effect on the insulin secretion amount in the cells (RIN-m5f) of the mushroom fraction of Daehangwu according to an embodiment of the present invention.
Hereinafter, the present invention will be described in detail.
The present invention provides a composition for preventing, improving or treating diabetes comprising the fraction of Pleurotus nebrodensis as an active ingredient.
The term "fraction " in the present invention means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents. In the present invention, the term " mushroom extract " means a product obtained by a fractionation method for separating a specific component or a specific group from the mushroom extract. Also, the above-mentioned mushroom extract is a concentrate obtained by squeezing the mushroom of Great King Mushroom with an appropriate liquid and evaporating the liquid. The extract obtained by the extraction treatment, the diluted solution or concentrate of the extract, the dried product obtained by drying the extract, It may be purified water.
It is needless to say that the mushroom can be cultured using a medium used for ordinary mushroom culture.
Specifically, the mushroom may be cultured in a medium containing at least one selected from cottonseed, corn, wheat, rice bran, cottonseed, cone, sugarcane, tangerine husk, sugar, soybean meal and gypsum. Preferably 2 to 8% by weight of cottonseed, 2 to 8% by weight of cottonseed, 10 to 26% by weight of corn flour, 2 to 8% by weight of corn flour, 5 to 15% (V / v) of gypsum, 0.2 to 0.8 wt% of sugar, 0.2 to 0.8 wt% of sugar, 2 to 8 wt% of soybean meal, and 0.5 to 1.5 wt% of gypsum But is not limited thereto.
The mushroom is preferably cultured under conditions of 22 to 25 DEG C, a moisture content (RH) of 60 to 65%, and an acidity (pH) of 5.5 to 6.5
The mushroom fraction of the present invention can be obtained by extracting the mushroom of Great King Mushroom with a solvent, filtering the extracted extract, concentrating it under reduced pressure, and then fractionating it with chloroform, ethyl acetate and water in order.
At this time, the mushroom extract of Dae-Wang may be obtained by extracting and isolating from the nature using extraction and separation methods known in the art. The 'extract' defined in the present invention is extracted from a mushroom of Dae Wang using an appropriate solvent and includes, for example, crude extract, polar solvent-soluble extract or non-polar solvent-extractable extract of Daehwang mushroom.
As a suitable solvent for extracting the extract from the above mushroom, any pharmaceutically acceptable solvent may be used, and water or an organic solvent may be used, but the present invention is not limited thereto. For example, the solvent may be selected from the group consisting of purified water, alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, isopropanol, and butanol, Can be used. Preferably, ethanol (alcohol) is used as the solvent, and 70% (v / v) ethanol (alcohol) is particularly preferably used.
The extraction temperature is preferably 100 DEG C or lower. As the extraction method, a method such as a hot water extraction method, a cold extraction method, a reflux extraction method, a solvent extraction method, a steam distillation method, an ultrasonic extraction method, an elution method, a compression method and the like can be used and more preferably a hot water extraction method or a reflux extraction method is used.
As described above, the extract of Dae-Wang Mushroom can be fractionated from non-polar to polar using an organic solvent to obtain each solvent fraction.
The chloroform, ethylacetate, water or the mixture thereof can be used as a suitable solvent for fractionation of the mushroom fraction. In particular, the chloroform fraction or the ethyl acetate fraction is more effective in inhibiting PTP-1B activity and promoting insulin secretion than other fractions , The fraction of the mushroom of the present invention is more preferably a chloroform fraction or an ethyl acetate fraction and most preferably a chloroform fraction.
After the extract is obtained by using the organic solvent as described above, a liquid substance can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art, or the solvent can be further evaporated, spray dried or lyophilized have.
In addition, the fraction of the mushroom of the king mushroom which is included as an active ingredient in the composition of the present invention may be prepared in powder form by an additional process such as vacuum distillation, freeze-drying, or spray drying, as described above. have. The fractions may also be obtained as further purified fractions using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like .
Therefore, the mushroom fraction used in the present invention is a concept that includes all the fractions obtained in each step of extraction, fractionation or purification, and purified water, a diluted solution thereof, a concentrate or a dried product.
According to one embodiment of the present invention, the mushroom fraction of Dae-Wang has an excellent effect of inhibiting PTP-1B activity and promoting insulin secretion. Accordingly, the mushroom fraction of the present invention can be useful as a pharmaceutical composition and food composition for the prevention, improvement, and treatment of diabetes mellitus.
The present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus comprising the above-described mushroom fraction as an active ingredient.
The mushroom fraction may be contained in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight based on the total weight of the pharmaceutical composition. If the content is less than 0.01% by weight, the efficiency of taking may be poor. If the content is more than 95% by weight, formulation may be difficult.
The pharmaceutical composition of the present invention may be formulated into various forms according to conventional methods. For example, it can be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions and syrups, and can be formulated in the form of external preparations, suppositories, and sterilized injection solutions. However, the composition of the present invention may be most preferably provided in the form of an external preparation for skin. Specifically, it can be used in the form of a skin preparation, such as a liquid preparation, an ointment, a cream, a lotion, a spray, a patch, a gel or an aerosol.
Also, according to each formulation, it may further comprise a pharmaceutically acceptable carrier, excipient and diluent. In addition, it may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, external preparations such as syrups and aerosols, and sterilized injection solutions according to a conventional method, and preferably used in the form of creams, gels, patches, An ointment preparation, an oral preparation, a lotion preparation, a liniment preparation, a pasta preparation or a cataplasma formulation. For example, in the case of an external preparation for skin that is used locally at the site, conventional additives such as a preservative, a solvent for assisting drug penetration, a softening agent for ointment and cream, and the like, May contain conventional carriers. Suitable agents known in the art are preferably, but not limited to, those disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA).
Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the pharmaceutical composition is formulated or formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, , Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. The components may be added to the active ingredient, the Great King mushroom fraction, either independently or in combination.
As used herein, the term "administering" means providing a pharmaceutical composition of the present invention to a subject in any suitable manner.
The present invention relates to pharmaceutical compositions comprising an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, RTI ID = 0.0 > effective < / RTI > amount. It will be apparent to those skilled in the art that the therapeutically effective dose and frequency of administration for the pharmaceutical compositions of the present invention will vary with the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art, and can be readily determined by those skilled in the art and will vary depending upon factors such as the type of disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, 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 concurrently, and the like. The pharmaceutical compositions of the present invention can be administered to a subject in a variety of routes. But are not limited to, intravenous, intraperitoneal, intramuscular, intraarterial, oral, intracardiac, intramedullary, intradermal, transdermal, intestinal, subcutaneous, sublingual or topical administration.
The pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg / kg / day, and may be administered once a day, or divided into several doses.
The present invention also provides a food composition for preventing or improving diabetes mellitus comprising the fraction of mushroom ginseng as an active ingredient.
The mushroom fraction of the present invention can be used as a health functional food, a food additive or a dietary supplement.
When the above-described mushroom fraction is used as a food additive, it can be suitably used according to a conventional method such as adding the mushroom fraction as it is or mixing it with another food or food ingredient.
In addition, the mixing amount of the mushroom fraction may be suitably changed according to the intended use (prevention, health or therapeutic treatment), and it is preferably 0.01 to 95% by weight based on the total weight of the food composition, Preferably 1 to 80% by weight. If the content is less than 0.01% by weight, the efficiency of taking may be lowered. If the content is more than 95% by weight, the rate of increase of the effect of the used amount may be low.
As a specific example, when producing food or beverage, the mushroom fraction of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.
There is no particular limitation on the type of the food, but examples of the food to which the mushroom fraction of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, , Soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes and the like, all of which include health foods in a conventional sense.
When the food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweetening agents such as dextrin and cyclodextrin; Synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.
The food composition of the present invention can be used for various foods, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusters, stabilizers, preservatives, glycerin, Carbonation agents used in beverages, etc., and may include, but is not limited to, natural fruit juices, fruit juice beverages, and flesh for the manufacture of vegetable beverages. These components may be used independently or in combination. The proportion of the above additives is not particularly limited, but is preferably within a range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.
For long-term consumption intended for health and hygiene purposes or health control purposes, the food composition of the present invention has no problem in terms of safety and can be taken for a long period of time.
Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.
Example 1. Preparation of fractionated mushroom of Great king
50 g of dried mushroom ( Pleurotus nebrodensis ) was extracted twice with 80% (w / w) ethanol in a reflux condenser at 80 ° C to obtain 24.11 g (yield: 48.22%) of mushroom extract. 15 g of the thus obtained mushroom extract was dissolved in 150 ml of water and then the chloroform (CHCl 3 ) layer was separated three times. The resulting chloroform layers were combined and concentrated under reduced pressure to obtain 0.17 g (yield: 1.13%) of the fraction of chloroform of Dae-Wang Mushroom in powder form. Ethyl acetate (CH 3 COOC 2 H 5 ) and water (H 2 O) were sequentially added to the remaining water layer and fractionated in the same manner as above. The mixture was concentrated under reduced pressure and lyophilized to obtain 1.19 g (yield: 7.93%) and water fraction (12.23 g, yield: 81.53%).
Example 2. a-glucosidase inhibition
α-glucosidase is an enzyme that induces glucose degradation in the body and is most directly involved in the increase of blood glucose by decomposing the disaccharide into monosaccharides. Therefore, in order to confirm the anti-diabetic function of the fraction of mushroom, the following experiment was conducted.
First, the? -Glucosidase inhibitory activity was measured by applying a nitrophenol assay. 50 μl of 0.3 U / ml yeast α-glucosidase enzyme solution and 100 μl of 10 mM p-nitrophenyl-α-D-glucopyranoside were added to the chloroform fraction, ethyl acetate fraction and
? -glucosidase inhibitory activity (%) = {1- (absorbance of sample-treated group / absorbance of control group)} x 100
As shown in Fig. 1, it was confirmed that the group treated with the Chloroform fraction and the fraction containing the ethyl acetate fraction of manganese chloride had excellent inhibitory effect on the enzyme α-glucosidase.
Example 3 Inhibition of Protein Tyrosine Phosphatase (protein-
When insulin binds to its receptor, it induces glucose metabolism and glycogen synthesis through IRS-1 (Insulin receptor substrate-1) and PI3K (Phosphoinositide 3-kinase) pathway. At this time, the enzyme that interferes with the insulin-receptor binding is PTP-1B, and inhibiting the activation of PTP-1B induces insulin action. Thus, in the present Example, the following experiment was conducted to confirm antidiabetic function through inhibition of PTP-1B of the fraction of Daehwang mushroom.
PTP1B enzyme activity was measured using para-nitrophenyl phosphate (p-NPP) as a substrate. To the buffer solution containing 50 mM citrate buffer (pH 6.0), 0.1 M NaCl, 1 mM EDTA and 1 mM DTT, 20 mM p-NPP, PTP1B (0.05 g) and the ethanol extract of Daehwang mushroom, chloroform fraction, ethyl Acetate and water fractions were added at a concentration of 0.8 and 4 μg / ml, respectively, and incubated at 37 ° C. for 30 minutes. Then, 10 μl of 10 N NaOH was added to terminate the reaction. At this time, guava leaf extract (4 ㎍ / ml) was used as a control. The absorbance of p-nitrophenol was measured at 405 nm. The enzyme inhibition rate (%) was calculated by adding only the enzyme and the substrate to the buffer solution, measuring the value obtained by adding 100% to the enzyme solution, and calculating the value as 0%. The results are shown in Fig.
As shown in FIG. 2, it can be confirmed that the fraction of the mushroom of the invention, which is an active ingredient of the present invention, inhibits the PTP-1B enzyme more effectively in a concentration-dependent manner compared to the extract of the mushroom of the great king. In particular, it was confirmed that the fraction of Chloroform fraction of Dae-Wang Mushroom showed more than 2-fold inhibition of PTP-1B enzyme activity at a concentration of 4 μg / ml compared with other fractions. In addition, it was confirmed that the PTP-1B enzyme inhibitory activity was significantly superior to the control group treated with Guava leaf extract at a concentration of 4 / / ml in the chloroform fraction and the ethyl acetate fraction treated group.
Example 4. Cell viability
In order to analyze the cell viability of the mushroom fraction prepared in Example 1, an experiment was conducted using MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) assay.
Rin-m5F (rat insulinoma β-cell) cells were purchased from ATCC and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin at 37 ° C and 5% CO 2 . The cultured RIN-m5F cells were inoculated into a 24-well plate at a concentration of 3 × 10 5 cells / well and cultured in an incubator for 24 hours. The extracts of the mushroom and its fractions prepared in Example 1 4, 20, and 100 μg / ml, and measured using MTT buffer after 24 hours. The results are shown in Fig.
As shown in FIG. 3, it was confirmed that the cells were survived up to 100 μg / ml in the ethanol extract of Dae-Wang Mushroom, the chloroform fraction, the ethyl acetate fraction and the water fraction.
Example 5. Insulin secretion ability
As a result of the cell viability test of Example 4, the cell viability was excellent at the whole concentration of Dae-Wang mushroom fraction. Therefore, the medium concentration value (4, 20 占 퐂 / ml) Respectively.
First, Rin-m5F (rat insulinoma β-cell) cells were seeded in a 6-well plate at 3.0 × 10 5 per well and cultured in complete medium for 3 days. Krebs-Ringer bicarbonate KRB) buffer. Then, the KRB buffer was added and the mixture was incubated at 37 ° C for 40 minutes. Then, the buffer was removed, and a normal control vehicle (Vehicle), a glucose control (Glucose 16.7 mM), KCL 25 mM, 20 / / ml) were added and incubated for 1 hour. The supernatant was then centrifuged at 4 ° C for 10 min and the supernatant was collected and stored at -20 ° C. The amount of insulin secreted from the cells was measured with an insulin RIA kit, and the concentration of the cell protein in each well was measured to calculate the amount of insulin secreted per each unit of gram protein. The results are shown in FIG.
As shown in FIG. 4, it was confirmed that the insulin secretion level in the group treated with the extract of Chloroform fraction of Dae-Wang mushroom of the present invention was excellent at the concentrations of 4 / / ml and 20 / / ml. Particularly, in the case of the concentration of 20 / / ml, it was confirmed that the treated group of Chloroform fraction of Dae-Wang mushroom showed insulin secretion improved by 1.5 times or more as compared with the group treated with the extract of Dae-Woo mushroom. It was predicted that the chloroform fraction could be effectively used for prevention, improvement and treatment of diabetes.
Formulation Example 1. Preparation of pharmaceutical preparations
Acid production
20 mg of the fraction of Chloroform of Great King Mushroom, 100 mg of lactose and 10 mg of talt were mixed and filled in airtight bags to prepare powders.
Tablet manufacture
10 mg of the chloroform fraction or the ethyl acetate fraction, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed, and tablets were prepared by tableting according to a conventional method for producing tablets.
Capsule preparation
In accordance with the usual preparation method of capsules, 10 mg of water fraction of Chloroform fraction or ethyl acetate fraction of Daehwang mushroom, 3 mg of crystalline cellulose, 14.8 mg of lactose, and 0.2 mg of magnesium stearate were mixed and filled in gelatin capsules to prepare capsules.
Injection manufacturing
(2 mL) per 10 mL of ampoule of chloroform or ethylacetate fraction, 180 mg of mannitol, 2,974 mg of sterilized distilled water for injection, and 26 mg of Na 2 HPO 4 .2H 2 O according to the usual injection preparation method.
Liquid preparation
In accordance with the usual preparation method of the liquid preparation, 20 mg of the fraction of Chloroform or fraction of ethyl acetate, 10 g of isomerized sugar and 5 g of mannitol were added to purified water and dissolved, and the lemon flavor was added in an appropriate amount. Then, purified water was further added thereto, adjusted to a total volume of 100 mL, filled in a brown bottle, and sterilized to prepare a liquid preparation.
Formulation Example 2. Preparation of food preparation
Health food manufacturing
100 mg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 g of vitamin B12, 10 mg of vitamin C, 10 g of biotin, 1.7 mg of nicotinic acid amide, 50 g of folate, 0.5 mg of calcium pantothenate, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of monobasic potassium phosphate, , 90 mg of potassium citrate, 100 mg of calcium carbonate and 24.8 mg of magnesium chloride were mixed and granules were prepared, and a health food was prepared according to a conventional method. At this time, although the composition ratio of the vitamin and mineral mixture is relatively mixed with the ingredient suitable for health food, it may be arbitrarily modified.
Health drink manufacturing
100 g of vitamin C, vitamin E (powder), 100 g of vitamin E, 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 0.2 g of vitamin A, 0.25 g of vitamin B1, 0.3 g of vitamin B2 and a predetermined amount of water were mixed and heated at 85 DEG C for about 1 hour with stirring. The resulting solution was filtered and sterilized in a sterilized 2 L container, . At this time, although the composition ratio of the ingredients suitable for the beverage is comparatively mixed, the mixture ratio may be arbitrarily varied according to the demand, the demanded country, the intended use, and the regional or national preference.
Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.
Claims (7)
Wherein the ethanol extract is an extract of ethanol 70% (v / v).
Wherein the chloroform fraction and the ethyl acetate fraction are contained in an amount of 0.01 to 95% by weight based on the total weight of the pharmaceutical composition.
Wherein the chloroform fraction and the ethyl acetate fraction inhibit PTP-1B activity and promote insulin secretion.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050078860A (en) * | 2004-02-03 | 2005-08-08 | 학교법인 영광학원 | Functional soju containing extracts of mushrooms basidiocarps or mycelia and process for preparation thereof |
| KR20100112205A (en) * | 2001-04-27 | 2010-10-18 | 아지노모토 가부시키가이샤 | Immunopotentiators |
| KR20150119560A (en) * | 2014-04-15 | 2015-10-26 | 대한민국(농촌진흥청장) | Pharmaceutical composition comprising extract of pleurotus cornucopiae and health functional food |
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2016
- 2016-02-05 KR KR1020160014989A patent/KR101695186B1/en active IP Right Grant
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100112205A (en) * | 2001-04-27 | 2010-10-18 | 아지노모토 가부시키가이샤 | Immunopotentiators |
| KR20050078860A (en) * | 2004-02-03 | 2005-08-08 | 학교법인 영광학원 | Functional soju containing extracts of mushrooms basidiocarps or mycelia and process for preparation thereof |
| KR20150119560A (en) * | 2014-04-15 | 2015-10-26 | 대한민국(농촌진흥청장) | Pharmaceutical composition comprising extract of pleurotus cornucopiae and health functional food |
Non-Patent Citations (2)
| Title |
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| Byoung-rai Lee 외 2명, "Hypoglycemic effect of hot-water and ethanol extract of Pleurotus nebrodensis", 2007 한국생물공학회 추계학술대회 및 국제심포지엄 초록집, 제25면, 2007년 발행* * |
| Hypoglycemic effect of hot-water and ethanol extract of Pleurotus nebrodensis, 한국생물공학회, 2007 추계학술대회 및 국제심포지엄. pp.25-25, 2007.10. |
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