WO2021071334A1 - Pharmaceutical composition for preventing and treating type 2 diabetes, containing quercetin-3-o-xyloside - Google Patents
Pharmaceutical composition for preventing and treating type 2 diabetes, containing quercetin-3-o-xyloside Download PDFInfo
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- WO2021071334A1 WO2021071334A1 PCT/KR2020/013854 KR2020013854W WO2021071334A1 WO 2021071334 A1 WO2021071334 A1 WO 2021071334A1 KR 2020013854 W KR2020013854 W KR 2020013854W WO 2021071334 A1 WO2021071334 A1 WO 2021071334A1
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- WIPO (PCT)
- Prior art keywords
- quercetin
- xyloside
- diabetes
- preventing
- pharmaceutical composition
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
Definitions
- the present invention relates to a pharmaceutical composition for preventing and treating diabetes comprising quercetin-3-O-xyloside, and in more detail, quercetin-3-O-xyloside ( Quercetin-3-O-xyloside) reduces insulin resistance in muscle cells induced by insulin-resistance, promotes glucose uptake, and promotes the expression of GLUT-4, a glucose transporter cell membrane protein. By increasing the glucose absorption rate, it was revealed that it has anti-diabetic effect.
- quercetin-3-O-xyloside is more alpha- than Acarbose, a drug for treating type 2 diabetes.
- Diabetes is one of the metabolic diseases, and it is caused by the inability to secrete insulin or to excrete glucose in the urine due to the inability of insulin to function properly. to be. Diabetes is known to be difficult to detect early because there are few initial symptoms, and it is classified into type 1 diabetes and type 2 diabetes.
- Type 1 diabetes is a disease in which the pancreas does not secrete insulin by nature, and is mainly called childhood diabetes. Because it does not secrete insulin by nature, insulin therapy is required.
- type 2 diabetes unlike type 1 diabetes, in which insulin itself is not produced by nature, is due to both environmental and genetic factors, such as obesity and inactive life, which prevents cells from absorbing glucose properly. It is a symptom (DeFronzo, RA et al., 2015).
- Type 2 diabetes has become a major health care problem worldwide, and reported incidence rates are increasing at an alarming rate. Despite improvements in treatment and drug development, treatment is still insufficient, especially because of the associated side effects of most available drugs.
- Type 2 diabetes is closely related to the rapid increase of the obese population due to a decrease in activity due to westernized eating habits and convenience of living, and thus there is a global health problem. The number of deaths from diabetes is gradually increasing along with the increase in the obese population not only in Korea but also worldwide.According to data from the United Nations released by the National Statistical Office of Korea, Korea had 11,575 people in 2010, 11,642 people in 2015, and 2016.
- pancreatic beta cells ⁇ -Cell
- insulin-resistance due to persistent inflammatory response in liver or adipose tissue and impaired glucose metabolism due to excessive fatty acid metabolism
- pancreatic problems including pancreatic beta cells, cells absorb glucose by insulin.
- Antidiabetic drugs that are currently approved by the U.S. Food and Drug Administration and can be used for a long period of time act as metformin (Dunn CJ, Peters et al., 1995) and insulin sensitizer, which inhibits the production of glucose in the liver and improves peripheral insulin sensitivity.
- metformin Dermat CJ, Peters et al., 1995
- insulin sensitizer which inhibits the production of glucose in the liver and improves peripheral insulin sensitivity.
- In addition to Lobeglitazone which increases insulin action in liver, muscle liver and adipocytes, which are target organs of insulin, there are several other species (Lee JH, Noh CK et al., 2015).
- these drugs also have side effects such as obesity, hypoglycemia, rash, liver disorder, gastrointestinal disorder, decreased leukocyte count, anemia, lactic acidosis, kidney dysfunction, decreased insulin, and dehydration.
- Registration Patent 10-2019-0102834 describes a method for preparing a food having anti-obesity and anti-diabetic activity including the deodeok extract. For this reason, attention has been focused on research to develop various antidiabetic drugs that treat or prevent diabetes. Due to the various side effects of diabetes treatment drugs, studies have recently been conducted to develop diabetes treatment drugs using natural products such as compounds isolated from plants, microorganism-derived natural substances, and flavonoids with minimal side effects to the human body.
- quercetin-3-O-xyloside reduces insulin resistance in muscle cells induced by insulin-resistance and promotes glucose uptake.
- GLUT-4 a glucose transporter cell membrane protein
- quercetin-3-O-xyloside is currently 2 It was revealed that it has excellent antidiabetic activity against type 2 diabetes by inhibiting alpha-glucosidase more than two times more effectively than Acarbose, a drug for treating type diabetes.
- quercetin-3-O -A 1:1 (mole: mole) complex of xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside is quercetin-3-O-xylo Seed (Quercetin-3-O-xyloside)
- quercetin-3-O-xyloside quercetin-3-O-xylo Seed
- the present invention is conceived to solve the above problems, the present invention is a composition for preventing and treating diabetes containing flavonoids without various side effects such as cardiovascular action, central action, liver disorder and kidney disorder caused by conventional synthetic pharmaceutical compositions It is to provide a method of manufacturing and a functional food using the same.
- the present invention can provide a pharmaceutical composition for preventing or treating diabetes, including quercetin-3-O-xyloside.
- the pharmaceutical composition may have an activity of inhibiting alpha-glycosidase ( ⁇ -Glucosidase).
- the pharmaceutical composition may increase the intracellular glucose absorption rate of muscle cells.
- the pharmaceutical composition may contain Quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M.
- quercetin-3-O-xyloside and quercetin-3-O-rhamnoside are included.
- quercetin-3-O-rhamnoside Quercetin-3-O-rhamnoside
- the molar ratio of the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and the quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) ( mole ratio) may be 1:10 to 10:1.
- the quercetin-3-O-xyloside and quercetin-3-O-rhamnoside may be included in a concentration of 0.1 to 100 ⁇ M.
- a health functional food composition for preventing or improving diabetes including quercetin-3-O-xyloside.
- the quercetin-3-O-xyloside may be contained in a concentration of 20 to 40 ⁇ M.
- quercetin-3-O-xyloside and quercetin-3-O-rhamnoside are included. It is possible to provide a health functional food composition for preventing or improving diabetes.
- quercetin-3-O-xyloside and quercetin-3-O-rhamnoside are used in diabetic patients. It is possible to provide a method for preventing or treating diabetes, including administering to the patient.
- a pharmaceutical composition comprising quercetin-3-O-xyloside can be used for preventing or treating diabetes.
- quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) comprising
- the use of the pharmaceutical composition for preventing or treating diabetes can be provided.
- the present invention has fewer side effects caused by conventional synthetic pharmaceutical compositions, and alleviates the inhibition of glucose absorption by insulin resistance of muscle cells, and has antidiabetic activity, quercetin-3-O-xyloside (Quercetin-3-O- It provides a pharmaceutical composition for preventing and treating diabetes containing xyloside) as an active ingredient, or a health functional food for preventing and improving anti-diabetes. Furthermore, a pharmaceutical composition for preventing and treating diabetes containing quercetin-3-O-xyloside and quercetin-3-O-rhamnoside or anti Provides health functional foods for preventing and improving diabetes.
- 1 is a chemical structure of Quercetin-3-O-xyloside.
- Figure 2 is a result of measuring ⁇ -Glucosidase inhibitory activity according to the treatment of Quercetin-3-O-xyloside identified in Example 1, a is the result of measuring the ⁇ -Glucosidase inhibitory activity according to the treatment of Quercetin-3-O-xyloside And b is a result of measuring the ⁇ -Glucosidase inhibitory activity according to the treatment of acarbose, and the activity of the ⁇ -glucosidase enzyme is 2 more than that of Acarbose, a drug for treating type 2 diabetes. It is an experimental result showing that the antidiabetic activity against type 2 diabetes is more excellent by inhibiting more than twice as effectively.
- Example 3 is a measurement result of intramuscular viability of Quercetin-3-O-xyloside confirmed in Example 2, where a is a cytotoxicity test result in muscle cells before differentiation, and b is a cytotoxicity test result in muscle cells after differentiation. to be.
- FIG. 4 is a change in glucose uptake in muscle cells by treatment with Quercetin-3-O-xyloside identified in Example 3.
- FIG. 4 is a change in glucose uptake in muscle cells by treatment with Quercetin-3-O-xyloside identified in Example 3.
- FIG. 5 is a result of measuring the IRS-1 protein expression level, AKT protein expression level, and GLUT-4 protein expression level in muscle cells identified in Example 4, respectively, where a is the IRS-1 protein expression level, AKT protein expression level, and GLUT-4 protein expression level was confirmed by Western blot, b is IRS-1 protein expression level, c is AKT protein expression level, and d is GLUT-4 protein expression level. It is one result.
- FIG. 6 is a result of confirming the change in glucose absorption in insulin-resistant muscle cells by treatment with the Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside 1:1 (mole: mole) complex identified in Example 3.
- FIG. 6 is a result of confirming the change in glucose absorption in insulin-resistant muscle cells by treatment with the Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside 1:1 (mole: mole) complex identified in Example 3.
- the present invention seeks to solve the above-described problems by providing a pharmaceutical composition for preventing and treating diabetes, including Quercetin-3-O-xyloside, and a health functional food for preventing and improving diabetes.
- a pharmaceutical composition for preventing and treating diabetes including Quercetin-3-O-xyloside, and a health functional food for preventing and improving diabetes.
- there are no side effects such as obesity, hypoglycemia, rash, liver disorder, gastrointestinal disorder, reduction in white blood cell count, which are side effects that have been seen in conventional pharmaceutical compositions.
- There is an effect of providing a pharmaceutical composition for preventing and treating diabetes that has an effect of increasing the absorption rate of glucose, and a health functional food for preventing and improving diabetes.
- the present invention can provide a pharmaceutical composition for preventing and treating diabetes containing Quercetin-3-O-xyloside.
- Quercetin-3-O-xyloside is known to have antioxidant and anti-inflammatory effects as a flavonoid-based substance.
- the effects of Quercetin-3-O-xyloside, a flavonoid sugar derivative, were confirmed to have a preventive and therapeutic effect on diabetes.
- Quercetin-3-O-xyloside of the present invention is not particularly limited as long as it can be prepared and/or purchased in general, but may be preferably derived from Quercetin.
- the novel physiological activity of Quercetin-3-O-xyloside, derived from Quercetin was found to have antidiabetic and/or therapeutic activity.
- the Quercetin-3-O-xyloside was produced by attaching Xyloside, one of the sugar derivatives, to Quercetin.
- the Quercetin-3-O-xyloside has ⁇ -Glucosidase inhibitory activity, and alpha-glucosidase enzyme activity than acarbose. It was confirmed that the antidiabetic activity against type 2 diabetes was more excellent by inhibiting more than two times more effectively.
- the Quercetin-3-O-xyloside did not affect the cell viability of muscle cells.
- the Quercetin-3-O-xyloside was confirmed the activity of preventing and treating diabetes by increasing the accumulation of glucose in muscle cells.
- the Quercetin-3-O-xyloside was confirmed to reduce the insulin resistance in muscle cells to prevent and treat diabetes.
- the pharmaceutical composition for preventing and treating diabetes of the present invention may exhibit antidiabetic activity by reducing or increasing the expression of one or more proteins in the group consisting of AKT and IRS-1 (Insulin receptor substrate).
- the Quercetin-3-O-xyloside of the present invention exhibits antidiabetic activity by reducing or increasing the expression of AKT and IRS-1.
- the quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside)
- the (mole: mole) complex increased the antidiabetic efficacy through the synergistic effect of further increasing the glucose absorption rate compared to the quercetin-3-O-xyloside single substance.
- Example 6 a 1:1 (mole: mole) complex containing Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside as active ingredients reduces insulin resistance in muscle cells. The activity of preventing and treating diabetes was confirmed.
- the content of the pharmaceutical composition for preventing and treating diabetes is not particularly limited as long as it contains Quercetin-3-O-xyloside, but may preferably be contained in a concentration of 5 to 50 ⁇ M. If Quercetin-3-O-xyloside is contained in a concentration of 5 ⁇ M or less, sufficient antidiabetic activity may not be obtained, and Quercetin-3-O-xyloside in an amount exceeding 100 ⁇ M is included. If so, problems with increased cytotoxicity may arise.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 5 to 50 ⁇ M
- the muscle does not contain Quercetin-3-O-xyloside
- the growth of muscle cells may not be hindered by 0 to 10%
- the concentration of Quercetin-3-O-xyloside is included in a concentration of 5 to 50 ⁇ M, the growth of muscle cells is 0 to It can be reduced to 7.3%.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside at a concentration of 0 to 200 ⁇ M
- the ⁇ -Glucosidase inhibitory activity measurement result may exhibit an IC50 of 0.414 mM.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M
- the muscle cells that induce insulin resistance do not contain Quercetin-3-O-xyloside and
- the intracellular glucose uptake rate of muscle cells can be increased to 50 to 100%, and preferably, when Quercetin-3-O-xyloside is included in a concentration of 20 to 40 ⁇ M, the intracellular glucose uptake rate of muscle cells Can be increased to 61-102%.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M, compared to muscle cells that do not contain Quercetin-3-O-xyloside, The expression of p-AKT in muscle cells can be increased to 96%.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M, compared to muscle cells that do not contain Quercetin-3-O-xyloside, It can reduce the expression of p-IRS-1 (ser) in muscle cells to 49%.
- the pharmaceutical composition for preventing and treating diabetes of the present invention contains a 1:1 (mole: mole) complex containing Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside at a concentration of 10 ⁇ M, respectively .
- Quercetin-3-O-xyloside Compared with Quercetin-3-O-xyloside, it showed the effect of increasing the intracellular glucose uptake rate of muscle cells to 1 to 30%, preferably Quercetin-3-O-xyloside single substance at a concentration of 20 ⁇ M.
- the intracellular glucose absorption rate of muscle cells can be increased to 0 to 49.8%.
- the pharmaceutical composition for preventing and treating diabetes containing Quercetin-3-O-xyloside according to the present invention is oral, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. It can be formulated and used in the form of a dosage form, external preparation, suppository, or sterile injectable solution. Specifically, in the case of formulation, it may be prepared using diluents or excipients such as generally used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
- Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient, such as starch, calcium carbonate, in the Quercetin-3-O-xyloside. ), sucrose (sucrose), lactose (lactose), gelatin, etc. can be prepared by mixing.
- excipients such as starch, calcium carbonate, in the Quercetin-3-O-xyloside.
- sucrose sucrose
- lactose lactose
- gelatin etc.
- lubricants such as magnesium stearate and talc may also be used.
- Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc.
- various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have.
- Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories.
- non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.
- injectable ester such as ethyl oleate
- a base for suppositories witepsol, macrogol, tween61, cacao butter, laurin, glycerogelatin, and the like may be used.
- the pharmaceutical composition for preventing and treating diabetes containing quercetin-3-O-xyloside according to the present invention may vary depending on the age, sex, and weight of the patient, but is generally 0.01 An amount of to 50 mg/kg, preferably 0.1 to 10 mg/kg, may be administered once to several times a day.
- the dosage of the composition containing Quercetin-3-O-xyloside may increase or decrease depending on the route of administration, the degree of disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any way.
- the present invention comprises quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) for the prevention or treatment of diabetes pharmaceutical Compositions can be provided.
- the mole ratio of the quercetin-3-O-xyloside and the quercetin-3-O-rhamnoside is 1:10 to 10; It may be 1, preferably 1:5 to 5:1, and most preferably 1:1.
- the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) may be included in a concentration of 0.1 to 100 ⁇ M, preferably Preferably, it may be included in a concentration of 10 to 50 ⁇ M, and more preferably, it may be included in a concentration of 10 ⁇ M, but is not limited thereto.
- the present invention provides a health functional food for preventing or improving diabetes, including quercetin-3-O-xyloside.
- the quercetin-3-O-xyloside is known to have antioxidant activity as a flavonoid-based substance.
- quercetin-3-O-xyloside, a flavonoid sugar derivative was confirmed to have a preventive and therapeutic effect on diabetes.
- the quercetin-3-O-xyloside of the present invention is not particularly limited as long as it is commonly prepared and/or purchased, but may be preferably derived from quercetin. .
- Example 3 it was confirmed that the quercetin-3-O-xyloside did not affect the cell viability of muscle cells.
- the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) was confirmed the activity of preventing and treating diabetes by increasing the accumulation of glucose in muscle cells.
- the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) was confirmed to reduce the insulin resistance in muscle cells to prevent and treat diabetes.
- the pharmaceutical composition for preventing and treating diabetes of the present invention may exhibit antidiabetic activity by reducing or increasing the expression of one or more proteins in the group consisting of AKT and IRS-1 (Insulin receptor substrate).
- AKT AKT
- IRS-1 Insulin receptor substrate
- the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) of the present invention is p-AKT and p-IRS-1 It was confirmed that the expression of (ser) was decreased or increased to show antidiabetic activity.
- the content of the health functional food for preventing and improving diabetes is not particularly limited as long as it contains Quercetin-3-O-xyloside, but may preferably be included in a concentration of 5 to 50 ⁇ M. If Quercetin-3-O-xyloside is contained in a concentration of 5 ⁇ M or less, sufficient antidiabetic activity may not be obtained, and an amount of Quercetin-3-O-xyloside exceeding the concentration of 100 ⁇ M/ml In the case of containing, a problem with an increase in cytotoxicity may occur. When quercetin-3-O-xyloside is included in a concentration of 5 to 50 ⁇ M, quercetin-3-O-xyloside is included.
- the growth of muscle cells may not be inhibited by 0 to 10%, preferably quercetin-3-O-xyloside (Quercetin-3-O-xyloside) of 5 to 50 ⁇ M.
- quercetin-3-O-xyloside Quercetin-3-O-xyloside
- the growth of muscle cells can be inhibited to 0 to 7.3%.
- the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 0 to 200 ⁇ M
- alpha-glucosidase ( ⁇ - Glucosidase) inhibitory activity measurement result may show an IC50 of 0.414mM.
- the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M, quercetin-3-O- Compared with muscle cells inducing insulin resistance that do not contain xyloside (Quercetin-3-O-xyloside), the intracellular glucose absorption rate of muscle cells can be increased to 50-100%, preferably quercetin- When 3-O-xyloside (Quercetin-3-O-xyloside) is included in a concentration of 20 to 40 ⁇ M, the intracellular glucose uptake rate of muscle cells can be increased to 61 to 102%.
- the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M, Quercetin-3-O- Compared to muscle cells that do not contain xyloside, p-AKT expression in muscle cells can be increased to 96%.
- the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 ⁇ M, quercetin-3-O- Compared with muscle cells that do not contain xyloside (Quercetin-3-O-xyloside), p-IRS-1 (ser) expression in muscle cells can be reduced to 49%.
- the health beverage composition of the present invention is not particularly limited in liquid components except for containing the quercetin-3-O-xyloside as an essential component in the indicated ratio, and various Eggplant flavors or natural carbohydrates, etc. may be contained as additional ingredients.
- natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; Polysaccharides such as dextrin, cyclodextrin; And sugar alcohols such as xylitol, sorbitol, and erythritol.
- natural flavoring agents taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)
- synthetic flavoring agents sacharin, aspartame, etc.
- the health functional food of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid. And salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like.
- the health functional foods of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination.
- the present invention contains quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) for preventing or improving diabetes health function
- a food composition can be provided.
- the present invention can provide a method for preventing or treating diabetes comprising administering quercetin-3-O-xyloside to a diabetic patient.
- the present invention is a diabetes patient comprising the step of administering quercetin-3-O-xyloside and quercetin-3-O-rhamnoside to a diabetic patient It can provide a method of preventing or treating.
- the present invention can provide a use of a pharmaceutical composition comprising quercetin-3-O-xyloside for preventing or treating diabetes.
- the present invention provides a pharmaceutical composition comprising quercetin-3-O-xyloside and quercetin-3-O-rhamnoside for preventing or treating diabetes Can provide a use.
- Quercetin-3-O-xyloside has a Flavone backbone. It has a structure with a xyloside with an O element at the position of carbon 3.
- alpha-glucosidase ( ⁇ -Glucosidase/Saccharomyces, Sigma, USA) was diluted to 0.3 U/mL using 0.2 M phosphate buffer (pH 7.0).
- Substrate para-nitrophenyl glucopyranoside (pNPG, Sigma, USA) was diluted to a concentration of 0.3 mM using the same buffer solution.
- Quercetin-3-O-xyloside is prepared in 0.2 M phosphate buffer (pH 7.0) to concentrations of 0 ⁇ M, 6.25 ⁇ M, 12.5 ⁇ M, 25 ⁇ M, 50 ⁇ M, 100 ⁇ M or 200 ⁇ M. Diluted.
- Acarbose, WAKO, Japan known to have ⁇ -Glucosidase inhibitory activity as a positive control, also 0 ⁇ M, 125 ⁇ M, 250 ⁇ M, 500 ⁇ M, 1000 ⁇ M, 2000 ⁇ M, 3000 ⁇ M in 0.2 M phosphate buffer (pH 7.0). , Diluted to a concentration of 4000 ⁇ M or 8000 ⁇ M. Then, mix 10 ⁇ L of buffer solution, 20 ⁇ L of Quercetin-3-O-xyloside or acarbose sample and 20 ⁇ L of ⁇ -Glucosidase enzyme solution in a 96-well plate, and mix 5% CO2, 37°C incubator ( incubator) for 15 minutes.
- MTT assay methylthiazolyldiphenyl tetrazolium bromide assay was performed to measure the cell viability of quercetin-3-O-xyloside on C2C12 (Mouse myoblast cell line) muscle cells.
- C2C12 Muse myoblast cell line
- MTT assay muscle cells were divided into before and after differentiation induction. Before induction of differentiation, C2C12 muscle cells were dispensed into a 96-well plate at 0.5 ⁇ 104 cells/well, and Dulbecco's Modified Eagle's Medium High glucose (DMEM HG) in a 5% CO2, 37°C incubator for 24 hours.
- DMEM HG Dulbecco's Modified Eagle's Medium High glucose
- DMEM HG medium and Quercetin-3-O-xyloside were treated at different concentrations (0 ⁇ M, 5 ⁇ M, 10 ⁇ M, 20 ⁇ M, 30 ⁇ M, 40 ⁇ M, or 50 ⁇ M) for 24 hours.
- MTT (3-[4,5-dimethyl-thiazol]-2,5-diphenyl-tetrazolium bromide) dissolved in 5 mg/ml in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) was added to each well.
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- DMSO dimethyl sulfoxide
- C2C12 muscle cells were dispensed into a 96-well plate at 0.5 ⁇ 104 cells/well and then in a 5% CO2, 37°C incubator for 24 hours. Cultured in. After incubation, the culture medium was removed, and the medium was changed to Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA) containing 1% penicillin streptomysin and 2% Horse serum (Gibco, USA) for 7 days. Differentiation was induced.
- DMEM HG medium Dulbecco's Modified Eagle's Medium High glucose
- DMEM HG medium and Quercetin-3-O-xyloside were treated at different concentrations (0 ⁇ M, 5 ⁇ M, 10 ⁇ M, 20 ⁇ M, 30 ⁇ M, 40 ⁇ M, or 50 ⁇ M) for 24 hours.
- Dulbecco's C2C12 muscle cells were dispensed into a 96-well black plate at a density of 0.5 ⁇ 104 cells/well and contained 1% penicillin streptomysin and 2% Horse serum (Gibco, USA). Change the medium with Modified Eagle's Medium High Glucose (DMEM HG medium; Hyclone, USA) to differentiate muscle cells for 7 days and add Dexamethasone (Sigma aldrich, USA) to Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA). Insulin-resistance was induced by treatment at a concentration of 1 ⁇ M for 16 hours.
- DMEM HG medium Modified Eagle's Medium High Glucose
- DMEM LG medium Dulbecco's Modified Eagle's Medium Low Glucose
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- Dexamethasone Sigma aldrich, USA
- DMEM GF medium Dulbecco's Modified Eagle's Medium Glucose Free
- Quercetin-3-O-xyloside was added to 0 ⁇ M, 20 ⁇ M, 30 It was treated for 3 hours and 30 minutes at a concentration of ⁇ M or 40 ⁇ M.
- AKT AKT
- IRS-1 Insulin receptor substrate
- GLUT-4 Glucose transporter type
- C2C12 muscle cells were seeded in a 6-well plate at a concentration of 1 ⁇ 10 5 cells/well, and then 1% penicillin streptomysin, 2% Horse serum (Gibco, USA) containing Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA) was used to differentiate the muscle cells for 7 days.
- DMEM HG medium Dulbecco's Modified Eagle's Medium High glucose
- Dexamethasone (Sigma aldrich, USA) was treated in Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA) at a concentration of 1 ⁇ M for 16 hours to induce insulin-resistance. After induction, the culture medium was removed and starvation was performed in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) for 1 hour.
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- Dexamethasone (Sigma aldrich, USA) was added to Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA) to a concentration of 1 ⁇ M, and Quercetin-3-O-xyloside was added to 0 ⁇ M, 20 ⁇ M, 30 It was treated for 3 hours and 30 minutes at a concentration of ⁇ M or 40 ⁇ M. After quercetin-3-O-xyloside treatment, insulin (Sigma aldrich, USA) was treated at a concentration of 1 ⁇ M for 30 minutes.
- Dulbecco's C2C12 muscle cells were dispensed into a 96-well black plate at a density of 0.5 ⁇ 104 cells/well and contained 1% penicillin streptomysin and 2% Horse serum (Gibco, USA). Change the medium with Modified Eagle's Medium High Glucose (DMEM HG medium; Hyclone, USA) to differentiate muscle cells for 7 days and add Dexamethasone (Sigma aldrich, USA) to Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA). Insulin-resistance was induced by treatment at a concentration of 1 ⁇ M for 16 hours.
- DMEM HG medium Modified Eagle's Medium High Glucose
- DMEM LG medium Dulbecco's Modified Eagle's Medium Low Glucose
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- DPBS Dulbecco's Phosphate-Buffered Saline, 1X
- DMEM GF medium Dulbecco's Modified Eagle's Medium Glucose Free
- Quercetin-3-O-xyloside Quercetin-3-O-rhamnoside was treated at a concentration of 20 ⁇ M for 3 hours and 30 minutes.
- the content of 2-NBDG in the cells was measured at 485 nm (exitation) and 535 nm (emission) with a fluorescence spectrophotometer (device manufacturer).
- the measured content of 2-NBDG is shown in Figure 6.
- FIG. 6 when each sample was treated at a concentration of 20 ⁇ M or 10 ⁇ M, when comparing the group treated with Insulin in Dexamethason and the group treated with Insulin in Dexamethason, and the control group treated with the sample, Quercetin-3-O-xyloside It was confirmed that the glucose uptake level of muscle cells treated with and Quercetin-3-O-rhamnoside, respectively, increased with the concentration.
- the glucose uptake level of the complex mixture of Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside at a concentration of 10 ⁇ M and 1:1 (mole: mole) was higher than when each single substance was treated. It could be confirmed that it increased. This is because quercetin-3-O-xyloside and quercetin-3-O-rhamnoside 1:1 (mole: mole) complexes induced insulin resistance more than when quercetin-3-O-xyloside was treated alone. The results show that it has a synergistic effect to further promote glucose absorption in muscle cells.
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Abstract
The present invention relates to a pharmaceutical composition for preventing and treating diabetes, containing quercetin-3-O-xyloside as an active ingredient. More specifically, the present invention demonstrates that: quercetin-3-O-xyloside reduces insulin resistance in insulin resistance induced myocytes, promotes glucose uptake and promotes the expression of GLUT-4, which is a glucose transporter membrane protein, and increases the rate of glucose uptake, thereby having antidiabetic effects; quercetin-3-O-xyloside inhibits α-glucosidase at least twice as effectively as acarbose, which is a current therapeutic agent for type 2 diabetes, thereby having excellent antidiabetic effects on type 2 diabetes; and, compared to a single substance of quercetin-3-O-xyloside, a 1:1 (mole : mole) complex of quercetin-3-O-xyloside and quercetin-3-O-rhamnoside has the synergistic effect of further increasing the rate of glucose uptake, thereby increasing antidiabetic effects. In addition, the present invention relates to a pharmaceutical composition for preventing and treating diabetes, containing the substance.
Description
본 출원은 2019년 10월 11일 출원된 대한민국 특허출원 제10-2019-0125961호를 우선권으로 주장하고, 상기 명세서 전체는 본 출원의 참고문헌이다. This application claims priority to Korean Patent Application No. 10-2019-0125961 filed on October 11, 2019, and the entire specification is a reference to this application.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는 당뇨 예방 및 치료용 약학적 조성물에 관한 것으로서, 보다 상세하게는 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 인슐린 저항성 (Insulin-resistance)이 유도 된 근육세포에서 인슐린 저항성을 감소시키고, 포도당 흡수 (Glucose uptake)를 촉진시키고, 포도당 운반체 세포막 단백질인 GLUT-4의 발현을 촉진시킴으로써 포도당 흡수율을 증가시켜 항당뇨 효능이 있음을 밝히고, 또한, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 현재 2형 당뇨 치료제 약물인 아카보즈 (Acarbose) 보다 알파-글루코시다제 (α-Glucosidase)를 2배 이상 더 효과적으로 억제하여 2형 당뇨에 대한 항당뇨 활성이 우수함을 밝힌 것이며, 나아가, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)와 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 1:1 (mole : mole) 복합물이 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 단일 물질에 비하여 포도당 흡수율을 더욱 증가시키는 상승효과를 통해 항당뇨 효능이 증가한다는 사실을 밝힌 것이며 본 물질을 포함하는 당뇨 예방 및 치료용 약학적 조성물의 발명에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing and treating diabetes comprising quercetin-3-O-xyloside, and in more detail, quercetin-3-O-xyloside ( Quercetin-3-O-xyloside) reduces insulin resistance in muscle cells induced by insulin-resistance, promotes glucose uptake, and promotes the expression of GLUT-4, a glucose transporter cell membrane protein. By increasing the glucose absorption rate, it was revealed that it has anti-diabetic effect. In addition, quercetin-3-O-xyloside is more alpha- than Acarbose, a drug for treating type 2 diabetes. It was revealed that it has excellent antidiabetic activity against type 2 diabetes by more effectively inhibiting glucosidase (α-Glucosidase) more than two times, and furthermore, quercetin-3-O-xyloside (Quercetin-3-O-xyloside) And quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) 1:1 (mole:mole) complex in a single substance quercetin-3-O-xyloside (Quercetin-3-O-xyloside) In comparison, it has been revealed that the anti-diabetic effect is increased through a synergistic effect of further increasing the glucose absorption rate, and it relates to the invention of a pharmaceutical composition for preventing and treating diabetes containing the present substance.
당뇨병은 대사질환 중에 하나로 인슐린이 정상적으로 분비 되지 않거나 인슐린이 역할을 제대로 하지 못하여 포도당을 소변으로 배출하게 되는 것으로 고혈압, 비만, 대사증후군 등과 같은 다른 질병 등과 함께 발병하며 혈중에 포도당 함량이 높은 것이 주된 특징이다. 당뇨병은 초기증상이 거의 없어 조기에 발견하기 힘든 것으로 알려져 있으며 1형 당뇨와 2형 당뇨로 구분 된다. 1형 당뇨의 경우 선천적으로 췌장에서 인슐린을 분비하지 않는 질환으로 주로 소아 당뇨병이라고 불린다. 선천적으로 인슐린을 분비 하지 않기 때문에 인슐린 치료법을 요한다. 그에 반해 제2형 당뇨병은 선천적으로 인슐린 자체가 생성되지 않는 제 1형 당뇨병과 달리 후천적으로 비만, 비활동적인 생활 등의 환경적인 요인과 유전적인 요인 두 가지로 인해 세포가 포도당을 제대로 흡수하지 못하는 증상이다 (DeFronzo, R. A. 등, 2015). Diabetes is one of the metabolic diseases, and it is caused by the inability to secrete insulin or to excrete glucose in the urine due to the inability of insulin to function properly. to be. Diabetes is known to be difficult to detect early because there are few initial symptoms, and it is classified into type 1 diabetes and type 2 diabetes. Type 1 diabetes is a disease in which the pancreas does not secrete insulin by nature, and is mainly called childhood diabetes. Because it does not secrete insulin by nature, insulin therapy is required. On the other hand, type 2 diabetes, unlike type 1 diabetes, in which insulin itself is not produced by nature, is due to both environmental and genetic factors, such as obesity and inactive life, which prevents cells from absorbing glucose properly. It is a symptom (DeFronzo, RA et al., 2015).
제2형 당뇨병(T2DM)은 세계적으로 중대한 건강관리 문제가 되었고, 보고된 발병률은 놀라운 속도로 증가하고 있다. 치료와 신약 개발의 개선에도 불구하고, 특히 대부분의 이용 가능한 약물의 관련 부작용 때문에 치료는 여전히 불충분하다. 제 2형 당뇨는 서구화된 식습관과 생활의 편리화로 인한 활동량 감소로 인한 비만 인구의 급격한 증가와 밀접한 관련이 있고 이에 따른 전 세계적인 보건 문제가 있다. 우리나라 뿐만 아니라 전 세계적으로 비만인구 증가와 함께 현재 당뇨로 인한 사망자 수 자체도 점차 늘어가고 있는 추세이며 한국 통계청에서 발표한 UN에서 조사한 자료에 따르면 우리나라는 2010년 11,575명, 2015년 11,642명, 2016년 11,986명으로 해마다 당뇨로 인한 사망자 수가 증가하고 있으며 그 외에 UN에서 조사한 134개국 중 극히 일부의 국가들만을 제외하고 해마다 당뇨로 인한 사망자 수는 증가하는 추세이다. 제 2형 당뇨는 여러 가지 기작으로 인해 발병한다. 췌장의 베타 세포 (β-Cell)의 감소, 또는 기능 저하로 인한 인슐린 생성 감소, 간 또는 지방조직에서의 지속적인 염증 반응으로 인한 인슐린 저항성(Insulin-Resistance)과 과도한 지방산 대사로 인한 당 대사 장애, 마지막으로 근육조직에서의 인슐린 저항성(Insulin-Resistance)과 인슐린 부족으로 인해 포도당 흡수가 제한되면서 발병한다. 이러한 발병 기작 중에 췌장의 베타세포를 비롯한 췌장의 문제가 아닌 간, 지방 조직, 그리고 근육 조직에서 나타나는 인슐린 저항성 (Insulin-Resistance)을 세포가 띄게 되면 세포가 인슐린에 의해 포도당 (Glucose) 을 세포가 흡수하여야 하는데 그러지 못 하게 되어 혈중에 포도당 (Glucose)가 과도하게 많아지고 이로 인해 다른 합병증 또한 유발하게 된다. 제 2형 당뇨를 치료하는데 알려진 항당뇨 효과가 있는 기존 약물로는 기본적으로 인슐린 (Insulin)을 비롯하여 메트포르민 (Metformin), 로베글리타존 (Lobeglitazone), 엑세나타이드 (Exenatide), 로지글리타존 (Rosiglitazone) 등이 있다. 이러한 항당뇨 약물은 인슐린 분비 또는 감수성 개선, 포도당 생성 및 흡수 또는 배출 등을 통해 혈당감소를 유도하지만, 이러한 효과와 함께 심각한 부작용과 같은 안정성의 문제가 지속적으로 지적되고 있다. 이로 인하여 현재까지 다양한 항당뇨 약물들이 생산되어져 왔지만 심각한 부작용들로 인해 시판이 중지 된 사례로 로지글리타존 (Rosiglitazone)은 2007년 심근경색 위험과 심혈 관계 사망위험을 증가했다는 내용의 연구논문이 발표되면서 결국 2015년 말 시판이 중단되기도 하였다 (Nissen 등 2007). 현재 미국 식품의약국에서 승인되어 장기간 사용이 가능한 항당뇨제는 간에서의 포도당 생성을 억제시키고 말초 인슐린 감수성을 개선시키는 Metformin ( Dunn CJ, Peters 등, 1995)과 인슐린 민감제 (insulin sensitizer)로 작용하여 인슐린의 표적장기인 간, 근육 간 및 지방세포에서 인슐린 작용을 증가시키는 Lobeglitazone 이 외에도 여러 종들이 있다 (Lee JH, Noh CK 등, 2015). 그러나 이 약물들 또한 비만, 저혈당, 발진, 간 장애, 위장 장애, 백혈구 수치 감소, 빈혈, 젖산산증, 콩팥기능 장애, 인슐린 감소, 탈수 등의 부작용들이 보고되었다. Type 2 diabetes (T2DM) has become a major health care problem worldwide, and reported incidence rates are increasing at an alarming rate. Despite improvements in treatment and drug development, treatment is still insufficient, especially because of the associated side effects of most available drugs. Type 2 diabetes is closely related to the rapid increase of the obese population due to a decrease in activity due to westernized eating habits and convenience of living, and thus there is a global health problem. The number of deaths from diabetes is gradually increasing along with the increase in the obese population not only in Korea but also worldwide.According to data from the United Nations released by the National Statistical Office of Korea, Korea had 11,575 people in 2010, 11,642 people in 2015, and 2016. The number of deaths from diabetes is increasing every year to 11,986, and the number of deaths from diabetes is increasing every year except for only a few of the 134 countries surveyed by the United Nations. Type 2 diabetes develops due to several mechanisms. Decreased pancreatic beta cells (β-Cell), or decreased insulin production due to decreased function, insulin-resistance due to persistent inflammatory response in liver or adipose tissue and impaired glucose metabolism due to excessive fatty acid metabolism, last Insulin-Resistance and lack of insulin in the muscle tissue causes the absorption of glucose to be limited. During this pathogenesis, if the cells become insulin-resistance that appears in liver, adipose tissue, and muscle tissue, not pancreatic problems including pancreatic beta cells, cells absorb glucose by insulin. It should be done, but not being able to do so, resulting in excessive increase in glucose in the blood, which causes other complications. Existing drugs that have antidiabetic effects known to treat type 2 diabetes are basically insulin, metformin, lobeglitazone, exenatide, rosiglitazone, etc. There is this. These antidiabetic drugs induce a decrease in blood sugar through insulin secretion or sensitivity improvement, glucose production and absorption or excretion, but stability problems such as serious side effects along with these effects have been continuously pointed out. As a result, various antidiabetic drugs have been produced so far, but the market was stopped due to serious side effects.Rosiglitazone increased the risk of myocardial infarction and cardiovascular death in 2007. The market was discontinued at the end of the year (Nissen et al. 2007). Antidiabetic drugs that are currently approved by the U.S. Food and Drug Administration and can be used for a long period of time act as metformin (Dunn CJ, Peters et al., 1995) and insulin sensitizer, which inhibits the production of glucose in the liver and improves peripheral insulin sensitivity. In addition to Lobeglitazone, which increases insulin action in liver, muscle liver and adipocytes, which are target organs of insulin, there are several other species (Lee JH, Noh CK et al., 2015). However, these drugs also have side effects such as obesity, hypoglycemia, rash, liver disorder, gastrointestinal disorder, decreased leukocyte count, anemia, lactic acidosis, kidney dysfunction, decreased insulin, and dehydration.
여러 부작용들로 인해 최근에 약용식물 (medicinal herb)의 치료효과에 대한 관심이 높아지면서 천연물에 대한 수요가 증대되고 있으며 최근에는 약용식물을 이용한 항 당뇨제의 연구가 활발하다. 이 중 등록특허 10-2019-0102834에는 더덕 추출물을 포함하는 항비만 및 항당뇨 활성을 갖는 식품의 제조방법을 기재하고 있다. 이러한 이유로 당뇨를 치료하거나 예방하는 다양한 항당뇨 약물을 개발하기 위한 연구에 대한 관심이 집중되어 왔다. 당뇨치료제의 다양한 부작용으로 인해 최근에는 인체에 최소한의 부작용을 가지는 식물로부터 분리한 화합물, 미생물 유래 천연물질, 플라보노이드 등 천연물을 이용한 당뇨치료제 개발을 위한 연구가 이루어지고 있다. 다양한 항당뇨 효능이 있는 천연 물질들 중에 주위에서 쉽게 접할 수 있는 식물유래 폴리페놀 중에서 가장 큰 그룹인 플라보노이드 계열의 퀘르세틴 (Quercetin)의 효능은 이미 많은 논문 등을 통해 알려져 있다 ( Chen S 등, 2016). 그러나 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 정제 물질에 대한 항당뇨 효능은 전 세계적으로도 보고 된 바가 없다.Due to various side effects, as interest in the therapeutic effect of medicinal herbs has increased in recent years, the demand for natural products is increasing. Recently, research on antidiabetic drugs using medicinal plants is active. Among them, Registration Patent 10-2019-0102834 describes a method for preparing a food having anti-obesity and anti-diabetic activity including the deodeok extract. For this reason, attention has been focused on research to develop various antidiabetic drugs that treat or prevent diabetes. Due to the various side effects of diabetes treatment drugs, studies have recently been conducted to develop diabetes treatment drugs using natural products such as compounds isolated from plants, microorganism-derived natural substances, and flavonoids with minimal side effects to the human body. Among natural substances with various antidiabetic effects, the efficacy of the flavonoid series Quercetin, the largest group among plant-derived polyphenols that can be easily encountered in the surroundings, is already known through many papers (Chen S et al., 2016). . However, the antidiabetic efficacy of the quercetin-3-O-xyloside purified substance has not been reported worldwide.
본 발명에서는 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 인슐린 저항성 (Insulin-resistance)이 유도 된 근육세포에서 인슐린 저항성을 감소시키고, 포도당 흡수 (Glucose uptake)를 촉진시키고, 포도당 운반체 세포막 단백질인 GLUT-4의 발현을 촉진시킴으로써 포도당 흡수율을 증가시켜 항당뇨 효능이 있음을 밝히고, 또한, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 현재 2형 당뇨 치료제 약물인 아카보즈 (Acarbose) 보다 알파-글루코시다제 (α-Glucosidase)를 2배 이상 더 효과적으로 억제하여 2형 당뇨에 대한 항당뇨 활성이 우수함을 밝힌 것이며, 나아가, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)와 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 1:1 (mole : mole) 복합물이 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 단일 물질에 비하여 포도당 흡수율을 더욱 증가시키는 상승효과를 통해 항당뇨 효능이 증가한다는 사실을 밝힌 것이다. In the present invention, quercetin-3-O-xyloside reduces insulin resistance in muscle cells induced by insulin-resistance and promotes glucose uptake. , By promoting the expression of GLUT-4, a glucose transporter cell membrane protein, it was found that it has antidiabetic efficacy by increasing the glucose absorption rate. In addition, quercetin-3-O-xyloside is currently 2 It was revealed that it has excellent antidiabetic activity against type 2 diabetes by inhibiting alpha-glucosidase more than two times more effectively than Acarbose, a drug for treating type diabetes. Furthermore, quercetin-3-O -A 1:1 (mole: mole) complex of xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside is quercetin-3-O-xylo Seed (Quercetin-3-O-xyloside) Compared to a single substance, it was revealed that the anti-diabetic efficacy was increased through the synergistic effect of further increasing the glucose absorption rate.
본 발명은 상술한 문제를 해결하기 위해 안출된 것으로, 본 발명은 종래 합성 약제 조성물이 유발하는 심혈관작용, 중추작용, 간장 장애 및 신장장애 등 여러 부작용이 없는 플라보노이드를 포함하는 당뇨 예방 및 치료용 조성물과 제조방법 및 이를 이용한 건강기능성 식품을 제공하는 것이다.The present invention is conceived to solve the above problems, the present invention is a composition for preventing and treating diabetes containing flavonoids without various side effects such as cardiovascular action, central action, liver disorder and kidney disorder caused by conventional synthetic pharmaceutical compositions It is to provide a method of manufacturing and a functional food using the same.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는 당뇨병의 예방 또는 치료용 약학적 조성물을 제공할 수 있다. The present invention can provide a pharmaceutical composition for preventing or treating diabetes, including quercetin-3-O-xyloside.
본 발명의 바람직한 일실시예에 따르면, 상기 약학적 조성물은 알파-글리코시다제 (α-Glucosidase)를 억제하는 활성을 가질 수 있다. According to a preferred embodiment of the present invention, the pharmaceutical composition may have an activity of inhibiting alpha-glycosidase (α-Glucosidase).
본 발명의 바람직한 일실시예에 따르면, 상기 약학적 조성물은 근육세포의 세포 내 포도당 흡수율을 증가시킬 수 있다. According to a preferred embodiment of the present invention, the pharmaceutical composition may increase the intracellular glucose absorption rate of muscle cells.
본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 약학적 조성물은 Quercetin-3-O-xyloside을 20 내지 40 μM의 농도로 포함할 수 있다.According to another preferred embodiment of the present invention, the pharmaceutical composition may contain Quercetin-3-O-xyloside in a concentration of 20 to 40 μM.
본 발명의 바람직한 또 다른 일실시예에 따르면, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는, 당뇨병의 예방 또는 치료용 약학적 조성물을 제공할 수 있다. According to another preferred embodiment of the present invention, quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) are included. To, it is possible to provide a pharmaceutical composition for preventing or treating diabetes.
본 발명의 바람직한 일실시예에 따르면, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 몰비(mole ratio)는 1:10 내지 10:1일 수 있다. According to a preferred embodiment of the present invention, the molar ratio of the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and the quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) ( mole ratio) may be 1:10 to 10:1.
본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)를 0.1 내지 100μM 의 농도로 포함할 수 있다.According to another preferred embodiment of the present invention, the quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) It may be included in a concentration of 0.1 to 100 μM.
본 발명의 바람직한 또 다른 일실시예에 따르면, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물을 제공할 수 있다. According to another preferred embodiment of the present invention, it is possible to provide a health functional food composition for preventing or improving diabetes including quercetin-3-O-xyloside.
본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 20 내지 40 μM 의 농도로 포함할 수 있다. According to another preferred embodiment of the present invention, the quercetin-3-O-xyloside may be contained in a concentration of 20 to 40 μM.
본 발명의 바람직한 또 다른 일실시예에 따르면, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물을 제공할 수 있다. According to another preferred embodiment of the present invention, quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) are included. It is possible to provide a health functional food composition for preventing or improving diabetes.
본 발명의 바람직한 또 다른 일실시예에 따르면 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 당뇨병 환자에게 투여하는 단계를 포함하는 당뇨병의 예방 또는 치료방법을 제공할 수 있다. According to another preferred embodiment of the present invention, it is possible to provide a method for preventing or treating diabetes comprising administering quercetin-3-O-xyloside to a diabetic patient. .
본 발명의 바람직한 또 다른 일실시예에 따르면 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 당뇨병 환자에게 투여하는 단계를 포함하는 당뇨병의 예방 또는 치료방법을 제공할 수 있다. According to another preferred embodiment of the present invention, quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) are used in diabetic patients. It is possible to provide a method for preventing or treating diabetes, including administering to the patient.
본 발명의 바람직한 또 다른 일실시예에 따르면 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도를 제공할 수 있다. According to another preferred embodiment of the present invention, a pharmaceutical composition comprising quercetin-3-O-xyloside can be used for preventing or treating diabetes.
본 발명의 바람직한 또 다른 일실시예에 따르면 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도를 제공할 수 있다. According to another preferred embodiment of the present invention, quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) comprising The use of the pharmaceutical composition for preventing or treating diabetes can be provided.
본 발명은 종래 합성 약제 조성물이 유발하는 부작용들이 적으며, 근육세포의 인슐린 저항에 의한 포도당 흡수 저해를 완화시켜, 항당뇨 활성을 가지는 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 유효성분으로 포함하는 당뇨 예방 및 치료용 약학적 조성물 또는 항당뇨 예방 및 개선용 건강기능성 식품을 제공한다. 나아가 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 당뇨 예방 및 치료용 약학적 조성물 또는 항당뇨 예방 및 개선용 건강기능성 식품을 제공한다.The present invention has fewer side effects caused by conventional synthetic pharmaceutical compositions, and alleviates the inhibition of glucose absorption by insulin resistance of muscle cells, and has antidiabetic activity, quercetin-3-O-xyloside (Quercetin-3-O- It provides a pharmaceutical composition for preventing and treating diabetes containing xyloside) as an active ingredient, or a health functional food for preventing and improving anti-diabetes. Furthermore, a pharmaceutical composition for preventing and treating diabetes containing quercetin-3-O-xyloside and quercetin-3-O-rhamnoside or anti Provides health functional foods for preventing and improving diabetes.
도 1은 Quercetin-3-O-xyloside의 화학적 구조이다. 1 is a chemical structure of Quercetin-3-O-xyloside.
도 2는 실시예 1에서 확인한 Quercetin-3-O-xyloside의 처리에 따른 α-Glucosidase 저해 활성 측정 결과로, a는 Quercetin-3-O-xyloside의 처리에 따른 α-Glucosidase 저해 활성을 측정한 결과이고, b는 아카보즈 (Acarbose)의 처리에 따른 α-Glucosidase 저해 활성을 측정한 결과로서, 2형 당뇨 치료제 약물인 아카보즈 (Acarbose) 보다 알파-글루코시다제 (α-Glucosidase) 효소 활성을 2배 이상 더 효과적으로 억제하여 2형 당뇨에 대한 항당뇨 활성이 더 우수함을 보인다는 실험 결과이다. Figure 2 is a result of measuring α-Glucosidase inhibitory activity according to the treatment of Quercetin-3-O-xyloside identified in Example 1, a is the result of measuring the α-Glucosidase inhibitory activity according to the treatment of Quercetin-3-O-xyloside And b is a result of measuring the α-Glucosidase inhibitory activity according to the treatment of acarbose, and the activity of the α-glucosidase enzyme is 2 more than that of Acarbose, a drug for treating type 2 diabetes. It is an experimental result showing that the antidiabetic activity against type 2 diabetes is more excellent by inhibiting more than twice as effectively.
도 3은 실시예 2에서 확인한 Quercetin-3-O-xyloside의 근육세포 내 생존력 측정 결과로서, a는 분화 전 근육세포에서의 세포독성 실험 결과이고, b는 분화 후 근육세포에서의 세포독성 실험 결과이다. 3 is a measurement result of intramuscular viability of Quercetin-3-O-xyloside confirmed in Example 2, where a is a cytotoxicity test result in muscle cells before differentiation, and b is a cytotoxicity test result in muscle cells after differentiation. to be.
도 4는 실시예 3에서 확인한 Quercetin-3-O-xyloside 처리에 의한 근육세포에서의 포도당 흡수 변화이다. 4 is a change in glucose uptake in muscle cells by treatment with Quercetin-3-O-xyloside identified in Example 3. FIG.
도 5는 실시예 4에서 확인한 근육세포에서의 IRS-1 단백질 발현량, AKT 단백질 발현량 및 GLUT-4 단백질 발현량을 각각 측정 결과로서, a는 IRS-1 단백질 발현량, AKT 단백질 발현량 및 GLUT-4 단백질 발현량을 웨스턴 블랏으로 확인한 결과이고, b는 IRS-1 단백질 발현량을 정량한 결과이며, c는 AKT 단백질 발현량을 정량한 결과이고, d는 GLUT-4 단백질 발현량을 정량한 결과이다. 5 is a result of measuring the IRS-1 protein expression level, AKT protein expression level, and GLUT-4 protein expression level in muscle cells identified in Example 4, respectively, where a is the IRS-1 protein expression level, AKT protein expression level, and GLUT-4 protein expression level was confirmed by Western blot, b is IRS-1 protein expression level, c is AKT protein expression level, and d is GLUT-4 protein expression level. It is one result.
도 6은 실시예 3에서 확인한 Quercetin-3-O-xyloside 및 Quercetin-3-O-rhamnoside 1:1 (mole : mole) 복합물 처리에 의한 인슐린 저항성 근육세포에서의 포도당 흡수 변화를 확인한 결과이다. 6 is a result of confirming the change in glucose absorption in insulin-resistant muscle cells by treatment with the Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside 1:1 (mole: mole) complex identified in Example 3. FIG.
이하, 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
상술한 바와 같이 종래 합성 약제는 비만, 저혈당, 발진, 간 장애, 위장 장애, 백혈구 수치 감소 등 여러 부작용이 나타났으며, 매해마다 세계적으로 증가하는 당뇨로 인한 사망자 수에 따라 이에 대한 새로운 대안이 필요한 실정이었다. 이에 본 발명은 Quercetin-3-O-xyloside를 포함하는 당뇨 예방 및 치료용 약학적 조성물 및 당뇨 예방 및 개선용 건강기능성 식품을 제공함으로써 상술한 문제의 해결을 모색하였다. 이를 통해 종래 약학적 조성물에서 나타났던 부작용인 비만, 저혈당, 발진, 간 장애, 위장 장애, 백혈구 수치 감소 등 여러 부작용이 없으면서, 종래 생약제 추출물에서는 확인이 힘들었던 근육세포의 인슐린 저항성을 저해하고 근육세포 내 포도당의 흡수율을 상승시키는 효과가 있는 당뇨 예방 및 치료용 약학적 조성물 및 당뇨 예방 및 개선용 건강기능성 식품을 제공하는 효과가 있다.As described above, conventional synthetic drugs have had various side effects such as obesity, hypoglycemia, rash, liver disorder, gastrointestinal disorder, and reduced white blood cell count, and a new alternative is needed according to the number of deaths from diabetes that increases worldwide every year. It was true. Accordingly, the present invention seeks to solve the above-described problems by providing a pharmaceutical composition for preventing and treating diabetes, including Quercetin-3-O-xyloside, and a health functional food for preventing and improving diabetes. Through this, there are no side effects such as obesity, hypoglycemia, rash, liver disorder, gastrointestinal disorder, reduction in white blood cell count, which are side effects that have been seen in conventional pharmaceutical compositions. There is an effect of providing a pharmaceutical composition for preventing and treating diabetes that has an effect of increasing the absorption rate of glucose, and a health functional food for preventing and improving diabetes.
본 발명은 Quercetin-3-O-xyloside를 포함하는 당뇨 예방 및 치료용 약학적 조성물을 제공할 수 있다. The present invention can provide a pharmaceutical composition for preventing and treating diabetes containing Quercetin-3-O-xyloside.
상기 Quercetin-3-O-xyloside는 플라보노이드 (flavonoid) 계열의 물질로서 항산화작용 및 항 염증효과가 있다고 알려져 있다. 그런데 본 발명에서는 플라보노이드 당 유도체인 Quercetin-3-O-xyloside의 당뇨에 대한 예방 및 치료 효과를 확인하였다. 본 발명의 Quercetin-3-O-xyloside는 통상적으로 제조 및/또는 구매할 수 있는 것이라면 특별히 제한하지 않으나, 바람직하게는 Quercetin으로부터 유래한 것일 수 있다. Quercetin에서 유래된 Quercetin-3-O-xyloside의 신규한 생리활성인 당뇨 예방 및/또는 치료 활성을 발견하였다. 구체적으로 실시예 1에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside는 Quercetin에 당 유도체 중 하나인 Xyloside를 붙여 생성되었다. 또한, 실시예 2에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside는 α-Glucosidase 저해 활성이 존재함을 확인하였으며, 아카보즈 (Acarbose) 보다 알파-글루코시다제 (α-Glucosidase) 효소 활성을 2배 이상 더 효과적으로 억제하여 2형 당뇨에 대한 항당뇨 활성이 더 우수함을 확인하였다. 또한, 실시예 3에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside는 근육세포의 세포 생존력에는 영향을 미치지 않는 것을 확인하였다. 또한, 실시예 4에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside는 근육세포 내의 포도당 축적을 증가 시켜 당뇨를 예방 및 치료하는 활성을 확인하였다. 또한, 실시예 4에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside는 근육세포 내 인슐린 저항성을 감소시켜 당뇨를 예방 및 치료하는 활성을 확인하였다. 나아가, 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 AKT 및 IRS-1 (Insulin receptor substrate)로 이루어진 군 중 1종 이상의 단백질 발현을 감소 또는 증가시켜 항당뇨 활성을 나타낼 수 있다. 구체적으로, 실시예 5 내지 5 및 도면 5 내지 5 에서 확인되는 바와 같이, 본 발명의 Quercetin-3-O-xyloside은 AKT 및 IRS-1의 발현을 감소 또는 증가시켜 항당뇨 활성을 나타내는 것을 확인할 수 있었다. 실시예 6에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)와 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 1:1 (mole : mole) 복합물이 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 단일 물질에 비하여 포도당 흡수율을 더욱 증가시키는 상승효과를 통해 항당뇨 효능이 증가한다는 사실을 확인하였다. 또한, 실시예 6에서 확인되는 바와 같이, 상기 Quercetin-3-O-xyloside와 Quercetin-3-O-rhamnoside를 유효성분으로 포함한 1:1 (mole : mole) 복합물이 근육세포 내 인슐린 저항성을 감소시켜 당뇨를 예방 및 치료하는 활성을 확인하였다.Quercetin-3-O-xyloside is known to have antioxidant and anti-inflammatory effects as a flavonoid-based substance. However, in the present invention, the effects of Quercetin-3-O-xyloside, a flavonoid sugar derivative, were confirmed to have a preventive and therapeutic effect on diabetes. Quercetin-3-O-xyloside of the present invention is not particularly limited as long as it can be prepared and/or purchased in general, but may be preferably derived from Quercetin. The novel physiological activity of Quercetin-3-O-xyloside, derived from Quercetin, was found to have antidiabetic and/or therapeutic activity. Specifically, as confirmed in Example 1, the Quercetin-3-O-xyloside was produced by attaching Xyloside, one of the sugar derivatives, to Quercetin. In addition, as confirmed in Example 2, it was confirmed that the Quercetin-3-O-xyloside has α-Glucosidase inhibitory activity, and alpha-glucosidase enzyme activity than acarbose. It was confirmed that the antidiabetic activity against type 2 diabetes was more excellent by inhibiting more than two times more effectively. In addition, as confirmed in Example 3, it was confirmed that the Quercetin-3-O-xyloside did not affect the cell viability of muscle cells. In addition, as confirmed in Example 4, the Quercetin-3-O-xyloside was confirmed the activity of preventing and treating diabetes by increasing the accumulation of glucose in muscle cells. In addition, as confirmed in Example 4, the Quercetin-3-O-xyloside was confirmed to reduce the insulin resistance in muscle cells to prevent and treat diabetes. Furthermore, the pharmaceutical composition for preventing and treating diabetes of the present invention may exhibit antidiabetic activity by reducing or increasing the expression of one or more proteins in the group consisting of AKT and IRS-1 (Insulin receptor substrate). Specifically, as can be seen in Examples 5 to 5 and Figures 5 to 5, it can be confirmed that the Quercetin-3-O-xyloside of the present invention exhibits antidiabetic activity by reducing or increasing the expression of AKT and IRS-1. there was. As confirmed in Example 6, 1:1 of the quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) It was confirmed that the (mole: mole) complex increased the antidiabetic efficacy through the synergistic effect of further increasing the glucose absorption rate compared to the quercetin-3-O-xyloside single substance. In addition, as confirmed in Example 6, a 1:1 (mole: mole) complex containing Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside as active ingredients reduces insulin resistance in muscle cells. The activity of preventing and treating diabetes was confirmed.
상기 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 포함하는 것이라면 그 함량을 특별히 제한하지 않으나, 바람직하게는 5 내지 50 μM의 농도로 포함할 수 있다. 만약, 5 μM의 농도 이하의 Quercetin-3-O-xyloside를 포함할 경우, 충분한 항당뇨 활성 효과를 볼 수 없을 수 있으며, 100 μM의 농도를 초과하는 양의 Quercetin-3-O-xyloside를 포함할 경우, 세포독성 증가에 대한 문제가 발생할 수 있다. 좀 더 구체적으로 설명하면, 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 5 내지 50 μM의 농도로 포함할 때, Quercetin-3-O-xyloside를 포함하지 않는 근육세포와 비교하여, 근육세포의 성장에 0 내지 10%로 지장을 주지 않을 수 있으며, 바람직하게는 Quercetin-3-O-xyloside를 5 내지 50μM 의 농도로 포함할 때, 근육세포의 성장을 0 내지 7.3%로 감소시킬 수 있다.The content of the pharmaceutical composition for preventing and treating diabetes is not particularly limited as long as it contains Quercetin-3-O-xyloside, but may preferably be contained in a concentration of 5 to 50 μM. If Quercetin-3-O-xyloside is contained in a concentration of 5 μM or less, sufficient antidiabetic activity may not be obtained, and Quercetin-3-O-xyloside in an amount exceeding 100 μM is included. If so, problems with increased cytotoxicity may arise. More specifically, when the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 5 to 50 μM, the muscle does not contain Quercetin-3-O-xyloside Compared to the cells, the growth of muscle cells may not be hindered by 0 to 10%, and when the concentration of Quercetin-3-O-xyloside is included in a concentration of 5 to 50 μM, the growth of muscle cells is 0 to It can be reduced to 7.3%.
또한 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 0 내지 200 μM의 농도로 포함할 때 α-Glucosidase 저해 활성 측정 결과 IC50 0.414 mM을 나타낼 수 있다. In addition, when the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside at a concentration of 0 to 200 μM, the α-Glucosidase inhibitory activity measurement result may exhibit an IC50 of 0.414 mM.
본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 20 내지 40 μM 의 농도로 포함할 때, Quercetin-3-O-xyloside를 포함하지 않는 인슐린 저항성을 유도한 근육세포와 비교하여, 근육세포의 세포 내 포도당 흡수율을 50 내지 100%로 증가시킬 수 있으며, 바람직하게는 Quercetin-3-O-xyloside를 20 내지 40 μM 의 농도로 포함할 때, 근육세포의 세포 내 포도당 흡수율을 61 내지 102%로 증가시킬 수 있다. 나아가, 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 20 내지 40 μM의 농도로 포함할 때, Quercetin-3-O-xyloside를 포함하지 않는 근육세포와 비교하여, 근육세포 내 p-AKT 발현을 96%로 증가시킬 수 있다. 또한, 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside를 20 내지 40 μM의 농도로 포함할 때, Quercetin-3-O-xyloside를 포함하지 않는 근육세포와 비교하여, 근육세포 내 p- IRS-1 (ser) 발현을 49%로 감소시킬 수 있다.When the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 μM, the muscle cells that induce insulin resistance do not contain Quercetin-3-O-xyloside and In comparison, the intracellular glucose uptake rate of muscle cells can be increased to 50 to 100%, and preferably, when Quercetin-3-O-xyloside is included in a concentration of 20 to 40 μM, the intracellular glucose uptake rate of muscle cells Can be increased to 61-102%. Furthermore, when the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 μM, compared to muscle cells that do not contain Quercetin-3-O-xyloside, The expression of p-AKT in muscle cells can be increased to 96%. In addition, when the pharmaceutical composition for preventing and treating diabetes of the present invention contains Quercetin-3-O-xyloside in a concentration of 20 to 40 μM, compared to muscle cells that do not contain Quercetin-3-O-xyloside, It can reduce the expression of p-IRS-1 (ser) in muscle cells to 49%.
또한 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 Quercetin-3-O-xyloside와 Quercetin-3-O-rhamnoside를 각각 10 μM의 농도로 함유한 1:1 (mole : mole) 복합물을 포함할 때, Quercetin-3-O-xyloside와 비교하여, 근육세포의 세포 내 포도당 흡수율을 1 내지 30%로 상승시키는 효과를 보였으며, 바람직하게는 Quercetin-3-O-xyloside 단일물질을 20 μM의 농도로 포함할 때와 비교하여 근육세포의 세포 내 포도당 흡수율을 0 내지 49.8%로 증가시킬 수 있다. In addition, when the pharmaceutical composition for preventing and treating diabetes of the present invention contains a 1:1 (mole: mole) complex containing Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside at a concentration of 10 μM, respectively , Compared with Quercetin-3-O-xyloside, it showed the effect of increasing the intracellular glucose uptake rate of muscle cells to 1 to 30%, preferably Quercetin-3-O-xyloside single substance at a concentration of 20 μM. Compared with the inclusion, the intracellular glucose absorption rate of muscle cells can be increased to 0 to 49.8%.
본 발명에 따른 Quercetin-3-O-xyloside를 포함하는 당뇨 예방 및 치료용 약학적 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 또는 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 상세하게는, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 Quercetin-3-O-xyloside에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose), 락토오스(lactose), 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용될 수 있다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데, 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. The pharmaceutical composition for preventing and treating diabetes containing Quercetin-3-O-xyloside according to the present invention is oral, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. It can be formulated and used in the form of a dosage form, external preparation, suppository, or sterile injectable solution. Specifically, in the case of formulation, it may be prepared using diluents or excipients such as generally used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient, such as starch, calcium carbonate, in the Quercetin-3-O-xyloside. ), sucrose (sucrose), lactose (lactose), gelatin, etc. can be prepared by mixing. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have.
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제 및 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween)61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween61, cacao butter, laurin, glycerogelatin, and the like may be used.
본 발명에 따른 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는 당뇨 예방 및 치료용 약학적 조성물은 환자의 나이, 성별, 체중에 따라 달라질 수 있으나, 일반적으로 0.01 내지 50 ㎎/㎏의 양, 바람직하게는 0.1 내지 10 ㎎/㎏의 양을 일일 1회 내지 수회로 나누어 투여할 수 있다. 또한 Quercetin-3-O-xyloside를 포함하는 조성물의 투여량은 투여경로, 질병의 정도, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The pharmaceutical composition for preventing and treating diabetes containing quercetin-3-O-xyloside according to the present invention may vary depending on the age, sex, and weight of the patient, but is generally 0.01 An amount of to 50 mg/kg, preferably 0.1 to 10 mg/kg, may be administered once to several times a day. In addition, the dosage of the composition containing Quercetin-3-O-xyloside may increase or decrease depending on the route of administration, the degree of disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any way.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 당뇨병의 예방 또는 치료용 약학적 조성물을 제공할 수 있다. The present invention comprises quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) for the prevention or treatment of diabetes pharmaceutical Compositions can be provided.
상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 몰비(mole ratio)는 1:10 내지 10;1일 수 있고, 바람직하게는 1:5 내지 5:1 일 수 있고, 가장 바람직하게는 1:1일 수 있다. The mole ratio of the quercetin-3-O-xyloside and the quercetin-3-O-rhamnoside is 1:10 to 10; It may be 1, preferably 1:5 to 5:1, and most preferably 1:1.
상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)를 0.1 내지 100μM 의 농도로 포함할 수 있으며, 바람직하게는 10 내지 50 μM 의 농도로 포함할 수 있으며, 더 바람직하게는 10μM 의 농도로 포함할 수 있으나, 이에 한정되지는 않는다. The quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) may be included in a concentration of 0.1 to 100 μM, preferably Preferably, it may be included in a concentration of 10 to 50 μM, and more preferably, it may be included in a concentration of 10 μM, but is not limited thereto.
나아가, 본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는 당뇨 예방 또는 개선용 건강기능성 식품을 제공한다. 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 플라보노이드 (flavonoid) 계열의 물질로서 항산화작용이 있다고 알려져 있다. 그런데 본 발명에서는 플라보노이드 당 유도체인 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 당뇨에 대한 예방 및 치료 효과를 확인하였다. Furthermore, the present invention provides a health functional food for preventing or improving diabetes, including quercetin-3-O-xyloside. The quercetin-3-O-xyloside is known to have antioxidant activity as a flavonoid-based substance. However, in the present invention, quercetin-3-O-xyloside, a flavonoid sugar derivative, was confirmed to have a preventive and therapeutic effect on diabetes.
본 발명의 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 통상적으로 제조 및/또는 구매할 수 있는 것이라면 특별히 제한하지 않으나, 바람직하게는 퀘르세틴 (Quercetin)으로부터 유래한 것일 수 있다. 상기 Quercetin에서 유래된 Quercetin-3-O-xyloside의 신규한 생리활성인 당뇨 예방 및/또는 개선 활성을 발견하였다. 구체적으로 실시예 1에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드(Quercetin-3-O-xyloside)는 퀘르세틴 (Quercetin)에 당 유도체 중 하나인 Xyloside를 붙여 생성되었다. 또한, 실시예 2에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 알파-글루코시다제 (α-Glucosidase) 저해 활성이 존재함을 확인하였다. 또한, 실시예 3에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 근육세포의 세포 생존력에는 영향을 미치지 않는 것을 확인하였다. 또한, 실시예 4에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 근육세포 내의 포도당 축적을 증가 시켜 당뇨를 예방 및 치료하는 활성을 확인하였다. 또한, 실시예 4에서 확인되는 바와 같이, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 근육세포 내 인슐린 저항성을 감소시켜 당뇨를 예방 및 치료하는 활성을 확인하였다. 나아가, 본 발명의 당뇨 예방 및 치료용 약학적 조성물은 AKT 및 IRS-1 (Insulin receptor substrate)로 이루어진 군 중 1종 이상의 단백질 발현을 감소 또는 증가시켜 항당뇨 활성을 나타낼 수 있다. 구체적으로, 실시예 5 내지 5 및 도면 5 내지 5 에서 확인되는 바와 같이, 본 발명의 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 p-AKT 및 p-IRS-1 (ser)의 발현을 감소 또는 증가시켜 항당뇨 활성을 나타내는 것을 확인할 수 있었다. The quercetin-3-O-xyloside of the present invention is not particularly limited as long as it is commonly prepared and/or purchased, but may be preferably derived from quercetin. . A novel physiological activity of the Quercetin-derived Quercetin-3-O-xyloside, an activity for preventing and/or improving diabetes, was found. Specifically, as confirmed in Example 1, the quercetin-3-O-xyloside was produced by adding Xyloside, one of the sugar derivatives, to quercetin. In addition, as confirmed in Example 2, it was confirmed that the quercetin-3-O-xyloside has an alpha-glucosidase inhibitory activity. In addition, as confirmed in Example 3, it was confirmed that the quercetin-3-O-xyloside did not affect the cell viability of muscle cells. In addition, as confirmed in Example 4, the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) was confirmed the activity of preventing and treating diabetes by increasing the accumulation of glucose in muscle cells. In addition, as confirmed in Example 4, the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) was confirmed to reduce the insulin resistance in muscle cells to prevent and treat diabetes. Furthermore, the pharmaceutical composition for preventing and treating diabetes of the present invention may exhibit antidiabetic activity by reducing or increasing the expression of one or more proteins in the group consisting of AKT and IRS-1 (Insulin receptor substrate). Specifically, as shown in Examples 5 to 5 and Figures 5 to 5, the quercetin-3-O-xyloside (Quercetin-3-O-xyloside) of the present invention is p-AKT and p-IRS-1 It was confirmed that the expression of (ser) was decreased or increased to show antidiabetic activity.
더불어, 상기 당뇨 예방 및 개선용 건강기능성 식품은 Quercetin-3-O-xyloside를 포함하는 것이라면 그 함량을 특별히 제한하지 않으나, 바람직하게는 5 내지 50 μM의 농도로 포함할 수 있다. 만약, 5 μM 의 농도 이하의 Quercetin-3-O-xyloside를 포함할 경우, 충분한 항당뇨 활성 효과를 볼 수 없을 수 있으며, 100 ㎍/㎖의 농도를 초과하는 양의 Quercetin-3-O-xyloside를 포함할 경우, 세포독성에 증가에 대한 문제가 발생할 수 있다. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 5 내지 50 μM의 농도로 포함할 때, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하지 않는 근육세포와 비교하여, 근육세포의 성장을 0 내지 10%로 억제시키지 않을 수 있으며, 바람직하게는 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 5 내지 50μM 의 농도로 포함할 때, 근육세포의 성장을 0 내지 7.3%로 억제시킬 수 있다.In addition, the content of the health functional food for preventing and improving diabetes is not particularly limited as long as it contains Quercetin-3-O-xyloside, but may preferably be included in a concentration of 5 to 50 μM. If Quercetin-3-O-xyloside is contained in a concentration of 5 μM or less, sufficient antidiabetic activity may not be obtained, and an amount of Quercetin-3-O-xyloside exceeding the concentration of 100 μM/ml In the case of containing, a problem with an increase in cytotoxicity may occur. When quercetin-3-O-xyloside is included in a concentration of 5 to 50 μM, quercetin-3-O-xyloside is included. Compared with the muscle cells that do not, the growth of muscle cells may not be inhibited by 0 to 10%, preferably quercetin-3-O-xyloside (Quercetin-3-O-xyloside) of 5 to 50 μM. When included in a concentration, the growth of muscle cells can be inhibited to 0 to 7.3%.
또한 본 발명의 당뇨 예방 및 개선용 건강기능성 식품은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 0 내지 200 μM의 농도로 포함할 때 알파-글루코시다제 (α-Glucosidase) 저해 활성 측정 결과 IC50 0.414mM을 나타낼 수 있다. 나아가, 본 발명의 당뇨 예방 및 개선용 건강기능성 식품은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 20 ~ 40 μM 의 농도로 포함할 때, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하지 않는 인슐린 저항성을 유도한 근육세포와 비교하여, 근육세포의 세포 내 포도당 흡수율을 50~ 100%로 증가시킬 수 있으며, 바람직하게는 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 20 ~ 40 μM 의 농도로 포함할 때, 근육세포의 세포 내 포도당 흡수율을 61 ~ 102%로 증가시킬 수 있다. 나아가, 본 발명의 당뇨 예방 및 개선용 건강기능성 식품은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 20 ~ 40 μM의 농도로 포함할 때, Quercetin-3-O-xyloside를 포함하지 않는 근육세포와 비교하여, 근육세포 내 p-AKT 발현을 96%로 증가시킬 수 있다. 또한, 본 발명의 당뇨 예방 및 개선용 건강기능성 식품은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 20 ~ 40 μM의 농도로 포함할 때, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하지 않는 근육세포와 비교하여, 근육세포 내 p-IRS-1 (ser) 발현을 49%로 감소시킬 수 있다. In addition, when the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 0 to 200 μM, alpha-glucosidase (α- Glucosidase) inhibitory activity measurement result may show an IC50 of 0.414mM. Furthermore, when the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 μM, quercetin-3-O- Compared with muscle cells inducing insulin resistance that do not contain xyloside (Quercetin-3-O-xyloside), the intracellular glucose absorption rate of muscle cells can be increased to 50-100%, preferably quercetin- When 3-O-xyloside (Quercetin-3-O-xyloside) is included in a concentration of 20 to 40 μM, the intracellular glucose uptake rate of muscle cells can be increased to 61 to 102%. Furthermore, when the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 μM, Quercetin-3-O- Compared to muscle cells that do not contain xyloside, p-AKT expression in muscle cells can be increased to 96%. In addition, when the health functional food for preventing and improving diabetes of the present invention contains quercetin-3-O-xyloside in a concentration of 20 to 40 μM, quercetin-3-O- Compared with muscle cells that do not contain xyloside (Quercetin-3-O-xyloside), p-IRS-1 (ser) expression in muscle cells can be reduced to 49%.
본 발명의 건강 음료 조성물은 지시된 비율로 필수 성분으로서 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 함유하는 외에는 액체성분에는 특별한 제한은 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예로는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린; 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제 (타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등)) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. The health beverage composition of the present invention is not particularly limited in liquid components except for containing the quercetin-3-O-xyloside as an essential component in the indicated ratio, and various Eggplant flavors or natural carbohydrates, etc. may be contained as additional ingredients. Examples of the above-described natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; Polysaccharides such as dextrin, cyclodextrin; And sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have.
상기 외에 본 발명의 건강 기능성 식품은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 건강 기능성 식품들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다.In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid. And salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, the health functional foods of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물을 제공할 수 있다. The present invention contains quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) for preventing or improving diabetes health function A food composition can be provided.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 당뇨병 환자에게 투여하는 단계를 포함하는 당뇨병의 예방 또는 치료방법을 제공할 수 있다. The present invention can provide a method for preventing or treating diabetes comprising administering quercetin-3-O-xyloside to a diabetic patient.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 당뇨병 환자에게 투여하는 단계를 포함하는 당뇨병의 예방 또는 치료방법을 제공할 수 있다. The present invention is a diabetes patient comprising the step of administering quercetin-3-O-xyloside and quercetin-3-O-rhamnoside to a diabetic patient It can provide a method of preventing or treating.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도를 제공할 수 있다. The present invention can provide a use of a pharmaceutical composition comprising quercetin-3-O-xyloside for preventing or treating diabetes.
본 발명은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도를 제공할 수 있다. The present invention provides a pharmaceutical composition comprising quercetin-3-O-xyloside and quercetin-3-O-rhamnoside for preventing or treating diabetes Can provide a use.
이하, 본 발명을 실시예에 의해 상세히 설명하기로 한다. 그러나 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples. However, these examples are for explaining the present invention in more detail, and the scope of the present invention is not limited to these examples.
실시예 1. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)의 화학적 구조Example 1. Chemical structure of quercetin-3-O-xyloside
퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 Flavone 기본 골격을 가진다. 3번 탄소 자리에 O 원소와 함께 자일로시드 (xyloside)를 가진 구조를 띄고 있다.Quercetin-3-O-xyloside has a Flavone backbone. It has a structure with a xyloside with an O element at the position of carbon 3.
실시예 2. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)의 알파-글루코시다제 (α-Glucosidase) 저해 활성 측정 Example 2. Measurement of quercetin-3-O-xyloside's alpha-glucosidase inhibitory activity
알파-글루코시다아제 (α-Glucosidase) 저해 활성 측정을 위해 알파-글루코시다아제 (α-Glucosidase/Saccharomyces, Sigma, USA)는 0.2 M 인산염 완충제(pH 7.0)를 사용하여 0.3 U/mL로 희석하였다. 기질 파라-니트로페닐 글루코피라노사이드 (pNPG, Sigma, USA)는 같은 완충용액을 사용하여 0.3 mM 농도로 희석하였다. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)는 0.2 M 인산염 완충제(pH 7.0)에 0 μM, 6.25μM, 12.5 μM, 25 μM, 50 μM, 100 μM 또는 200 μM 농도까지 희석하였다. Positive control로 α-Glucosidase 억제 활성이 있다고 알려진 아카보즈 (Acarbose, WAKO, Japan) 또한 0.2 M 인산염 완충제(pH 7.0)에 0 μM, 125 μM, 250 μM, 500 μM, 1000 μM, 2000 μM, 3000 μM, 4000 μM 또는 8000μM의 농도로 희석 하였다. 그 후 96-웰 플레이트 (96-well plate)에 완충용액 10 μL, Quercetin-3-O-xyloside 또는 아카보즈 시료 20 μL와 20 μL의 α-Glucosidase 효소액을 혼합하여 5 % CO2, 37℃ 인큐베이터(incubator)에서 15분간 배양하였다. 또한 pNPG와 α-Glucosidase 효소액이 반응하여 흡광도의 차이가 발생할 수 있기에 정확도를 높이고자 Control 군으로 설정하였다. 그리고 시료 자체의 발색으로 인해 흡광도에 차이가 날 수 있으므로 Blank 군을 설정하였다. 그 후 20 μL의 pNPG 용액을 각 샘플과 혼합, Control 및 Blank에 첨가한 후 5 % CO2, 37℃ 인큐베이터 (incubator)에서 15분 동안 다시 배양했다. 이 반응은 0.2 M 탄산나트륨용액 130 μL를 첨가하여 종료하였다. Blank에서 α-Glucosidase 효소 용액은 완충 용액으로 대체하였다. Control에서 시료는 완충 용액으로 대체되었다. 그 후 ELISA reader로 405 ㎚에서 흡광도를 측정하였다. 측정 결과는 하기 도면 2와 같다.측정 결과 IC 50의 경우 현재 시중에서 약으로 쓰이고 있는 아카보즈가 1.155 mM 인데 비해 Quercetin-3-O-xyloside는 약 0.414 mM 의 결과가 나왔으며 α-Glucosidase 저해 활성이 있는 것으로 나타났으며 결과적으로 아카보즈 보다 더 우수한 알파-글루코시다제 억제성을 퀘르세틴-3-O-자일로시드가 가지는 것을 확인 하였다.To measure alpha-glucosidase inhibitory activity, alpha-glucosidase (α-Glucosidase/Saccharomyces, Sigma, USA) was diluted to 0.3 U/mL using 0.2 M phosphate buffer (pH 7.0). . Substrate para-nitrophenyl glucopyranoside (pNPG, Sigma, USA) was diluted to a concentration of 0.3 mM using the same buffer solution. Quercetin-3-O-xyloside is prepared in 0.2 M phosphate buffer (pH 7.0) to concentrations of 0 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM or 200 μM. Diluted. Acarbose, WAKO, Japan, known to have α-Glucosidase inhibitory activity as a positive control, also 0 μM, 125 μM, 250 μM, 500 μM, 1000 μM, 2000 μM, 3000 μM in 0.2 M phosphate buffer (pH 7.0). , Diluted to a concentration of 4000 μM or 8000 μM. Then, mix 10 μL of buffer solution, 20 μL of Quercetin-3-O-xyloside or acarbose sample and 20 μL of α-Glucosidase enzyme solution in a 96-well plate, and mix 5% CO2, 37°C incubator ( incubator) for 15 minutes. In addition, since pNPG and α-Glucosidase enzyme solution reacted, a difference in absorbance may occur, so the control group was set to increase the accuracy. In addition, since the absorbance may vary due to the color development of the sample itself, a blank group was set. Thereafter, 20 μL of pNPG solution was mixed with each sample, added to Control and Blank, and incubated again for 15 minutes in 5% CO2, 37°C incubator. This reaction was terminated by adding 130 μL of 0.2 M sodium carbonate solution. In the blank, the α-Glucosidase enzyme solution was replaced with a buffer solution. In Control, the sample was replaced with a buffer solution. Then, the absorbance was measured at 405 nm with an ELISA reader. The measurement results are shown in Fig. 2. As a result of the measurement, in the case of IC 50, acarbose currently used as a drug on the market was 1.155 mM, whereas the result of Quercetin-3-O-xyloside was about 0.414 mM, and α-Glucosidase inhibitory activity. As a result, it was confirmed that quercetin-3-O-xyloside has better alpha-glucosidase inhibitory properties than acarbose.
|
IC 50 |
시료sample | 결과result |
| 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)Quercetin-3-O-xyloside | 0.414 mM0.414 mM | |
| 아카보즈 (Acarbose)Acarbose | 1.155 mM1.155 mM |
실시예 3. 퀘르세틴-3-O-자일로시드의 근육세포 성장 억제 활성 측정 Example 3. Measurement of the activity of quercetin-3-O-xyloside to inhibit muscle cell growth
C2C12 (Mouse myoblast cell line) 근육세포에 대한 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)의 세포 생존률을 측정하기 위해 MTT 어세이 (methylthiazolyldiphenyl tetrazolium bromide assay)를 실시하였다. MTT 어세이의 경우 근육세포를 분화유도 전과 분화유도 후로 나누어 실시하였다. 분화 유도 전의 경우 C2C12 근육세포를 96-웰 플레이트 (96-well plate)에 0.5×104cells/well로 분주하여 24시간 동안 5 % CO2,37℃인큐베이터 (incubator)에서 Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA)에 10% FBS (Welgene, USA), 1% 페니실린 스트렙토마이신 (penicillin streptomysin)을 첨가한 배지에서 배양하였다. 배양 후 배양액을 제거하고 DMEM HG medium과 Quercetin-3-O-xyloside를 농도별로(0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 또는 50 μM) 24시간 동안 처리하였다. 이후 용매 DPBS (Dulbecco's Phosphate-Buffered Saline, 1X)에 5 ㎎/㎖로 용해한 MTT (3-[4,5-dimethyl-thiazol]-2,5-diphenyl-tetrazolium bromide)용액을 각 웰 (well) 마다 120 ㎕씩 처리하여 20분 동안 5 % CO2, 37℃ 인큐베이터에서 배양하였다. 배양 이후 배양액을 제거한 뒤 각 웰마다 디메틸 설폭사이드 (DMSO; dimethyl sulfoxide) 200 ㎕/well로 넣으면서 피펫을 이용해 잘 교반하였다. 그 후 ELISA reader로 570 ㎚에서 흡광도를 측정하였다. 분화 유도후의 세포생존률 확인은 앞선 분화 유도 전의 경우와 마찬가지로 C2C12 근육세포를 96-웰 플레이트 (96-well plate)에 0.5×104cells/well로 분주하여 24시간 동안 5 % CO2,37℃인큐베이터(incubator)에서 배양하였다. 배양 후 배양액을 제거하고 배지를 1% 페니실린 스트렙토마이신 (penicillin streptomysin), 2% Horse serum (Gibco, USA) 이 함유된 Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA)으로 바꾸어 7일 동안 분화 유도 하였다. 그 후에 DMEM HG medium과 Quercetin-3-O-xyloside를 농도별로(0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 또는 50 μM) 24시간 동안 처리하였다. 이후 용매 DPBS (Dulbecco's Phosphate-Buffered Saline, 1X)에 5 ㎎/㎖로 용해한 MTT (3-[4,5-dimethyl-thiazol]-2,5-diphenyl-tetrazolium bromide)용액을 각 웰 (well) 마다 120 ㎕씩 처리하여 20분 동안 5 % CO2,37℃인큐베이터에서 배양하였다. 배양 이후 배양액을 제거한 뒤 각 웰마다 디메틸 설폭사이드 (DMSO; dimethyl sulfoxide) 200 ㎕/well로 넣으면서 피펫을 이용해 잘 교반하였다. 그 후 ELISA reader로 570 ㎚에서 흡광도를 측정하였다. 도 3에서 확인되는 바와 같이, Quercetin-3-O-xyloside를 0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 또는 50 μM 의 농도로 처리하였을 때, 세포 생존력에는 변화가 없는 것으로 보아 퀘르세틴-3-O-자일로시드는 50 μM 농도까지는 세포 독성은 없는 것으로 나타났다.MTT assay (methylthiazolyldiphenyl tetrazolium bromide assay) was performed to measure the cell viability of quercetin-3-O-xyloside on C2C12 (Mouse myoblast cell line) muscle cells. In the case of the MTT assay, muscle cells were divided into before and after differentiation induction. Before induction of differentiation, C2C12 muscle cells were dispensed into a 96-well plate at 0.5×104 cells/well, and Dulbecco's Modified Eagle's Medium High glucose (DMEM HG) in a 5% CO2, 37°C incubator for 24 hours. medium; Hyclone, USA) was cultured in a medium to which 10% FBS (Welgene, USA) and 1% penicillin streptomysin were added. After incubation, the culture medium was removed, and DMEM HG medium and Quercetin-3-O-xyloside were treated at different concentrations (0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, or 50 μM) for 24 hours. Afterwards, a solution of MTT (3-[4,5-dimethyl-thiazol]-2,5-diphenyl-tetrazolium bromide) dissolved in 5 mg/ml in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) was added to each well. Each 120 µl was treated and incubated in an incubator at 37°C for 20 minutes with 5% CO2. After incubation, the culture medium was removed, and then 200 µl/well of dimethyl sulfoxide (DMSO) was added to each well and stirred well using a pipette. Then, the absorbance was measured at 570 nm with an ELISA reader. To check the cell viability after induction of differentiation, as in the case of the previous induction of differentiation, C2C12 muscle cells were dispensed into a 96-well plate at 0.5×104 cells/well and then in a 5% CO2, 37°C incubator for 24 hours. Cultured in. After incubation, the culture medium was removed, and the medium was changed to Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA) containing 1% penicillin streptomysin and 2% Horse serum (Gibco, USA) for 7 days. Differentiation was induced. Thereafter, DMEM HG medium and Quercetin-3-O-xyloside were treated at different concentrations (0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, or 50 μM) for 24 hours. Afterwards, a solution of MTT (3-[4,5-dimethyl-thiazol]-2,5-diphenyl-tetrazolium bromide) dissolved in 5 mg/ml in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) was added to each well. Each 120 µl was treated and incubated in an incubator at 5% CO2, 37°C for 20 minutes. After incubation, the culture medium was removed, and then 200 µl/well of dimethyl sulfoxide (DMSO) was added to each well and stirred well using a pipette. Then, the absorbance was measured at 570 nm with an ELISA reader. As can be seen in Figure 3, when treated with a concentration of 0 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, or 50 μM of Quercetin-3-O-xyloside, there is no change in cell viability. As a result, quercetin-3-O-xyloside showed no cytotoxicity up to a concentration of 50 μM.
실시예 4. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)가 근육세포에서 포도당 흡수 (Glucose uptake)미치는 영향 조사Example 4. Investigation of the effect of quercetin-3-O-xyloside on glucose uptake in muscle cells
C2C12 근육세포를 96-웰 블랙 플레이트 (96-well black plate)에 0.5 × 104 cells/well 밀도로 분주하고 1% 페니실린 스트렙토마이신 (penicillin streptomysin), 2% Horse serum (Gibco, USA) 이 함유된 Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA)으로 배지를 갈아서 7일 동안 근육세포를 분화시키고 Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA)에 1μM의 농도로 16시간 처리하여 인슐린 저항성 (Insulin-resistance)을 유도 하였다. 유도 후 배양액을 제거 한 뒤 DPBS (Dulbecco's Phosphate-Buffered Saline, 1X)에 1시간 Starvation 시켰다. DPBS를 제거 한 후 Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA)에 1 μM의 농도로 만들고 Quercetin-3-O-xyloside를 0 μM, 20 μM, 30 μM, 또는 40 μM 의 농도로 3시간 30분 처리하였다. Quercetin-3-O-xyloside 처리 이후 insulin (Sigma aldrich, USA)을 1 μM 농도로 30분 처리하였다. 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG : Invitrogen, USA) 을 50 μM 농도로 2시간 처리하였다. 인산완충식염수로 세척한 후 형광분광광도계(기기 제조사)로 485㎚ (exitation), 535㎚ (emission) 에서 세포 내 2-NBDG 함량을 측정하였다. 측정한 2-NBDG 의 함량은 도면 4에 나타냈다. 도 4에서 확인되는 것과 같이, Quercetin-3-O-xyloside를 0 μM, 20 μM, 30 μM, 또는 40 μM 의 농도로 처리한 후 Dexamethason에 Insulin 처리 군과 Dexamethason에 Insulin, 샘플 처리한 대조군을 비교했을 때, Quercetin-3-O-xyloside를 처리한 근육세포의 Glucose uptake 수준이 농도에 따라 증가함을 확인할 수 있었다. 이는 퀘르세틴-3-O-자일로시드가 인슐린 저항성이 유도 된 근육세포에서 포도당 흡수를 촉진 시킨다는 결과를 보여준다.Dulbecco's C2C12 muscle cells were dispensed into a 96-well black plate at a density of 0.5 × 104 cells/well and contained 1% penicillin streptomysin and 2% Horse serum (Gibco, USA). Change the medium with Modified Eagle's Medium High Glucose (DMEM HG medium; Hyclone, USA) to differentiate muscle cells for 7 days and add Dexamethasone (Sigma aldrich, USA) to Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA). Insulin-resistance was induced by treatment at a concentration of 1 μM for 16 hours. After induction, the culture medium was removed and starvation was performed in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) for 1 hour. After removing DPBS, Dexamethasone (Sigma aldrich, USA) was added to Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA) to a concentration of 1 μM, and Quercetin-3-O-xyloside was added to 0 μM, 20 μM, 30 It was treated for 3 hours and 30 minutes at a concentration of μM or 40 μM. After quercetin-3-O-xyloside treatment, insulin (Sigma aldrich, USA) was treated at a concentration of 1 μM for 30 minutes. 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG: Invitrogen, USA) was treated at a concentration of 50 μM for 2 hours. After washing with phosphate buffered saline, the content of 2-NBDG in the cells was measured at 485 nm (exitation) and 535 nm (emission) with a fluorescence spectrophotometer (device manufacturer). The measured content of 2-NBDG is shown in Figure 4. As shown in FIG. 4, after treatment with Quercetin-3-O-xyloside at a concentration of 0 μM, 20 μM, 30 μM, or 40 μM, the Insulin-treated group in Dexamethason and the Insulin in Dexamethason, sample-treated control group were compared. When doing so, it was confirmed that the level of glucose uptake in the muscle cells treated with Quercetin-3-O-xyloside increased with the concentration. This shows that quercetin-3-O-xyloside promotes glucose uptake in muscle cells induced insulin resistance.
실시예 5. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 처리에 의한 근육세포 단백질 발현 기전Example 5. Mechanism of expression of muscle cell protein by quercetin-3-O-xyloside treatment
퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)의 항당뇨 관련 기전을 조사하기 위해 C2C12 근육세포에 대해 AKT, IRS-1 (Insulin receptor substrate), GLUT-4 (Glucose transporter type 4)의 신호 전달에 의한 근육세포의 단백질 발현 감소 또는 증가를 웨스턴 블렛 (Western blot)방법으로 조사하였다. 구체적으로, C2C12 근육세포를 6-웰 플레이트 (6-well plate)에 1 × 105cells/well의 농도로 시딩 (seeding) 후 1% 페니실린 스트렙토마이신 (penicillin streptomysin), 2% Horse serum (Gibco, USA) 이 함유된 Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA)으로 배지를 갈아서 7일 동안 근육세포를 분화시켰다. 그 후Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA)에 1μM의 농도로 16시간 처리하여 인슐린 저항성(Insulin-resistance)을 유도 하였다. 유도 후 배양액을 제거 한 뒤 DPBS (Dulbecco's Phosphate-Buffered Saline, 1X)에 1시간 Starvation 시켰다. DPBS를 제거 한 후 Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA)에 1 μM의 농도로 만들고 Quercetin-3-O-xyloside를 0 μM, 20 μM, 30 μM, 또는 40 μM 의 농도로 3시간 30분 처리하였다. Quercetin-3-O-xyloside 처리 이후 insulin (Sigma aldrich, USA)을 1 μM 농도로 30분 처리하였다. 처리 후 인산완충식염수 (phosphate buffered saline; PBS buffer)로 세척하고 각 웰에 50 ㎕의 용해 버퍼 (lysis buffer)를 첨가 후, 스크래퍼로 세포를 긁어 모은 후 30분 동안 얼음 상에서 반응시키고, 12,000 rpm에서 10분 동안 원심분리 하여 단백질을 수득하였다. 이후 수득한 단백질을 브래드퍼드 분석법 (Bradford 분석법)을 이용하여 단백질을 정량 후 샘플 버퍼(sample buffer)를 넣고 95 ℃에서 5분 동안 변성시킨 뒤에 SDS-PAGE(Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) 겔을 이용하여 전기영동 하였다. 전기영동 후 폴리 비닐 다이플루오라이드 막 (poly-vinyl difluoride membrane)에 옮긴 후, 1차 항-phospho AKT, 항-phospho IRS-1 (Ser), 항-total AKT, 항-total IRS-1, 항-total GLUT-4, 항-베타 액틴 항체 (CST, USA) (1:1000, v/v)를 5% 탈지분유 (skim milk)에 희석하여 폴리 비닐 다이플루오라이드 막에 넣고 4℃에서 24시간 반응시켰다. 이후 TBST (Tris-Buffered Saline와 Tween 20의 혼합액, 0.1% Tween 20)로 10분마다 3번 세척하고 HRP (horeseradish peroxidase)-conjugated 2차 항체를 5% 탈지분유에 1:2000(v/v)으로 희석하여 5 ㎕ 첨가한 후 25 ℃에서 2시간 동안 반응시켰다. 위와 같은 방법으로 TBST를 이용하여 3차례 세척한 후 ECL (enchanced chemiluminescence) 기질 용액을 막아 1분 동안 반응시킨 후에 X-ray 필름에 현상하였다. 얻어진 결과를 이미지 J 프로그램을 이용하여 수치화하였고, 도면 5에 나타냈다. 도 5에서 확인되는 것과 같이, 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 처리한 후 Dexamethason에 Insulin 처리 군과 비교했을 때, Quercetin-3-O-xyloside를 처리한 근육세포에서의 p-IRS-1 (Ser)의 발현량이 감소하는 것을 볼 수 있으며 이로 인해 인슐린 저항성을 퀘르세틴-3-O-자일로시드가 억제 하는 효능이 있음을 알 수 있다. 또한 p-AKT와 포도당 수송 막 단백질인 GLUT-4의 발현량이 증가하는 것을 보아 퀘르세틴-3-O-자일로시드가 근육세포에서 포도당 흡수율 증가에 효능이 있음을 알 수 있는 결과를 보여준다.To investigate the antidiabetic mechanism of quercetin-3-O-xyloside, AKT, IRS-1 (Insulin receptor substrate), GLUT-4 (Glucose transporter type) for C2C12 muscle cells. The decrease or increase of protein expression in muscle cells by the signal transduction of 4) was investigated by the Western blot method. Specifically, C2C12 muscle cells were seeded in a 6-well plate at a concentration of 1 × 10 5 cells/well, and then 1% penicillin streptomysin, 2% Horse serum (Gibco, USA) containing Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA) was used to differentiate the muscle cells for 7 days. Then, Dexamethasone (Sigma aldrich, USA) was treated in Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA) at a concentration of 1 μM for 16 hours to induce insulin-resistance. After induction, the culture medium was removed and starvation was performed in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) for 1 hour. After removing DPBS, Dexamethasone (Sigma aldrich, USA) was added to Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA) to a concentration of 1 μM, and Quercetin-3-O-xyloside was added to 0 μM, 20 μM, 30 It was treated for 3 hours and 30 minutes at a concentration of μM or 40 μM. After quercetin-3-O-xyloside treatment, insulin (Sigma aldrich, USA) was treated at a concentration of 1 μM for 30 minutes. After treatment, wash with phosphate buffered saline (PBS buffer), add 50 µl of lysis buffer to each well, scrape the cells with a scraper and react on ice for 30 minutes, and then react at 12,000 rpm. Protein was obtained by centrifugation for 10 minutes. Subsequently, the obtained protein was quantified using the Bradford method, and then a sample buffer was added and denatured at 95° C. for 5 minutes, followed by SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) gel. Using electrophoresis. After electrophoresis, transfer to a poly-vinyl difluoride membrane, and then the first anti-phospho AKT, anti-phospho IRS-1 (Ser), anti-total AKT, anti-total IRS-1, anti -total GLUT-4, anti-beta actin antibody (CST, USA) (1:1000, v/v) was diluted in 5% skim milk and placed in a polyvinyl difluoride membrane for 24 hours at 4°C. Reacted. Afterwards, wash 3 times every 10 minutes with TBST (a mixture of Tris-Buffered Saline and Tween 20, 0.1% Tween 20) and HRP (horeseradish peroxidase)-conjugated secondary antibody 1:2000 (v/v) in 5% skim milk powder. After diluting and adding 5 µl, the mixture was reacted at 25° C. for 2 hours. After washing three times using TBST in the same manner as described above, the ECL (enchanced chemiluminescence) substrate solution was blocked, reacted for 1 minute, and then developed on an X-ray film. The obtained results were digitized using the Image J program, and are shown in Fig. 5. As shown in Figure 5, when compared to the Insulin-treated group in Dexamethason after treatment with Quercetin-3-O-xyloside, Quercetin-3-O-xyloside was treated It can be seen that the expression level of p-IRS-1 (Ser) in muscle cells decreases, and it can be seen that quercetin-3-O-xyloside has the effect of inhibiting insulin resistance. In addition, as the expression levels of p-AKT and glucose transport membrane protein GLUT-4 are increased, it can be seen that quercetin-3-O-xyloside is effective in increasing the glucose uptake rate in muscle cells.
실시예 6. 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)와 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnloside)의 1:1 (mole : mole) 복합물이 인슐린 저항성이 유도된 근육세포에서 포도당 흡수 (Glucose uptake) 상승에 미치는 영향 조사Example 6.Quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnloside) 1:1 (mole: mole) complex Investigation of the effect of insulin resistance on the increase of glucose uptake in induced muscle cells
C2C12 근육세포를 96-웰 블랙 플레이트 (96-well black plate)에 0.5 × 104 cells/well 밀도로 분주하고 1% 페니실린 스트렙토마이신 (penicillin streptomysin), 2% Horse serum (Gibco, USA) 이 함유된 Dulbecco's Modified Eagle's Medium High glucose (DMEM HG medium; Hyclone, USA)으로 배지를 갈아서 7일 동안 근육세포를 분화시키고 Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA)에 1μM의 농도로 16시간 처리하여 인슐린 저항성 (Insulin-resistance)을 유도 하였다. 유도 후 배양액을 제거 한 뒤 DPBS (Dulbecco's Phosphate-Buffered Saline, 1X)에 1시간 Starvation 시켰다. DPBS를 제거 한 후 Dexamethasone (Sigma aldrich, USA)을 Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA)에 1 μM의 농도로 만들고 Quercetin-3-O-xyloside, Quercetin-3-O-rhamnoside를 20 μM 농도로 3시간 30분 처리하였다. 또한 Quercetin-3-O-xyloside와 Quercetin-3-O-rhamnoside를 각각 10 μM 농도로 1:1 (mole : mole) Mixture를 제조하여 3시간 30분 처리하였다. Quercetin-3-O-xyloside, Quercetin-3-O-rhamnoside, Mixture 처리 이후 insulin (Sigma aldrich, USA)을 1 μM 농도로 30분 처리하였다. 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG : Invitrogen, USA) 을 50 μM 농도로 2시간 처리하였다. 인산완충식염수로 세척한 후 형광분광광도계(기기 제조사)로 485 nm (exitation), 535 nm (emission) 에서 세포 내 2-NBDG 함량을 측정하였다. 측정한 2-NBDG 의 함량은 도면 6에 나타냈다. 도 6에서 확인되는 것과 같이, 각각의 샘플을 20 μM, 또는 10 μM 의 농도로 처리한 후 Dexamethason에 Insulin 처리 군과 Dexamethason에 Insulin, 샘플 처리한 대조군을 비교했을 때, Quercetin-3-O-xyloside와 Quercetin-3-O-rhamnoside를 각각 처리한 근육세포의 Glucose uptake 수준이 농도에 따라 증가함을 확인할 수 있었다. 또한 Quercetin-3-O-xyloside와 Quercetin-3-O-rhamnoside를 각각 10 μM 농도로 1:1 (mole : mole)로 섞은 복합물의 Glucose uptake 수준이 단일물질을 각각 처리했을 때보다 포도당 흡수율이 더욱 증가함을 확인할 수 있었다. 이는 퀘르세틴-3-O-자일로시드와 퀘르세틴-3-O-람노시드 1:1 (mole : mole) 복합물이 퀘르세틴-3-O-자일로시드만 단독으로 처리할 때보다 인슐린 저항성이 유도 된 근육세포에서 포도당 흡수를 더욱 촉진 시키는 상승효과를 가진다는 결과를 보여준다. Dulbecco's C2C12 muscle cells were dispensed into a 96-well black plate at a density of 0.5 × 104 cells/well and contained 1% penicillin streptomysin and 2% Horse serum (Gibco, USA). Change the medium with Modified Eagle's Medium High Glucose (DMEM HG medium; Hyclone, USA) to differentiate muscle cells for 7 days and add Dexamethasone (Sigma aldrich, USA) to Dulbecco's Modified Eagle's Medium Low Glucose (DMEM LG medium; Welgene, USA). Insulin-resistance was induced by treatment at a concentration of 1 μM for 16 hours. After induction, the culture medium was removed and starvation was performed in DPBS (Dulbecco's Phosphate-Buffered Saline, 1X) for 1 hour. After removing DPBS, make Dexamethasone (Sigma aldrich, USA) at a concentration of 1 μM in Dulbecco's Modified Eagle's Medium Glucose Free (DMEM GF medium; Welgene, USA) and make Quercetin-3-O-xyloside, Quercetin-3-O-rhamnoside Was treated at a concentration of 20 μM for 3 hours and 30 minutes. In addition, a 1:1 (mole: mole) Mixture of Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside at 10 μM concentration was prepared and treated for 3 hours and 30 minutes. After treatment with Quercetin-3-O-xyloside, Quercetin-3-O-rhamnoside, and Mixture, insulin (Sigma aldrich, USA) was treated at a concentration of 1 μM for 30 minutes. 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG: Invitrogen, USA) was treated at a concentration of 50 μM for 2 hours. After washing with phosphate buffered saline, the content of 2-NBDG in the cells was measured at 485 nm (exitation) and 535 nm (emission) with a fluorescence spectrophotometer (device manufacturer). The measured content of 2-NBDG is shown in Figure 6. As shown in FIG. 6, when each sample was treated at a concentration of 20 μM or 10 μM, when comparing the group treated with Insulin in Dexamethason and the group treated with Insulin in Dexamethason, and the control group treated with the sample, Quercetin-3-O-xyloside It was confirmed that the glucose uptake level of muscle cells treated with and Quercetin-3-O-rhamnoside, respectively, increased with the concentration. In addition, the glucose uptake level of the complex mixture of Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside at a concentration of 10 μM and 1:1 (mole: mole) was higher than when each single substance was treated. It could be confirmed that it increased. This is because quercetin-3-O-xyloside and quercetin-3-O-rhamnoside 1:1 (mole: mole) complexes induced insulin resistance more than when quercetin-3-O-xyloside was treated alone. The results show that it has a synergistic effect to further promote glucose absorption in muscle cells.
Claims (14)
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는, 당뇨병의 예방 또는 치료용 약학적 조성물. A pharmaceutical composition for preventing or treating diabetes, including quercetin-3-O-xyloside.
- 제1항에 있어서, 상기 약학적 조성물은 알파-글루코시다제 (α-Glucosidase)를 억제하는 활성을 가지는, 당뇨병의 예방 또는 치료용 약학적 조성물. According to claim 1, wherein the pharmaceutical composition has an activity to inhibit alpha-glucosidase (α-Glucosidase), a pharmaceutical composition for preventing or treating diabetes.
- 제1항에 있어서, 상기 약학적 조성물은 근육세포의 세포 내 포도당 흡수율을 증가시키는, 당뇨병의 예방 또는 치료용 약학적 조성물. The pharmaceutical composition for preventing or treating diabetes according to claim 1, wherein the pharmaceutical composition increases the intracellular glucose absorption rate of muscle cells.
- 제1항에 있어서, 상기 약학적 조성물은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 20 내지 40 μM 의 농도로 포함하는, 당뇨병의 예방 또는 치료용 약학적 조성물. According to claim 1, wherein the pharmaceutical composition is quercetin-3-O-xyloside (Quercetin-3-O-xyloside) containing a concentration of 20 to 40 μM, a pharmaceutical composition for preventing or treating diabetes.
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는, 당뇨병의 예방 또는 치료용 약학적 조성물. A pharmaceutical composition for preventing or treating diabetes, comprising quercetin-3-O-xyloside (Quercetin-3-O-xyloside) and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside).
- 제 5항에 있어서, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)의 몰비(mole ratio)는 1:10 내지 10:1인, 당뇨병의 예방 또는 치료용 약학적 조성물. The method of claim 5, wherein the mole ratio of quercetin-3-O-xyloside and quercetin-3-O-rhamnoside is 1:10 to 10:1, a pharmaceutical composition for preventing or treating diabetes.
- 제 5항에 있어서, 상기 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)를 0.1 내지 100 μM 의 농도로 포함하는, 당뇨병의 예방 또는 치료용 약학적 조성물. The method of claim 5, wherein the quercetin-3-O-xyloside and quercetin-3-O-rhamnoside (Quercetin-3-O-rhamnoside) at a concentration of 0.1 to 100 μM Containing as, a pharmaceutical composition for the prevention or treatment of diabetes.
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 포함하는, 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물. A health functional food composition for preventing or improving diabetes, including quercetin-3-O-xyloside.
- 제 8항에 있어서, 상기 건강기능성 식품 조성물은 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 20 내지 40 μM 의 농도로 포함하는, 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물. The health functional food composition of claim 8, wherein the health functional food composition comprises quercetin-3-O-xyloside in a concentration of 20 to 40 μM. .
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는, 당뇨병의 예방 또는 개선용 건강기능성 식품 조성물. Health functional food composition for preventing or improving diabetes, comprising quercetin-3-O-xyloside and quercetin-3-O-rhamnoside .
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)를 당뇨병 환자에게 투여하는 단계를 포함하는, 당뇨병의 예방 또는 치료방법. Quercetin-3-O- xyloside (Quercetin-3-O-xyloside) comprising the step of administering to a diabetic patient, a method of preventing or treating diabetes.
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 당뇨병 환자에게 투여하는 단계를 포함하는, 당뇨병의 예방 또는 치료방법. Prevention of diabetes, comprising administering quercetin-3-O-xyloside and quercetin-3-O-rhamnoside to a diabetic patient Or a method of treatment.
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도.Use of a pharmaceutical composition comprising quercetin-3-O-xyloside for preventing or treating diabetes.
- 퀘르세틴-3-O-자일로시드 (Quercetin-3-O-xyloside) 및 퀘르세틴-3-O-람노시드 (Quercetin-3-O-rhamnoside)을 포함하는 약학적 조성물의 당뇨병 예방 또는 치료 용도. Use of a pharmaceutical composition comprising quercetin-3-O-xyloside and quercetin-3-O-rhamnoside for preventing or treating diabetes.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20090091615A (en) * | 2008-02-25 | 2009-08-28 | 한국생명공학연구원 | Tetracera scandens extracts and 4h-chromen-4-one derivatives isolated therefrom increasing glucose uptake in differentiated l6 muscle cells |
| KR20100092786A (en) * | 2009-02-13 | 2010-08-23 | 대선주조 주식회사 | Multi-type antidiabetic food compound by lactobacillus fermented of natural bitterness compound from ganoderma lucidum, bitter melon and bitter buckwheat and preparing method of the same |
| KR20170043363A (en) * | 2015-10-13 | 2017-04-21 | 가톨릭대학교 산학협력단 | Composition comprising Quercetin-3-O-xyloside having immunostimulating activity |
-
2019
- 2019-10-11 KR KR1020190125961A patent/KR102261484B1/en active IP Right Grant
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2020
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20090091615A (en) * | 2008-02-25 | 2009-08-28 | 한국생명공학연구원 | Tetracera scandens extracts and 4h-chromen-4-one derivatives isolated therefrom increasing glucose uptake in differentiated l6 muscle cells |
| KR20100092786A (en) * | 2009-02-13 | 2010-08-23 | 대선주조 주식회사 | Multi-type antidiabetic food compound by lactobacillus fermented of natural bitterness compound from ganoderma lucidum, bitter melon and bitter buckwheat and preparing method of the same |
| KR20170043363A (en) * | 2015-10-13 | 2017-04-21 | 가톨릭대학교 산학협력단 | Composition comprising Quercetin-3-O-xyloside having immunostimulating activity |
Non-Patent Citations (4)
| Title |
|---|
| DAI XIAOQIAN, DING YE, ZHANG ZHAOFENG, CAI XIAXIA, LI YONG: "Quercetin and quercitrin protect against cytokine-induced injuries in RINm5F β-cells via the mitochondrial pathway and NF-κB signaling", INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, SPANDIDOS PUBLICATIONS, GR, vol. 31, no. 1, 1 January 2013 (2013-01-01), GR, pages 265 - 271, XP055799821, ISSN: 1107-3756, DOI: 10.3892/ijmm.2012.1177 * |
| KATHRYN L LIPSON, SONYA G FONSECA, FUMIHIKO URANO: "Endoplasmic Reticulum Stress-Induced Apoptosis and Autoimmunity in Diabetes.", CURRENT MOLECULAR MEDICINE., vol. 6, no. 1, 2006, pages 71 - 77, XP009527260, DOI: 10.2174/156652406775574613 * |
| LEE JISUN; CHOI JI WON; SOHNG JAE KYUNG; PANDEY RAMESH PRASAD; PARK YONG IL: "The immunostimulating activity of quercetin 3-O-xyloside in murine macrophages via activation of the ASK1/MAPK/NF-κB signaling pathway", INTERNATIONAL HNMUNOPHARMACOLOGY, ELSEVIER, AMSTERDAM, NL, vol. 31, 17 December 2015 (2015-12-17), NL, pages 88 - 97, XP029413017, ISSN: 1567-5769, DOI: 10.1016/j.intimp.2015.12.008 * |
| SEO JEONG YEON, PANDEY RAMESH PRASAD, LEE JISUN, SOHNG JAE KYUNG, NAMKUNG WAN, PARK YONG IL: "Quercetin 3-O-xyloside ameliorates acute pancreatitis in vitro via the reduction of ER stress and enhancement of apoptosis", PHYTOMEDICINE, ELSEVIER, AMSTERDAM, NL, vol. 55, 1 March 2019 (2019-03-01), AMSTERDAM, NL, pages 40 - 49, XP055799827, ISSN: 0944-7113, DOI: 10.1016/j.phymed.2018.07.011 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113440515A (en) * | 2021-08-20 | 2021-09-28 | 中国农业科学院郑州果树研究所 | Synergistic blood sugar reducing composition containing isoliquiritigenin and application thereof |
| CN113440515B (en) * | 2021-08-20 | 2022-08-12 | 中国农业科学院郑州果树研究所 | Synergistic blood sugar reducing composition containing isoliquiritigenin and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| KR102261484B1 (en) | 2021-06-04 |
| KR20210043152A (en) | 2021-04-21 |
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