Composition for controlling blood sugar comprising gallocatechin gallate and isoquercitrin

Abstract

To provide a composition having excellent blood sugar control capability.SOLUTION: In an aspect, a composition of the present invention including a complex component of Gallocatechin Gallate and Isoquercitrin can effectively inhibit gluconeogenesis in liver cells and effectively improve glucose transport in differentiated fat cells and liver cells, thereby exhibiting excellent blood sugar control efficacy.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Disclosed herein is a composition having excellent glycemic control properties. [Background technology]

[0002] Glucose is absorbed into the body through the processes of food intake, digestion, and absorption, and may also be newly produced by gluconeogenesis in liver cells. On the other hand, glucose is used as an energy source in cells with active energy metabolism, such as nerve cells and muscle cells, or it reduces blood glucose concentration by promoting the transport of glucose into cells for the purpose of storing surplus energy in fat cells, etc. In this way, blood glucose control is balanced through various processes, but if blood glucose control fails due to various causes (for example, fasting blood glucose level is 126 mg / dl or higher), diabetes develops. Diabetes not only causes various metabolic diseases such as obesity, hyperlipidemia, and hypertension, but is difficult to cure once it develops, and requires lifelong management of diet and lifestyle, so it is a disease that greatly reduces the quality of life. Therefore, there is a demand for the development of health functional foods or pharmaceuticals that can prevent or improve complications of metabolic diseases and improve the quality of life by suppressing the onset of diabetes or improving the symptoms of diabetes. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Korean Patent Publication No. 2022-0145604 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a composition having excellent blood sugar control ability. [Means for solving the problem]

[0005] In order to achieve the above-mentioned object, in one aspect, the present invention provides a composition for blood sugar control, which comprises gallocatechin gallate (GCG) and isoquercitrin (IQC) as active ingredients. Effect of the Invention

[0006] In one aspect, the composition of the present invention contains a complex component of gallocatechin gallate and isoquercitrin, which can effectively inhibit gluconeogenesis in hepatocytes and effectively promote glucose transport in differentiated adipocytes and hepatocytes, thereby exhibiting excellent blood glucose control efficacy. [Brief description of the drawings]

[0007] [Figure 1] FIG. 1 is a graph comparing the inhibitory effect of a mixture (AdiPhenon) of gallocatechin gallate (GCG) and isoquercitrin (IQC) on the expression of gluconeogenesis-related genes in hepatocytes, depending on the compounding ratio. [Diagram 2] FIG. 1 is a graph comparing the glucose absorption promoting effect in adipocytes depending on the blending ratio of Adiphenon. [Diagram 3] FIG. 1 is a graph comparing the glucose transport promoting effect in muscle cells depending on the mixing ratio of Adiphenon. [Figure 4] FIG. 13 is a graph comparing changes in the expression of citric acid cycle-related genes in adipocytes depending on the mixing ratio of Adiphenon. [Diagram 5] FIG. 1 is a graph comparing changes in the expression of citric acid cycle-related genes in muscle cells depending on the combination ratio of Adiphenon. [Figure 6] FIG. 1 is a graph comparing changes in lactate accumulation in cells due to Adiphenon treatment. [Figure 7] FIG. 1 is a graph comparing the effect of suppressing the expression of gluconeogenesis genes at specific combination ratios of Adiphenon. [Figure 8]FIG. 1 is a graph comparing the glucose absorption promoting effect in adipocytes according to the specific compounding ratio of Adiphenon. [Figure 9] FIG. 1 is a graph comparing the glucose absorption promoting effect in muscle cells according to the specific combination ratio of Adiphenon.

[0008] In Figures 1 to 9, experimental groups showing statistically significant differences (P<0.05) are indicated with different symbols. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present invention will be described in detail below.

[0010] In one aspect, the present invention relates to a composition for controlling blood sugar, comprising gallocatechin gallate (GCG) and isoquercitrin (IQC).

[0011] In an exemplary embodiment, the gallocatechin gallate and isoquercitrin may be contained in a weight ratio of 9:1 or more, 8:2 or more, 7:3 or more, 6:4 or more, 5:5 or more, or 4:6 or more, or may be contained in a weight ratio of 1:9 or less, or 2:8 or less. For example, the gallocatechin gallate and isoquercitrin may be contained in a weight ratio of 9:1 to 1:9, but is not limited thereto.

[0012] In an exemplary embodiment, the gallocatechin gallate and isoquercitrin may be included in a weight ratio of 1:1 or more, 1:1.1 or more, 1:1.2 or more, 1:1.3 or more, 1:1.4 or more, 1:1.5 or more, 1:1.6 or more, 1:1.7 or more, 1:1.8 or more, 1:1.9 or more, 1:2.0 or more, 1:2.1 or more, 1:2.2 or more, or 1:2.3 or more, or in a weight ratio of 1:9 or less, 1:8 or less, 1:7 or less, 1:6 or less, 1:5 or less, 1:4 or less, 1:3.5 or less, 1:3.4 or less, 1:3.3 or less, 1:3.2 or less, 1:3.1 or less, 1:3 or less, 1:2.9 or less, 1:2.8 or less, 1:2.7 or less, 1:2.6 or less, or 1:2.5 or less. When the gallocatechin gallate and isoquercitrin are contained in the above weight ratio, blood glucose level can be effectively controlled. For example, the gallocatechin gallate and isoquercitrin may be contained in a weight ratio of 1:1 to 1:9 or 1:1.5 to 1:4, but is not limited thereto.

[0013] The term "application" as used herein means providing the composition to a subject by any suitable method, including administration, spreading, absorbing, and ingestion, etc. In this case, the subject means any animal, such as a human, monkey, dog, goat, pig, or rat, to which the composition can be applied.

[0014] In one exemplary implementation, the glycemic control may be hypoglycemic. The active ingredient of the composition may inhibit glycolysis in the blood or promote cellular absorption of glucose in the blood.

[0015] Blood glucose concentration is regulated by the balance between glucose inflow / production and consumption / storage. When glucose concentration is low during fasting, pancreatic α cells detect hypoglycemia and secrete glucagon to break down glycogen stored in liver cells or synthesize glucose from acetyl coenzyme A. This process is called gluconeogenesis, which is suppressed by insulin secreted during full stomach. If the insulin signaling system is impaired for some reason and insulin cannot perform its role properly despite its presence (insulin resistance), liver cells will continue to produce glucose unless insulin inhibits gluconeogenesis, which may lead to hyperglycemia.

[0016] In one exemplary embodiment, the composition may be administered to a subject in need of reducing gluconeogenesis in liver cells.

[0017] In one exemplary embodiment, the composition may be characterized in that it is applied to a subject in need of reducing expression of a gene involved in gluconeogenesis, and the gene involved in gluconeogenesis may be a PEPCK (Phosphoenol Pyruvate CarboxyKinase) or G6Pase (Glucose 6-Phosphatase) gene.

[0018] In one exemplary implementation, the composition may be applied to a subject in need of increased glucose uptake and / or transport in fat or liver cells.

[0019] In an exemplary embodiment, the composition may be applied to a subject in need of increasing expression of a citric acid cycle component gene, and the citric acid cycle component gene may be isocitrate dehydrogenase (IDH) or malate dehydrogenase (MD).

[0020] In one exemplary implementation, the composition may be applied to a subject in need of inhibiting lactate accumulation in cells.

[0021] In one exemplary implementation, the composition may be applied to a subject in need of promoting aerobic energy metabolism in cells.

[0022] The term "prevention" as used herein refers to any action of suppressing or delaying diabetes, diabetic complications, or glucose metabolic disorders by application of the pharmaceutical composition. The term "treatment" refers to any action of improving or beneficially altering the symptoms of an individual suspected of or suffering from diabetes, diabetic complications, or glucose metabolic disorders by application of the pharmaceutical composition.

[0023] The term "amelioration" as used herein refers to any action in which diabetes, diabetic complications or glucose metabolic disorders are improved or beneficially altered by application of the composition.

[0024] In an exemplary embodiment, the composition may be a pharmaceutical composition for preventing or treating diabetes, diabetic complications, or glycometabolic disorders.

[0025] In one exemplary embodiment, the composition may be a food composition for preventing or improving diabetes, diabetic complications, or glucose metabolism disorders.

[0026] In this specification, the term "diabetes" refers to a disease that occurs when the balance between insulin sensitivity and insulin secretion in the body is lost, that is, when there is insulin deficiency or insulin sensitivity is reduced, blood glucose is not utilized, blood glucose is not controlled, and glucose is excreted in urine. The diabetic complications are broadly classified into acute complications and chronic complications. Examples of the acute complications include ketoacidosis, hyperosmolar hyperglycemic syndrome, hypoglycemia, etc., and examples of the chronic complications include macrovascular complications such as coronary artery disease, cerebrovascular disease, and peripheral vascular disease, small vascular complications such as diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy, and diabetic foot disease, but are not limited thereto.

[0027] In an exemplary embodiment, the daily dosage of the active ingredient of the composition may be 5 mg / kg or more, 6 mg / kg or more, 7 mg / kg or more, 8 mg / kg or more, 9 mg / kg or more, 10 mg / kg or more, 20 mg / kg or more, 30 mg / kg or more, 40 mg / kg or more, 50 mg / kg or more, 60 mg / kg or more, 70 mg / kg or more, 80 mg / kg or more, 90 mg / kg or more, 100 mg / kg or more, 1 g / kg or more, 2 g / kg or more, 3 g / kg or more, 4 g / kg or more, 5 g / kg or more, or 6 g / kg or more, or 60 g / kg or less, 59 g / kg or less, 58 g / kg or less, 57 g / kg or less, 56 g / kg or less, 55 g / kg or less, 54 g / kg or less, 53 g / kg or less, 52 g / kg or less, or 51 g / kg or less. The above-mentioned dosage has excellent blood glucose control effect, and if it is less than the above-mentioned dosage, the blood glucose control effect is small, and if it is more than the above-mentioned dosage, problems such as toxicity may occur. The above-mentioned dosage may be applied once a day or divided into several times. For example, it may be applied 2 to 24 times a day, 1 to 2 times every 3 days, 1 to 6 times a week, 1 to 10 times a week, 1 to 15 times a week, 1 to 3 times a week, or 1 to 12 times a year, but is not limited thereto.

[0028] In an exemplary embodiment, the pharmaceutical composition may be provided in any dosage form suitable for local application. For example, it may be administered orally, transdermally, intravenously, intramuscularly, or subcutaneously. For example, the pharmaceutical composition may be, but is not limited to, an injection, a solution for external use on the skin, a suspension, an emulsion, a gel, a patch, or a spray. The dosage form may be easily prepared according to a conventional method in the art, and may appropriately use a surfactant, an excipient, a hydrating agent, an emulsifying agent, a suspending agent, a salt or a buffer for adjusting osmotic pressure, a coloring agent, a spice, a stabilizer, an antiseptic, a preservative, or other commonly used auxiliary agent.

[0029] In one exemplary implementation, the active ingredient of the pharmaceutical composition will depend on the subject's age, sex, weight, medical condition and its severity, the route of administration, or the prescriber's judgment. Determination of the appropriate dosage based on such factors is within the level of one of ordinary skill in the art.

[0030] In an exemplary embodiment, when the composition is used as an additive for health functional food, it can be added as it is, or can be used in combination with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment. The dosage form of the health functional food can be any of powder, granule, pill, tablet, capsule, and general food or drink forms.

[0031] In an exemplary embodiment, the type of the health functional food is not particularly limited, and examples of foods to which the composition can be added include meat, confectionery, noodles, gums, dairy products such as ice cream, various soups, soft drinks, tea, energy drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the usual sense.

[0032] In one exemplary embodiment, the composition may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on 100 parts by weight of raw materials, during the preparation of the health functional food or beverage. However, for long-term intake for health and hygiene purposes or health regulation purposes, the amount may be less than the above range.

[0033] In an exemplary embodiment, the beverage in the health functional food may contain various flavorings, natural carbohydrates, etc. as additional ingredients, like ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrates may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the beverage according to the present invention, but is not limited thereto.

[0034] In an exemplary embodiment, in addition to the above, the health functional food according to the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonation agents used in carbonated drinks. In addition, the health functional food according to the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used alone or in combination. The proportion of these additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food according to the present invention.

[0035] In another aspect, the present invention relates to a method for controlling blood sugar, comprising administering to a subject a composition comprising effective amounts of gallocatechin gallate (GCG) and isoquercitrin (IQC).

[0036] In still another aspect, the present invention relates to the use of gallocatechin gallate (GCG) and isoquercitrin (IQC) for producing a composition for blood glucose control.

[0037] In still another aspect, the present invention relates to gallocatechin gallate (GCG) and isoquercitrin (IQC) for blood sugar control.

[0038] In another aspect, the present invention relates to non-therapeutic uses of gallocatechin gallate (GCG) and isoquercitrin (IQC) for glycemic control.

[0039] The configuration and effects of the present invention will be described in more detail below with reference to examples. However, the following examples are provided to aid in understanding the present invention, and are not intended to limit the scope and spirit of the present invention.

[0040] [Example] [Example 1] Comparison of gluconeogenesis inhibitory effects in hepatocytes treated with a mixture of gallocatechin gallate (GCG) and isoquercitrin (IQC) (AdiPhenon) The present inventors confirmed whether a mixture of gallocatechin gallate (GCG) and isoquercitrin (IQC) (AdiPhenon) can inhibit gluconeogenesis in hepatocytes. Specifically, HepG2 cells purchased from ATCC were cultured in Roswell Park Memorial Institure-1640 medium (Sigma Aldrich) containing 10% fetal bovine serum (Gibco) and 1% penicillin / streptomycin (P / S; Sigma Aldrich). In order to confirm the gluconeogenesis inhibitory effect of the GCG and IQC ratio, HepG2 cells were treated with AdiPhenon complexes at various ratios (GCG:IQC = 10:0 to 0:10) at a total concentration of 100 μg / mL for 24 hours. As a positive control, insulin (10 nM, Sigma Aldrich), which is known to inhibit gluconeogenesis, was used. The cells were then washed with phosphate-buffered saline (PBS; Sigma Aldrich) and RNA was extracted using the TaKaRa MiniBEST Universal RNA Extraction Kit (Takara Bio). The same amount (1 mg) of RNA was extracted with RevertAid 1 st cDNA was synthesized using a -strand cDNA Synthesis Kit (Thermo Fisher Scientific), and the expression of the rate-limiting enzymes of gluconeogenesis, PEPCK (phosphoenol pyruvate carboxylkinase) and G6Pase (glucose 6-phosphatase), was observed using a CFX96 thermocycler (Bio-Rad) (Figure 1).

[0041] As a result, as shown in Figure 1, Adiphenon suppressed the expression of gluconeogenesis-related genes in the single substance treatment group regardless of the compounding ratio, and the suppression of the expression of the above genes by Adiphenon was most excellent at the ratio of GCG:IQC = 4:6 to 2:8. In other words, it was confirmed that Adiphenon can effectively inhibit gluconeogenesis in hepatocytes.

[0042] [Example 2] Stimulation of glucose transport in adipocytes and muscle cells by AdiPhenon treatment To confirm another major mechanism of glycemic control, we examined whether Adiphenon could promote glucose uptake in adipocytes and muscle cells. Specifically, mouse preadipocytes (3T3-L1) and myoblasts (C2C12) were purchased from ATCC. 3T3-L1 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM; Sigma Aldrich) containing 10% bovine calf serum (Gibco) and 1% P / S, and C2C12 cells were cultured in DMEM containing 10% FBS and 1% P / S. Adipocyte differentiation was induced as follows. 3T3-L1 cells were grown to 100% fullness in a culture dish and then treated with DMEM containing dexamethasone (1 mM; Sigma Aldrich), insulin (10 mg / mL; Sigma Aldrich), 3-isobutyl-1-methylxanthine (0.5 mM; Sigma Aldrich), FBS (10%) and P / S (1%) for 48 h. The cells were then cultured with DMEM containing 10% FBS, 10 mg / mL insulin and 1% P / S every 2 days for 14 days to induce complete adipocyte differentiation. For myocyte differentiation, C2C12 cells were grown to 100% fullness in a culture dish and then differentiated for 7 days in DMEM medium containing 2% horse serum (HS; Gibco) and 1% P / S. The medium was changed every day during the differentiation process. To confirm glucose transport in differentiated adipocytes and hepatocytes, starvation was induced using serum-free, low-glucose DMEM (Sigma Aldrich) containing only 1% P / S on the day before the experiment. Thereafter, similar to Example 1, various ratios of Adiphenon were treated for 24 hours, and an experiment group was further generated in which the cells were treated with insulin (100 nM) to confirm the interaction with insulin. After 24 hours, the amount of glucose transported into the cells was quantified using a Glucose Uptake Assay kit (colorimetric, abcam). Quantification was performed using a Tecan Infinite M200 Pro Microplate Reader (ab 412 nm).

[0043] As a result, as shown in Figures 2 and 3, it was confirmed that Adiphenon at various compounding ratios can effectively promote intracellular glucose absorption in fat cells and muscle cells, and such effect is doubled in the presence of insulin. It was also confirmed that the glucose transport improving effect of Adiphenon is most excellent at a GCG:IQC ratio of 4:6 to 2:8.

[0044] [Example 3] Activation of the citric acid cycle in cells treated with AdiPhenon Glucose that enters cells is either used as an energy source or stored by cells in the form of glycogen, fatty acids, etc. In particular, if it is stored in fat cells as fatty acids or neutral lipids, it can cause obesity due to increased body fat, so it is necessary to check whether the absorbed glucose is being used as an energy source. Through this process, glucose is broken down into two molecules of pyruvate, which is then converted into acetyl coenzyme A through further metabolism. Acetyl coenzyme A is then used for ATP synthesis in the mitochondrial electron transport system via the citric acid cycle.

[0045] Therefore, to examine whether Adiphenon treatment can activate intracellular energy metabolism, fat cells and muscle cells were differentiated by the method described in Example 2, and then treated with Adiphenon at various mixing ratios for 24 hours. After that, RNA was extracted and cDNA was synthesized by the method described in Example 1, and the expression of citric acid cycle component genes was confirmed.

[0046] As a result, as shown in Figures 4 and 5, it was confirmed that the expression of citric acid cycle component genes was promoted by treatment with Adiphenon at various ratios in both fat cells (Figure 4) and muscle cells (Figure 5). It was also confirmed that the effect of Adiphenon was most excellent when the GCG:IQC ratio was 3:7 to 2:8.

[0047] On the other hand, even if the expression of the citric acid cycle component genes increases, the citric acid cycle cannot operate smoothly if there is a shortage of oxygen or mitochondria in the cells. In such a situation, instead of producing acetyl coenzyme A, pyruvate produces a small amount of ATP through the lactic acid fermentation process, and lactic acid accumulates during this process, causing intracellular fatigue. Therefore, by comparing the degree of lactic acid accumulation in the cells, the aerobic energy metabolism activity in the cells can be indirectly confirmed. When cells are treated with CoCl2, it induces the stabilization of HIF-1α, and the cells recognize that they are in a hypoxic state. At that time, due to the lack of oxygen (or the illusion that it is), the cells do not perform oxidative phosphorylation using mitochondria, and instead generate ATP using lactic acid fermentation, which does not consume oxygen.

[0048] Therefore, differentiated fat cells and muscle cells were treated with CoCl2 (100 μM; Sigma Aldrich) for 12 hours to induce hypoxia, and then treated with Adiphenone at GCG:IQC ratios of 3:7 and 2:8, which showed the most synergistic effect in the field of energy metabolism, for 24 hours, and the amount of lactate accumulated in the cells was measured (L-lactate Assay Kit (Colorimetric), Abcam).

[0049] As a result, as shown in FIG. 6, the experimental results that Adiphenon suppresses lactate accumulation in cells together with promoting the expression of citric acid cycle genes indicate that Adiphenon can promote aerobic energy metabolism in cells.

[0050] [Example 4] Comparison of effects by AdiPhenon ratio In order to find the optimal ratio of Adiphenone that would provide the best synergistic effect, the experiments in Examples 1 and 2 were repeated after treatment with various combination ratios within the range of GCG:IQC = 4:6 (approximately 1:1.5) to 2:8 (approximately 1:4).

[0051] As a result, as shown in Figures 7 to 9, it was confirmed that the most excellent synergistic effect was observed when the GCG:IQC ratio was between 1:2.1 and 1:2.7, more specifically, 1:2.4.

Claims

1. A composition for controlling blood sugar, comprising gallocatechin gallate (GCG) and isoquercitrin (IQC) as active ingredients.

2. The composition according to claim 1, comprising the gallocatechin gallate (GCG) and isoquercitrin (IQC) in a weight ratio of 9:1 to 1:

9.

3. The composition according to claim 1, characterized in that the composition is applied to a subject in need of reducing gluconeogenesis in hepatocytes.

4. The composition of claim 1, wherein the composition is applied to a subject in need of reducing expression of a gene involved in gluconeogenesis.

5. The composition of claim 1, wherein the composition is applied to a subject in need of increasing glucose absorption and / or transport in fat cells or liver cells.

6. The composition according to claim 1, wherein the composition is applied to a subject in need of increasing expression of a citric acid cycle component gene.

7. The composition according to claim 1, wherein the composition is applied to a subject in need of suppressing lactate accumulation in cells.

8. The composition according to claim 1, wherein the composition is applied to a subject in need of promoting aerobic energy metabolism in cells.

9. The composition according to claim 1 , which is a pharmaceutical composition for preventing or treating diabetes, diabetic complications or glucose metabolic disorders.

10. The composition according to claim 1 , which is a food composition for preventing or ameliorating diabetes, diabetic complications or glucose metabolic disorders.

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