Composition for improving liver health comprising gallocatechin gallate and isoquercitrin

Abstract

To provide a composition for improving liver health.SOLUTION: In one aspect, the composition of the present invention includes a complex component of Gallocatechin Gallate and Isoquercitrin, which effectively reduces oxidative stress and chronic inflammatory reaction that occur during the accumulation of lipids in liver tissue, thereby improving liver health and enabling the prevention, improvement, or treatment of various liver diseases such as hepatitis, cirrhosis, and liver cancer.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Disclosed herein are compositions for promoting liver health. [Background technology]

[0002] Hepatocytes play a key role in regulating the energy homeostasis of our body. While surplus energy ingested from food is stored in the form of macromolecular complexes, for example, glucose as glycogen and fatty acids as neutral lipids, when energy is insufficient due to hunger, glycogen and neutral lipids are broken down from the stored energy source and transported to tissues that require nutrients for use. However, when energy homeostasis is disrupted due to a high amount of nutritional intake compared to energy consumption, the excess energy is stored in fat cells, but when the storage limit of fat cells is exceeded, neutral fat accumulates in other tissues such as the liver and muscles in addition to fat. In particular, when fat accumulates in hepatocytes and the percentage of fat in a normal liver exceeds 5%, it is called fatty liver. Oxidative stress and chronic inflammatory responses that occur during the process of lipid accumulation in liver tissues can lead to various liver diseases such as hepatitis, cirrhosis, and liver cancer. Because the liver plays many important roles necessary for survival, a deterioration in liver health can change the metabolism of the entire body and become life-threatening. In particular, the liver is called the "silent organ," and even if the liver is damaged to a certain extent, it is difficult to notice or become aware of abnormal symptoms, which is also an obstacle to liver health management. Furthermore, in the case of fatty liver, since specialized therapeutic drugs targeting fatty liver have not yet been developed, it has been possible to manage it only by indirect treatment with drugs for lipid metabolism disorders such as hyperlipidemia (e.g., fibrates, statins, etc.), or by prevention through improvement of diet and lifestyle habits. Therefore, there is an urgent need to develop targeted therapeutic drugs and health functional foods that can improve liver health and prevent or treat fatty liver and liver diseases. [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] The present invention aims to provide a composition for promoting liver health. [Means for solving the problem]

[0005] To achieve the above object, in one aspect, the present invention provides a composition for promoting liver health, comprising 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 effectively reduces oxidative stress and chronic inflammatory responses that occur during lipid accumulation in liver tissue, thereby improving liver health and can be used to prevent, improve or treat various liver diseases such as hepatitis, cirrhosis, and liver cancer. [Brief description of the drawings]

[0007] [Figure 1] FIG. 1 is a graph comparing the improving effect of a mixture (Adiphenon) of gallocatechin gallate (GCG) and isoquercitrin (IQC) at different compounding ratios on non-alcoholic fatty liver. [Diagram 2] FIG. 1 is a graph comparing the improving effect of alcoholic fatty liver depending on the blend ratio of Adiphenon. [Diagram 3] FIG. 1 is a graph comparing the inhibitory effect of Adiphenon on the generation of reactive oxygen species in hepatocytes depending on the compounding ratio. [Figure 4]FIG. 13 is a graph comparing the inhibitory effect of different Adiphenon combination ratios on gene expression mediated by inflammatory response in hepatocytes. [Diagram 5] FIG. 13 is a graph comparing the inhibitory effect of different Adiphenon combination ratios on gene expression mediated by inflammatory response in hepatocytes. [Figure 6] FIG. 1 is a graph comparing the lipid accumulation-inhibitory effect in hepatocytes at specific blend ratios of APGQ.

[0008] In Figures 1 to 6, 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] The term "prevention" as used herein refers to any action that inhibits or delays liver disease by application of the pharmaceutical composition, and the term "treatment" refers to any action that improves or beneficially alters the condition of an individual susceptible to or suffering from liver disease by application of the pharmaceutical composition.

[0011] The term "ameliorating" as used herein refers to any action in which the liver disease is improved or beneficially altered by application of the composition.

[0012] 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.

[0013] In one aspect, the present invention relates to a composition for promoting liver health, which comprises gallocatechin gallate (GCG) and isoquercitrin (IQC) as active ingredients.

[0014] In an exemplary embodiment, the composition may be a pharmaceutical composition for preventing or treating liver disease.

[0015] In one exemplary implementation, the composition may be a food composition for preventing or improving liver disease.

[0016] 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.

[0017] In an exemplary embodiment, the gallocatechin gallate and isoquercitrin may be present 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, 1:2.3 or more, or 1:2.4 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, or 1:2.7 or less. When the gallocatechin gallate and isoquercitrin are contained in the above weight ratio, liver health can be effectively promoted. 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.

[0018] In one exemplary implementation, the liver disease may be at least one selected from the group consisting of hepatitis, fatty liver, cirrhosis, and liver cancer.

[0019] In one exemplary implementation, the liver disease may be alcoholic liver disease.

[0020] In one exemplary implementation, the composition may be applied to a subject in need of reducing oxidative stress or inflammation in liver cells.

[0021] In one exemplary embodiment, the composition may be applied to a subject in need of reducing expression of at least one gene selected from the group consisting of IL-1b, IL-6, and iNOS.

[0022] In one exemplary embodiment, the composition may be applied to a subject in need of inhibiting lipid accumulation in liver cells.

[0023] 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 dosage has a good effect on promoting liver health, and if it is less than the above dosage, the effect on promoting liver health is small, and if it is more than the above dosage, problems such as toxicity may occur. The above dosage may be divided into once a day or 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.

[0024] 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.

[0025] 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.

[0026] 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.

[0027] 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.

[0028] 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.

[0029] 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.

[0030] 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.

[0031] In another aspect, the present invention relates to a method for promoting liver health, comprising administering to a subject a composition comprising effective amounts of gallocatechin gallate (GCG) and isoquercitrin (IQC).

[0032] In still another aspect, the present invention relates to the use of gallocatechin gallate (GCG) and isoquercitrin (IQC) for the manufacture of a composition for promoting liver health.

[0033] In yet another aspect, the present invention relates to Gallocatechin Gallate (GCG) and Isoquercitrin (IQC) for promoting liver health.

[0034] In yet another aspect, the present invention relates to non-therapeutic uses of gallocatechin gallate (GCG) and isoquercitrin (IQC) to promote liver health.

[0035] 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.

[0036] [Example] [Example 1] Comparison of lipid accumulation suppression effects in hepatocytes by treatment with a mixture of gallocatechin gallate (GCG) and isoquercitrin (IQC) (AdiPhenon) Fatty liver is a condition in which excess neutral lipids accumulate in liver tissue (hepatocytes). Here, we investigated whether a mixture of gallocatechin gallate (GCG) and isoquercitrin (IQC) (AdiPhenon) can suppress lipid accumulation in hepatocytes. Specifically, HepG2 human hepatoma cell line purchased from ATCC was cultured in Roswell Park Memorial Institure-1640 medium (Sigma Aldrich) containing fetal bovine serum (Gibco) and 1% penicillin / streptomycin (P / S; Sigma Aldrich). To confirm the effect of GCG and IQC on improving fatty liver, HepG2 cells were pretreated with various ratios (GCG:IQC = 10:0 to 0:10) of the complex at a total concentration of 100 μg / mL for 2 hours, and then treated with oleic acid (500 μM; Sigma Aldrich; Figure 1) or ethanol (2 mM; Sigma Aldrich, Figure 2) for 72 hours to induce fatty liver. The cells were then washed twice with phosphate-buffered saline (PBS; Sigma Aldrich) and fixed with 10% formaldehyde (Sigma Aldrich) solution for 5 minutes, and the accumulation of neutral lipids in hepatocytes was confirmed with Nile Red (Sigma Aldrich; 300 nM) solution, which specifically stains neutral lipids. The amount of accumulated neutral lipids was quantified using a Tecan Infinite M200 Microplate Reader (ex 495 nm, em 585 nm).

[0037] As a result, as shown in Figures 1 and 2, when various combination ratios of AdiPhenon were administered, lipid accumulation in hepatocytes was significantly reduced regardless of the fatty liver induction factor, and the reduction effect was most excellent at a GCG:IQC ratio of 3:7 to 4:6.

[0038] [Example 2] AdiPhenon treatment alleviates oxidative stress and chronic inflammatory responses in hepatocytes Alcohol generates a large amount of reactive oxygen during the decomposition process, and excessive intake of fat increases the concentration of free fatty acids. The innate immune response that protects cells from external harmful substances mainly begins with the binding of TLR4 to lipopolysaccharides, and TLR4 secretes various inflammatory factors through signaling systems such as NF-κB and AP-1. However, since TLR4 recognizes not only endotoxins such as lipopolysaccharides but also free fatty acids, the presence of excessive free fatty acids induces undesirable immune responses (e.g., chronic inflammation) even in the absence of external harmful substances. Since prolonged immune responses are the main cause of secondary liver diseases such as cirrhosis, liver fibrosis, and liver cancer, reducing oxidative stress and inflammatory responses can help prevent and improve fatty liver and related liver diseases.

[0039] Therefore, additional experiments were performed to investigate whether AdiPhenon can improve oxidative stress and inflammatory responses induced during the development of fatty liver. Specifically, HepG2 cells were pretreated with various combination ratios of AdiPhenon at a concentration of 100 μg / mL for 2 hours, and then treated with oleic acid (500 μM) or ethanol (2 mM) for 24 hours. The same experimental set was prepared twice, and some were washed twice with PBS as in Example 1, and the cells were fixed with 10% formaldehyde. Then, the cells were washed two more times with PBS to remove residual formaldehyde, and treated with H2-DCFDA (20 μM, Invitrogen by Thermo Fisher Scientific) that can detect reactive oxygen for 30 minutes. The amount of reactive oxygen generated was observed using a Tecan Infinite M200 Pro Microplate Reade (ex 492 nm, em 517 nm). The remaining experimental set was used for mRNA expression analysis to observe the inflammatory response. RNA was extracted using the TaKaRa MiniBEST Universal RNA Extraction Kit (Takara Bio) and then quantified using the NanoQuant add-on device of the Tecan Infinite M200 (Tecan). The same amount (1 mg) of RNA was then quantified using RevertAid 1 stcDNA was synthesized using a one-strand cDNA Synthesis Kit (Thermo Fisher Scientific). To observe the expression of genes involved in inflammatory responses, the coding sequence of the target gene was used as a template and PCR was performed using NCBI primer BLAST ( https: / / www.ncbi.nlm.nih.gov / tools / primer-blast / ) service, primers were designed and prepared (Bioneer), and the expression of the relevant genes was observed using the synthesized cDNA and primers in a CFX96 thermocycler (Bio-Rad).

[0040] As a result, as shown in Figures 3, 4, and 5, the effects of various blend ratios of AdiPhenon on active oxygen and inflammatory responses were confirmed, and it was found that a ratio of 3:7 to 4:6 was the most effective.

[0041] [Example 3] Comparison of effects by AdiPhenon ratio There are various causes of fatty liver, such as alcoholic fatty liver caused by excessive drinking and non-alcoholic fatty liver caused by excessive intake of nutrients in the diet. Regardless of the cause, fatty liver is accompanied by oxidative stress and chronic inflammation during the onset process, so in the following examples, oleic acid, which causes excessive lipid accumulation in cells, was used. In order to find the optimal ratio that shows the best synergistic effect by the blending ratio of AdiPhenon, various treatments were performed at a ratio of GCG:IQC = 4:6 (about 1:1.5) to 2:8 (about 1:4), and then the experiment of Example 1 was repeated.

[0042] As a result, as shown in FIG. 6, the best lipid accumulation inhibitory effect was observed when the GCG:IQC ratio was between 1:2.4 and 1:2.7.

Claims

1. A composition for promoting liver health, comprising gallocatechin gallate (GCG) and isoquercitrin (IQC) as active ingredients.

2. The composition according to claim 1 , wherein the composition is a pharmaceutical composition for preventing or treating a liver disease.

3. The composition according to claim 1 , wherein the composition is a food composition for preventing or improving liver disease.

4. The composition according to claim 2 or 3, wherein the liver disease is at least one selected from the group consisting of hepatitis, fatty liver, cirrhosis, and liver cancer.

5. The composition according to claim 2 or 3, wherein the liver disease is alcoholic liver disease.

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

9.

7. The composition of claim 1, wherein the composition is applied to a subject in need of reducing oxidative stress or inflammation in liver cells.

8. The composition according to claim 7, wherein the composition is applied to a subject in need of reducing expression of at least one gene selected from the group consisting of IL-1b, IL-6, and iNOS.

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

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