KR20150091771A - Composition for treating or preventing diabetes contaning extract of caulerpa lentillifera - Google Patents

Abstract

The present invention relates to an anti-diabetes composition and, specifically, to a medical composition for treating or preventing diabetes or a food composition for preventing or alleviating diabetes. More specifically, the present invention relates to a medical composition for treating or preventing diabetes and a food composition for preventing or alleviating diabetes containing a Caulerpa Lentillifera extract as an active ingredient. As a result of confirmation using a result of an experiment related to extracellular enzymes and a result of an experiment using beta cells, muscular cells, and fat cells of the pancreas, it has been confirmed that the Caulerpa Lentillifera extract which is an active ingredient of the anti-diabetes composition has remarkably excellent anti-diabetes activities. A pharmaceutical composition for treating or preventing diabetes and a food composition for preventing or alleviating diabetes can be widely used in treating, preventing, or alleviating diabetes or diabetic complications.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a composition for treatment or prevention of diabetes, which comprises sea grape extract,

The present invention relates to a composition for the treatment, improvement or prevention of diabetes, comprising a composition for the treatment, improvement or prevention of diabetes, specifically, a composition derived from a natural substance as an active ingredient.

Diabetes mellitus refers to a chronic metabolic disorder in which insulin is insufficient for the intracellular uptake of glucose, or insulin acts abnormally and glucose can not be maintained at an appropriate level in the blood.

There are two types of diabetes: type 1 diabetes that does not cause insulin secretion in the pancreatic beta cells due to genetic factors and environmental factors, and type 2 diabetes that develops insulin resistance in peripheral tissues.

Specifically, the diabetes is divided into type 1 diabetes (insulin-dependent diabetes mellitus) and type 2 diabetes (non-insulin dependent diabetes mellitus) depending on the secretion and action of insulin. The type 1 diabetes is caused by deficiency of insulin which is produced in pancreatic beta cells and reduces the concentration of glucose in the blood, and it is necessary to supply insulin regularly from the outside. On the other hand, the type 2 diabetes mellitus has a relatively high ability to secrete insulin from the beta cells of the pancreas, but the secreted insulin does not play a role in the smooth functioning of the insulin. Thus, It is a combination of therapy. Most of the therapeutic agents for type 2 diabetes mellitus are hypoglycemic agents for oral administration.

The type 1 diabetes is the root cause of pancreatic beta cell death or damage, and oxidative stress is known as one of the causes of pancreatic beta cell damage. In addition, progressive pancreatic beta cell dysfunction due to oxidative stress further increases insulin resistance in type 2 diabetes.

These oxidative stresses are known to occur as a result of a large amount of reactive oxygen species (ROS) in the body or a decrease in the function of the antioxidant system, resulting in an imbalance between the oxidative system and the antioxidant system. When hyperglycemia persists, free radicals (H ₂ O ₂ , O ₂ ^- , HO ^- )) are generated to induce oxidative stress, resulting in damage to pancreatic beta cells, resulting in increased diabetes, diabetic retinopathy, diabetic nephropathy, Diabetic complications such as hypercholesterolemia, hypercholesterolemia, hypertriglyceridemia, and the like.

The most important goal in the treatment of diabetes is to regulate blood sugar levels as close to normal as possible. Therapeutic methods include pharmacotherapy, diet and exercise therapy.

Currently, oral hypoglycemic agents used in patients with type 1 and type 2 diabetes include α-glucosidase inhibitors, sulfonylurea agents, and biguanide agents. Specifically, metformin, a kind of the above-mentioned biguanide, thiazolidinedione-based pioglitazone and rosiglitazone, which activates PPARγ, α-glucosidase inhibitors that inhibit the absorption of carbohydrates in the small intestine to prevent rapid increase in postprandial blood glucose level, And sulphonylurea, which promotes insulin secretion in? -Cells. In recent years, a DPP-IV inhibitor has been developed that selectively inhibits DPP-IV enzyme, which directly degrades the incretin enzyme, which promotes secretion of insulin and lowers blood glucose, and maintains the incretin enzyme at high concentration for a long time And is commercially available.

  More specifically, the? -Glucosidase inhibitor shows the therapeutic effect of diabetes by decreasing postprandial blood glucose and elevation of blood insulin by delaying digestion and absorption of carbohydrates in the ingested diets. The α-glucosidase inhibitor promotes secretion in the small intestine of glucagon-like peptide-1, which does not induce hyperinsulinemia or hypoglycemia, promotes insulin secretion and inhibits glucagon secretion It has advantages.

Currently, clinical use includes acarbose, voglibose, and miglitol. In addition, tolbutamide, one of the sulfonylureas that stimulate pancreatic tissue to increase insulin secretion, may be used as an adjunct to insulin therapy.

However, in the case of long-term use, the therapeutic agent for the treatment of diabetes such as α-glycosidase inhibitor currently marketed and used may exhibit side effects such as abdominal bloating and vomiting diarrhea in some patients, and its use may be restricted.

To date, there has been no established cure for diabetes mellitus. Due to the adverse effects of drug treatment, there are many limitations on the use of drugs. Research on diabetes mellitus drugs from natural products has been actively conducted. WHO also has been working on diabetes mellitus , And efforts are underway to find active ingredients from natural plant-derived substances that are effective for diabetes.

KR10-1252149B KR10-2008-0114454A KR10-1148978B

The present invention provides a pharmaceutical composition for treating or preventing diabetes and a food composition for improving or preventing diabetes, which effectively treats or prevents diabetes, inhibits diabetes, and contains an extract of a natural plant harmless to the human body as an active ingredient .

In order to accomplish the above object, the present invention provides a pharmaceutical composition for treating or preventing diabetes comprising an extract of sea bream according to one embodiment as an active ingredient.

The sea grape extract may be one extracted with any one selected from the group consisting of alcohols having 1 to 5 carbon atoms, water, and mixtures thereof, preferably extracted with ethanol or an aqueous solution of ethanol.

The present invention also provides a food composition for improving or preventing diabetes, which comprises sea grape extract as an active ingredient.

The sea grape extract may be one selected from the group consisting of alcohols having 1 to 5 carbon atoms, water, and mixtures thereof.

The food composition may be a health functional food.

Unless otherwise specified in the present specification, an effective component means a component that provides an anti-obesity effect in a composition, specifically, a pharmaceutical composition or a food composition.

As used herein, unless otherwise specified, pharmacologically acceptable means physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like, when administered to an animal, preferably a human, do.

The inventors of the present invention have found that when a substance having a diabetic effect is studied from a natural plant-derived substance having secured human safety, the sea grape extract has an alpha-glucosidase inhibitory activity, a DPP-IV enzyme inhibitory activity and a PTP1B The inhibitory activity is remarkably superior to that of the oxinda extract, which is an inbred plant, and the glucose uptake effect is excellent in muscle cells and adipocytes, and the protective effect of pancreatic beta cells is remarkably higher than that of oxinda extract It is excellent and has an excellent effect of improving insulin secretion ability of pancreatic beta cells and has an effect on the expression of proteins related to insulin secretion, diabetes induction, insulin resistance or diabetic complication and has an excellent antidiabetic effect. The present invention has been accomplished on the basis of this finding.

Hereinafter, the present invention will be described in more detail.

The present invention sea grapes (Caulerpa lentillifera ) extract as an active ingredient. The composition for treating or preventing diabetes may be a pharmaceutical composition for treating or preventing diabetes.

The sea grape ( Caulerpa lentillifera ) is a seaweed in Okinawa, the Philippines, and Vietnam, and has been mainly used for food in Okinawa. The sea grape is one of the five seaweed delicacies that the Okinawan people eat, and is also famous as a longevity food for the residents of Okinawa Island. The sea grape is an alkaline food. It is rich in various minerals, vitamins and alginic acid as a dietary fiber. It is known to have excellent antioxidative activity, active oxygen and lipid peroxidation inhibiting ability, and helps the waste excretion and defecation.

As described above, the sea grape ethanol extract is a plant belonging to the same genus, Caulerpa it was confirmed that the inhibitory activity of the enzyme related to diabetes mellitus, the ability to protect pancreatic cells, the ability to promote insulin secretion, and the insulin sensitivity improving effect were remarkably superior to those of okamurai extract.

The sea grape extract may be one extracted by a method of extracting plants, specifically, a method of extracting seaweed, and may be, for example, an extract prepared by extracting with an extraction solvent, or a fraction prepared by fractionating the extract with a fraction solvent. The plant to be extracted from the sea grape extract may be the entire plant or a part thereof.

The extraction solvent may be at least one selected from the group consisting of water and an organic solvent. The organic solvent may be a polar solvent such as methanol, ethanol or the like, an alcohol having 1 to 5 carbon atoms, ethyl acetate or acetone, a nonpolar solvent such as benzene, hexane or dichloromethane, or a mixed solvent thereof, preferably 1 to 5 carbon atoms Of an alcohol, water and a mixture thereof, more preferably ethanol or an aqueous solution of 50 to 99% of ethanolol.

The sea grape extract of the present invention may be one prepared by a conventional plant extract, specifically, a method for producing seaweed extract. More specifically, extraction can be performed by adding an extraction solvent to sea grapes from which impurities have been removed. The extraction process may be a cold extraction method, a warm extraction method, a pressure extraction method, or an ultrasonic pulverization extraction method.

The fraction solvent may be water, butanol, ethyl acetate, chloroform, hexane or a mixture thereof. The fraction may be an extract prepared by the above extraction method, specifically, a fraction obtained by further fractionating the crude extract. The fraction solvent may be a solvent selected from the group consisting of ethyl acetate, ether, chloroform, benzene, hexane, methylene chloride and a mixed solvent thereof, preferably hexane. Specifically, the fractionation step is performed by sequentially adding hexane, chloroform, ethyl acetate, butanol, and water to the crude extract in this order, then sequentially fractionating the hexane fraction, the chloroform fraction, the ethyl acetate fraction, the butanol fraction, . ≪ / RTI >

After the extraction or fractionation, the extract or fraction may be subjected to a vacuum filtration process or may be further concentrated and / or lyophilized to concentrate or remove the solvent. The obtained extract can be stored in a deep freezer until use.

Specifically, the sea grape extract may be further subjected to a filtration step, a concentration step and / or a drying step after the solvent extraction step, or may be carried out by selectively or redundantly performing the filtration step. The filtration step may be performed using a filter paper, The concentration step can be carried out through reduced pressure concentration or the like, and the drying step can be performed by a method such as hot air drying or freeze drying.

In the present invention, diabetes is a disease in which the balance between insulin sensitivity and insulin secretion in the body is broken, that is, insulin deficiency or insulin sensitivity is lowered, . In addition, the diabetes may include diabetic complications.

The diabetes mellitus is classified into type 1 diabetes and type 2 diabetes. The diabetes is preferably caused by T-cell, B-cell, and macrophage, And may be type 1 diabetes known to cause the destruction and dysfunction of pancreatic beta cells.

In addition, the diabetic complication may be a cholesterol metabolism disorder-related disease such as hypercholesteremia, hypertriglyceridemia or atherosclerosis, thrombosis, atherosclerosis, microangiopathy, kidney disease , Heart disease, diabetic retinopathy or cataracts, and the like.

The cholesterol metabolism disorder-related disease refers to a disease caused by accumulation of cholesterol at an appropriate level or higher in the body or blood vessel due to abnormality in cholesterol regulation function or a disease caused thereby, and examples thereof include hyperlipidemia, hypercholesterolemia, hypertriglyceridemia A cholesterol metabolism disorder related disease such as atherosclerosis or atherosclerosis. The term " hyperlipidemia " means a disease caused by a high fat content in the blood, which can cause disease or a high content of blood fat.

The sea grape extract of the present invention is an in vitro test ( in vitro and cell experiments, it was found to have an excellent antidiabetic effect. As an example, unlike oocyte, which is an endophyte, after inducing apoptosis by treatment with IL-1β 2 ng / ml and IFN-γ 100 Units / ml in RINm5F, a beta cell line obtained from mouse pancreas, Beta cell viability and insulin production ability, it was confirmed that the sea grape extract had remarkably excellent antidiabetic effect as compared with oxinda extract, which is a heterologous plant.

The sea grape extract has an effect of inhibiting enzymes related to diabetes, an ability to protect pancreatic beta cells and insulin secretion, an effect to improve insulin sensitivity in fat cells, for example, glucose uptake ability, Is excellent in the effect of improving the blood glucose controlling ability, so that diabetes can be treated, improved or prevented.

In this respect, the sea grape extract may be an active ingredient of a composition for the treatment or prevention of diabetes, specifically, a pharmaceutical composition for the treatment or prevention of diabetes. The pharmaceutical composition refers to a medicament which is used for diagnosing, treating, alleviating, treating or preventing an illness in an animal including a human being.

The composition for treating or preventing diabetes comprising the sea grape extract as an active ingredient may contain 0.001 to 99.9% by weight or 0.1 to 99% by weight or 1 to 20% by weight of the sea grape extract, However, the present invention is not limited thereto, and the content of the sea grape extract may be suitably adjusted to a pharmaceutically effective amount or a desirable amount depending on the use of the composition and the method of use.

The composition for treating or preventing diabetes according to the present invention may contain the sea grape extract alone as an active ingredient and may further include a pharmaceutically acceptable carrier, excipient or diluent depending on the formulation, use method and purpose of use have.

The composition for treating or preventing diabetes may be applied directly to animals including humans. The animal is a group of organisms corresponding to plants. It mainly consumes organic matter as nutrients, and differentiates digestive organs, excretory or respiratory organs. Preferably, it is a mammal, more preferably a human.

The sea grape extract may be used alone as an active ingredient in a composition for treating or preventing diabetes, and may further include a pharmacologically acceptable or nutritionally acceptable carrier, excipient, diluent or sub ingredient.

In particular, when the sea grape extract is used as a medicine or used for medicinal or pharmaceutical purposes, the sea grape extract may be mixed with a pharmaceutically acceptable carrier or excipient according to a conventional method, or diluted with a diluent to be used .

More specifically, in addition to the above-mentioned active ingredients, there may be further added nutritional agents, vitamins, electrolytes, flavors, coloring agents, aggravating agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, Glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , Dextrin, calcium carbonate, dextrin, calcium carbonate, propylene glycol, liquid paraffin, and physiological saline. However, it is not limited thereto, and any conventional carrier, excipient or diluent may be used. The components may be added to the sea grape extract, which is the active ingredient, either independently or in combination.

The pharmacologically acceptable means physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorder, dizziness or the like when administered to an animal, preferably a human.

The composition for the treatment or prevention of diabetes may further comprise at least one selected from the group consisting of conventional fillers, extenders, binders, disintegrants, anticoagulants, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, Colloids, glycerin, fragrances, emulsifiers or preservatives, and the like. The components may be added to the composition for treating or preventing diabetes, either independently or in combination.

In addition, the composition of the present invention may further comprise a substance used for the treatment or prevention of diabetes, in addition to the active ingredient. Preferably, the added substance for treatment or prevention of diabetes is 0.1 to 20 parts by weight per 100 parts by weight of the composition for treating or preventing diabetes, or 100 parts by weight of the sea grape extract May be added in the range of 5 parts by weight to 200 parts by weight, respectively, but is not limited thereto.

In particular, when the sea grape extract is used as a medicine or used for medicinal or pharmaceutical purposes, the sea grape extract may be mixed with a pharmaceutically acceptable carrier or excipient according to a conventional method, or diluted with a diluent to be used .

More particularly, the pharmaceutical composition for treating or preventing diabetes comprising the sea grape extract as an active ingredient may further comprise at least one selected from the group consisting of a nutrient, a vitamin, an electrolyte, a flavoring agent, a colorant, a thickening agent, a pectic acid and a salt thereof, And a salt thereof, an organic acid, a protective colloid thickener, a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like.

The carrier, excipient or diluent may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acicia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, 1, selected from the group consisting of water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, physiological saline, But not limited thereto, and any conventional carrier, excipient or diluent can be used. The components may be added to the active ingredient, i.e., sea grape extract, either independently or in combination.

The pharmaceutical composition for treating or preventing diabetes comprising the sea grape extract as an active ingredient may contain a pharmaceutically effective amount of the active ingredient relative to the total weight of the composition, specifically 0.001% by weight to 99.9% by weight, 0.1% by weight to 99% by weight, and more preferably 1% by weight to 50% by weight. The pharmaceutically effective amount may be appropriately varied depending on the disease and its severity, age, body weight, health condition, sex, administration route and treatment period of the patient.

In addition, the content of the additional ingredient may be added in the range of 0.1% by weight to 20% by weight based on the total weight of the pharmaceutical composition for treating or preventing diabetes, but is not limited thereto.

When the pharmaceutical composition for treating or preventing diabetes mellitus containing the sea grape extract is used as an active ingredient, it may be formulated with conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, Preservatives, and the like, and they can be used either orally or parenterally.

When the composition for treating or preventing diabetes is used as a pharmaceutical composition, the composition can be administered either orally or parenterally. For example, the composition can be administered through various routes including oral, transdermal, subcutaneous, intravenous, or muscular.

In addition, the formulations of the compositions for the treatment or prevention of diabetes may be varied depending on the method of administration and may be formulated so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal, ≪ / RTI > Examples of the above-described formulations include granules, powders, syrups, liquids, suspensions, tablets, injections, injectable tablets, cataplasma, capsules, soft gelatine capsules or hard gelatine capsules.

The formulation may further contain an excipient such as an ordinary filler, an extender, a binder, a disintegrant, a surfactant, an anticoagulant, a lubricant, a wetting agent, a flavor, an emulsifier, an antiseptic, a sweetener, a fragrance or a preservative.

In general, solid dosage forms for oral administration include tablets, pills, soft or hard capsules, pills, POWDERS, and granules, The composition may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in the sea grape extract. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

In addition, liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water or liquid paraffin, which is a commonly used simple diluent, various excipients, for example, Wetting agents, sweetening agents, perfumes, preservatives, and the like.

In addition, the pharmaceutical composition for treating or preventing diabetes comprising the sea grape extract of the present invention as an active ingredient may be in a preferable form depending on the method of use. In particular, A method known in the art can be employed to formulate the drug. Examples of the specific formulations include granules, powders, syrups, liquids, suspensions, tablets, injections, main tablets, cataplasma, capsules, soft or hard gelatin capsules and the like.

Furthermore, the compositions of the present invention may be formulated using any suitable method known in the art or using methods disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA .

The dose of the composition can be suitably determined by those skilled in the art in consideration of the administration method, the age, sex and weight of the recipient, severity of the disease, the condition, the degree of absorption of the active ingredient in the body, the rate of inactivation and the drug to be used. For example, the sea grape extract may contain 0.0001 mg / kg body weight to 500 mg / kg body weight, 0.01 mg / kg body weight to 400 mg / kg body weight ) Or 1 mg / kg (body weight) to 300 mg / kg (body weight), and may be administered once a day or divided into several times. However, the dosage is not limited in any way to the scope of the present invention.

According to another embodiment of the present invention, there is provided a food composition for improving or preventing diabetes comprising the sea grape extract as an active ingredient. The food composition may be a functional food composition, specifically a health functional food for improving or preventing diabetes.

In the present specification, the term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, Food, beverage, food additive, and beverage additive.

The food may be, for example, various foods, beverages, gums, tea, a vitamin complex, a health functional food, and the like. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods (E.g., fruit and vegetable beverages, two oils, fermented beverages, etc.), natural seasoning (e.g., ramen soup, etc.).

The food, the health functional food, the beverage, the food additive and the beverage additive can be produced by a usual production method.

In the present invention, the health functional food refers to a food group imparted with added value to function and express the function of the food by physical, biochemical, biotechnological techniques, etc., or to control the bio-defense rhythm of the food composition, Means a food which is processed and designed so that the body control function related to restoration and the like is sufficiently expressed to the living body.

The health functional food may include a food-acceptable food supplementary additive, and may further include suitable carriers, excipients and diluents conventionally used in the production of a health functional food.

In the present invention, beverage means a general term for drinking or enjoying a taste, and is intended to include a health functional beverage.

The beverage is not particularly limited as long as it contains the sea grape extract as an active ingredient as an essential ingredient at the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages have.

Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and Xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably 5 to 12 g, per 100 ml of the food composition of the present invention. In addition, the composition of the present invention may be used for natural fruit juice, fruit juice drink, And may further contain pulp for the production of beverages.

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not so critical, but may be selected in the range of 0 to 20,000 parts by weight per 100 parts by weight of the sea grape extract of the present invention.

In the present invention, the health functional beverage includes a beverage group to which added value is added to function and express the function of the beverage by using a physical, biochemical, biotechnological technique or the like, a bio-defense rhythm control of the beverage composition, Means a beverage which is processed by being designed so that the body control function related to recovery and the like is sufficiently expressed to a living body.

The health functional beverage is not particularly limited to the ingredients other than the marine grape extract of the present invention as an essential ingredient in the indicated ratios and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages .

Examples of the natural carbohydrates include polysaccharides such as disaccharides such as monosaccharides such as glucose and fructose, maltose and sucrose, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol . The flavor may be a natural flavor such as tautatin or a stevia extract such as rebaudioside A or glycyrrhizin or a synthetic flavor such as saccharin, aspartame and the like. The amount of the natural carbohydrate to be added may generally be about 1 to 20 g, preferably 5 to 12 g per 100 ml of the food composition of the present invention.

In addition, the composition of the present invention may further contain flesh for the production of natural fruit juice, fruit juice drink, and vegetable drink.

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not critical, but may be selected in the range of 0 to 20 parts by weight per 100 parts by weight of the sea vine extract of the present invention.

In the present invention, the health functional beverage includes a beverage group to which added value is added to function and express the function of the beverage by using a physical, biochemical, biotechnological technique or the like, a bio-defense rhythm control of the beverage composition, Means a beverage which is processed by being designed so that the body control function related to recovery and the like is sufficiently expressed to a living body.

The health functional beverage is not particularly limited to the ingredients other than the marine grape extract of the present invention as an essential ingredient in the indicated ratios and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages .

Examples of the natural carbohydrates include polysaccharides such as disaccharides such as monosaccharides such as glucose and fructose, maltose and sucrose, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol . The flavor may be a natural flavor such as tautatin or a stevia extract such as rebaudioside A or glycyrrhizin or a synthetic flavor such as saccharin, aspartame and the like.

The amount of the natural carbohydrate to be added may generally be about 1 to 20 g, preferably 5 to 12 g per 100 ml of the food composition of the present invention. In addition, the composition of the present invention may further contain flesh for the production of natural fruit juice, fruit juice drink, and vegetable drink.

In addition, in the food composition for the purpose of preventing or improving diabetes, the amount of the sea grape extract may be 0.01% to 15% by weight of the total food, and the beverage composition may contain 0.02 g 5 g, preferably 0.3 g to 1 g.

The sea grape extract, which is an active ingredient of the composition of the present invention, was confirmed to have an excellent antidiabetic effect by in vitro test and cell test. As a result, it was found that beta-catenin of the pancreas The present inventors have confirmed that the present invention has a superior cell protection effect that significantly improves the survival ability of the present invention, improves the insulin secretion ability, improves the blood glucose control ability, and exhibits a remarkable antidiabetic effect as compared with other plant extracts.

In the food composition for preventing or improving diabetes comprising the sea grape extract as an active ingredient, the amount of the sea grape extract may be 0.01 wt% to 15 wt% of the total food weight. In addition, it may be contained at a ratio of 0.02 g to 5 g or 0.3 g to 1 g based on 100 ml of the sea grape extract beverage composition.

The sea grape extract as an active ingredient of the composition for the treatment or prevention of diabetes or the composition for the treatment or prevention of diabetes according to the present invention inhibits the death of the beta cells of the pancreas associated with the secretion of insulin and increases insulin secretion, Significantly improved anti-diabetic effect is recognized.

The sea grape extract of the present invention has an excellent effect of inhibiting the death of beta cells of the pancreas associated with the secretion of insulin and improving the glucose control ability in muscle cells and adipocytes and is therefore useful for the treatment, prevention or improvement of diabetes mellitus In addition, it can be used widely in the food industry including health functional foods related to diabetes treatment and prevention or prevention of diabetes, since safety is also recognized as a result of cytotoxicity test as well as no adverse effects unlike artificial compounds. .

FIG. 1 is a graph showing the cytotoxicity of extracts of sea grape (CLE) and octane (COE) according to an embodiment of the present invention. The vertical axis of each graph represents cell viability (cell viability , And the horizontal axis represents the control (Control) in which no sample is administered and the dose (μg / ml) of each extract sample. FIG. 1A shows RINm5F 1b shows the cell survival rate for RINm5F pancreatic? Cells according to the treatment amount of Oxea extract (COE), Fig. 1c shows the cell survival rate for pancreatic? Cells, and Fig. And the cell survival rate of L6 muscle cells.
FIG. 2 is a graph showing the results of confirming the antidiabetic effect of sea grape extract (CLE) and octane extract (COE) according to an embodiment of the present invention. The vertical axis of each graph represents the activity of each enzyme , And the horizontal axis represents the control (Control) in which no sample was administered and the dose (μg / ml) of each extract sample, and FIG. 2 (a) 2b shows inhibitory activity against DPP-IV activity according to the dose of each extract, and FIG. 2c shows inhibitory activity against PTTP1B activity according to the dose of each extract.
FIG. 3 is a graph showing the results of confirming the glucose absorption enhancing effect of the sea grape extract (CLE) and octane extract (COE) according to an embodiment of the present invention. (%)), And the abscissa indicates the control (Control) in which no sample is administered and the dose (μg / ml) of each extract sample. FIGS. 3a and 3c show the fat cells of sea grape extract (FIG. 3B) and muscle cells (FIG. 3D), and the results of confirming the glucose uptake ability in the muscle cells (FIG. 3C) .
FIG. 4 is a graph showing the results of confirming the protective effect of RINm5F cells of sea cucumber extract (CLE) and octane extract (COE) according to an embodiment of the present invention. And the abscissa represents the control (Control) in which no sample is administered and the dose (μg / ml) of each extract sample. FIG. 4a shows the result of administration of sea grape extract (CLE) FIG. 4B shows the result of administering COE extract.
FIGS. 5 to 8 are photographs and graphs showing the results of confirming the antidiabetic effect-related protein expression or phosphorylation-inducing ability of sea grape extract (CLE) and octene extract (COE) according to an embodiment of the present invention. (Adipocytes) and Fig. 7 (muscle cells) are photographs showing the result of immuno-staining to quantify the amount of each protein, and Fig. 6 (adipocytes) and Fig. 8 5 and 7 are control, the middle is the sea-vine extract administration group (250 μg / ml), the right is the insulin administration group (100 nM), and FIGS. 6 and 8 The vertical axis of each graph represents the degree of expression or phosphorylation of the back protein compared to the control group (fold of induction vs. control), while the horizontal axis represents the control (control) and sea grape extract (μg / ml ) And insulin (nM) ≪ / RTI >

Hereinafter, preferred embodiments of the present invention will be described. The following examples are intended to illustrate the present invention, but the scope of the present invention is not limited by the following examples.

Manufacturing example  1. Preparation of experimental material

1-1. Preparation of reagents

2-NBDG (2-N- (7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino) -2-deoxyglucose (Invitrogen, USA), Gly- , 4-nitrophenyl phosphate bis (cyclohexyl-ammonium) salt, bovine serum albumin (BSA), lipopolysaccaride (LPS), NaNO ₂ , thiazolyl blue tetrazolium bromide, Oil Red O, 2-mercaptoethanol, dexamethasone (DEX) and 3-isobutyl-1-methylxanthine (IBMX) were purchased from Sigma (St. Louis, USA).

Dulbecco's modified Eagle's medium (DMEM), newborn calf serum (FCS), fetal bovine serum (FBS), phosphate buffered saline (PBS), trypsin-EDTA and penicillin-streptomycin were purchased from Hyclone (Longan, USA).

The β-actin primary antibody and the anti-mouse secondary antibody were purchased from Sigma Aldrich (USA), and the β-actin primary antibody and the anti-mouse secondary antibody were purchased from Cell Signaling Technology (USA), pIRS, IRS, pAKT, AKT, pPI3K, PI3K, pGSK3β, GSK3β, Anti-rabbit secondary antibody was purchased from Jackson ImmunoResearch (USA). Nitrocellulose membrane was purchased from Bio-rad.

1-2. RINm5F  Cell culture

The rat pancreatic β cell line, RINm5F pancreatic β cells, was purchased from the American Type Culture Collection (ATCC, USA). RINm5F pancreatic β cells were cultured in RPMI 1640 medium containing 100 units / ml penicilin, 100 μg / ml streptomycin, 2.5 μg / ml amphotericin B and 10% FBS in a 5% CO ₂ and 37 ° C incubator.

1-3. L6 Muscle cell  culture

L6 muscle cells were purchased from the American Type Culture Collection (ATCC, USA). L6 muscle cells were cultured in DMEM medium containing 100 units / ml penicilin, 100 ㎍ / ml streptomycin and 10% FBS in a 5% CO ₂ and 37 ° C incubator. For the cultivation of the L6 muscle cells, the culture medium was subcultured while being supplemented with fresh medium every 2 days.

Differentiation of the cultured L6 muscle cells was performed as follows. First, after culturing in DMEM medium containing 10% FBS, it was changed to 80% to 90% confluent state, and the cells were changed to DMEM medium containing 2% FBS for 5 days. Lt; / RTI >

1-4. 3 T3 - L1  Cultivation of fat precursor cells

3T3-L1 adipose precursor cells were purchased from the American Type Culture Collection (ATCC, USA). 3T3-L1 adipose precursor cells were cultured in DMEM medium containing 100 units / ml penicilin, 100 ㎍ / ml streptomycin and 10% FCS at 37 ° C and 95% O2 and 5% Respectively. These 3T3-L1 adipose precursor cells were subcultured and supplemented with fresh medium every 2 days.

In order to differentiate the cultured 3T3-L1 adipose precursor cells into adipocytes, the cells were seeded at a density of 1 × 10 ⁵ cells / well in a 6-well plate. When the cells were 100% confluent, 5 ㎍ / ml insulin, 1 μM DEX , DMEM supplemented with 0.5 μM IBMX and 10% FBS was induced to induce differentiation (day 0). After 2 days from induction of differentiation, DMEM (10% FBS ) Medium. This was used for the experiment after 6 days (8 days) while replenishing with DMEM (10% FBS) medium every 2 days.

Manufacturing example  2. Sea grapes  Preparation of extract

Sea algae, ie sea grape samples, were purchased and used for commercialization of sea grapes collected in Okinawa, Japan. Also, the jade samples of the comparative group were prepared by natural drying for 3 days at 20 ° C to 30 ° C and ventilating conditions, and then pulverized with a pulverizer. The jade samples were directly sampled at Geumgwang Beach, Jeonnam Province.

The sea algae extract, ie, sea grape extract (CLE) or jade extract (COE), is prepared by putting 20 g of sea grape or jade crushed into an extraction container and immersing it in a water purifier at 15 ° C. for about 2 hours to remove the salt , And then removing the salt completely, adding 200 mL of 100% ethanol and extracting it for 24 hours.

Each of the extracts thus obtained was filtered under reduced pressure using a paper filter (Whatman, No. 2), concentrated by using a rotary vacuum evaporator after filtration, and then concentrated using a freeze dryer at -70 ° C To obtain an extract. Each of the final extracts was stored at -20 ° C and used for the experiment.

MTT assay was performed to evaluate cytotoxicity or apoptosis of seaweed extracts on RINm5F pancreatic? Or L6 muscle cells of the two seaweed extracts prepared above. RINm5F pancreatic β cells or L6 muscle cells were plated at 1 × 10 ³ cells / well in a 96-well plate and cultured for 24 hours. (MTT, 2 mg / dL) was added to each well. After 24 hours of incubation, each extract (CLE or COE) ml) was added to each well and reacted for 4 hours. The absorbance of the MTT solution was measured at 540 nm (Immuno Mini NJ-2300) after removing the MTT solution and adding DMSO (Sigma-Aldrich, USA) for 15 minutes. The measured absorbance was compared with a control (control) in which no sample was treated, and the relative values for the control were shown in Figs. 1A to 1C.

As shown in FIG. 1A and FIG. 1B, the cytotoxicity of Rinm5F pancreatic .beta. Cells of sea cucumber extract (CLE) and octane extract (COE) was examined in comparison with the control group to which no sample was added No cell death was observed up to the concentration of 250 μg / ml, and thus it was confirmed that no cytotoxicity was observed. In addition, as shown in Fig. 1C, the cytotoxicity of sea grape extract (CLE) to L6 muscle cells was found to be apoptotic up to a concentration of 250 μg / ml in all the extracts compared to the control without the sample , It was confirmed that the sea grape extract (CLE) was not cytotoxic to muscle cells.

Based on the results of the above experiment, it was determined that the concentration of 250 μg / ml was not cytotoxic and thus it was considered safe. To confirm the anti-diabetic effect of sea grape extract (CLE) Experiments were conducted.

Example  One. Sea grapes  Identification of diabetes prevention or improvement effects of extracts 1

In order to confirm the anti-diabetic effect of the two kinds of seaweed extracts prepared in the above-mentioned production examples, a cell-free assay, specifically inhibiting? -Glucosidase activity, DPP-IV activity inhibitory activity and PTTP1B Activity inhibition was confirmed.

Specifically, the inhibitory activity of? -Glucosidase activity was determined by adding 10 占 퐇 of the algae extract and 50 占 퐇 of enzyme (rat intestinal acetone powder) to each well of a 96-well plate for 5 minutes at room temperature. 50 μl of 5 mM substrate was added to the reacted wells and reacted at room temperature for 10 minutes. After 10 minutes, 50 μl of the termination reagent was added, and the absorbance was measured at 405 nm with a VICTOR ™ Multilabel Plate Reader (PerkinElmer, USA). The measurement results are shown in Fig. As shown in FIG. 2A, both the sea grape extract (CLE) and the oxe (COE) extract showed inhibitory effects on? -Glucosidase. However, when the same content was treated, the sea grape extract (CLE) . In particular, it was confirmed that the inhibitory effect was remarkably observed at a concentration of less than 250 μg / ml, which was confirmed as safe.

For inhibition of DPP-IV activity, 10 μl of mouse serum, 25 μl of the seaweed extract and 25 μl of 50 μM Gly-Pro-AMC were added to each well of a Black 96-well plate and reacted at 37 ° C. for 60 minutes After 60 minutes, 70 μl of 3 M acetic acid as a stopping reagent was added and the fluorescence was measured at an excitation wavelength of 355 nm and an emission wavelength of 460 nm of VICTOR ™ Multilabel Plate Reader (PerkinElmer, USA) Respectively. The results of the measurement are shown in FIG. 2B. As shown in FIG. 2B, both the sea grape extract (CLE) and the octave (COE) extract showed inhibitory effects on DPP-IV activity. However, when the same content was treated, sea grape extract (CLE) And showed high inhibition.

In addition, in the case of PTP1B activity inhibition ability, the degree of dephosphorylation was measured using p-NPP as a substrate. Specifically, a solution of 20 mM p-NPP (0.1 M NaCl, 1 mM EDTA, 50 mM citrate, pH 6.0, 1 mM DTT) and the sample (algae extract) were added to a solution of PTP1B (0.05 μg / The reaction was carried out at 37 ° C for 30 minutes, 10 μl of 10 N NaOH was added to terminate the reaction, and the absorbance at 410 nm was measured. The results of the measurement are shown in FIG. 2C. As shown in FIG. 2C, the extracts of sea grape (CLE) and octane (COE) showed inhibitory effects on PTP1B activity. However, the inhibitory effect of COE extract was very limited, while that of sea grape (CLE) In the case of the extract, the effect of improving the performance was confirmed as the content was increased.

Through the above experiments, it has been found that the sea cucumber extract (CLE) has remarkably excellent antidiabetic effect through the inferiority of the sugar-producing enzyme such as glucosidase or DPP-IV enzyme which lowers the insulin secretory hormone .

Example  2. Sea grapes  Identification of prevention or improvement of diabetes in extract 2

In order to confirm the antidiabetic effect of the two kinds of seaweed extracts prepared in the above Preparation Examples, experiments using the three kinds of cells cultured in the above Preparation Example were carried out.

2-1. Party Absorption capacity  Check the improvement

In order to confirm the effect of improving the insulin sensitivity of seaweed extracts, that is, the glucose uptake improvement effect of CLE extract, the effect of improving glucose uptake ability was examined in muscle cells and fat cells corresponding to tissue cells.

Specifically, in order to confirm the effect of CLE extract on pancreatic beta cells, beta cell protection effect of beta cells of rat pancreas was confirmed.

In the above preparation example, 3T3-L1 adipocytes differentiated from 3T3-L1 adipocytes and differentiated L6 muscle cells were added to the culture medium, cultured for 24 hours, and then the medium was removed 40 μM of 2- [N- (7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino] -2-deoxy glucose (2-NBDG, Invitrogen, Carlsbad, Calif. . After the addition of the 2-NBDG, the fluorescence was measured at 485 nm and 535 nm of excitation of a VICTOR3 ™ Multilabel Counter (PerkinElmer, USA) for 30 minutes, washed twice with PBS, and calculated as a ratio to the control value And is shown in Fig.

As shown in FIGS. 3A and 3B, the sea grape (CLE) extract (FIG. 3 a) in the 3T3-L1 adipocyte improves sugar uptake in a concentration-dependent manner while the COE extract (FIG. Was not found to improve sugar absorbability. Also, as shown in FIGS. 3c and 3d, the sea grape (CLE) extract (FIG. 3c) also improved the glucose uptake ability in a concentration-dependent manner in the L6 muscle cells, while the extract of COE (FIG. 3d) It has been confirmed that it does not improve sugar absorption ability

2-2. Confirmation of pancreatic beta cell protection effect

In order to confirm the anti - diabetic effect of seaweed extract on the insulin resistance of pancreatic beta cell protection, that is, the extract of sea grape (CLE), cell protection effect using pancreatic beta cells was confirmed.

More specifically, the rat pancreatic beta cell line RINm5F was cultured to a state close to confluence. The RINm5F pancreatic β cells were placed in a 96-well plate at 1 × 10 ⁵ cells / well, followed by overnight incubation. Subsequently, algae extract was added to each well and MTT (2 mg / ml) solution was added to each well for reaction for 1 hour. The MTT solution was removed and DMSO was added. After 15 minutes, absorbance was measured at 540 nm (Immuno Mini NJ-2300), and the result was shown in FIG. 4 as a ratio to the control value.

As shown in FIG. 4, in the pancreatic beta cell RINm5F, the cell extracts of sea cucumber (CLE) (FIG. 4A) were found to increase cell numbers of pancreatic beta cells in a concentration-dependent manner, 4b), although the treatment concentration was increased, the cell number of the pancreatic beta cells was not increased, and it was confirmed that there was almost no cell protection ability.

2-3. Anti-diabetic  Related protein On the ability to express  About Sea grapes  Check the effect of the extract

Western blotting was performed on each protein to confirm the expression ability or phosphorylation activity of p-IRS-1, p-Akt, p-PI3K, p-GSK3? And GLUT4, Respectively.

Specifically, the adipocytes derived from the lipid precursor cells and the cells induced by the muscle cells were dispensed at 1 × 10 ⁵ cells / well in a 96-well plate, and were then allowed to overnight. After the addition of sea cucumber extract (CLE) or insulin for 48 hours, the culture medium was removed, washed twice with PBS, and lysis buffer (PRO-PREP protein extraction solution, Intron Biotechnology) And dissolved at 4 占 폚 for 10 minutes. The dissolved cells were harvested and centrifuged at 13,000 rpm and 4 ° C for 20 minutes to recover the supernatant and then compared with the protein, specifically p-IRS-1, p-Akt, p-PI3K, p-GSK3β and GLUT4 Specifically, the amounts of IRS-1, Akt, PI3K, GSK3? And? -Actin were quantified.

To quantify the protein, 30 μg of the recovered supernatant was added to SDS loading buffer (60 mM Tris, 25% glycerol, 2% SDS, 0.5% 2-mercaptoethanol, 0.1% bromophenol blue) And electrophoretically transferred to a nitrocellulose membrane. Then, the membrane was added to a solution of TTBS (10 mM Tris, 100 mM NaCl, 0.1% tween 20) in which 5% unmodified milk powder was added, and the mixture was blocked at room temperature for 1 hour. (P-IRS1 1: 1000, IRS1 1: 1000, p-Akt 1: 1000, Akt 1: 1000, p-PI3K 1: 1000, PI3K 1: 1000, p -GSK3? 1: 1000, GSK3? 1: 1000, GLUT4 1: 1000 and? -Actin 1: 5000) for 2 hours and then washed three times with TTBS solution. Then, secondary antibody (goat anti mouse 1: 5000, goat anti rabbit 1: 5000) containing peroxidase was reacted for 1 hour. The detection of the antibody was visualized using a chemiluminescent substrate solution and analyzed with UVP (image acquisition and analysis software, Visionwork ^™ LS).

The results of the analysis are shown in FIGS. 5 to 8. FIG. P-Akt, p-PI3K and p-GSK3? In both of FIGS. 5 to 6 (muscle cells, L6) and FIGS. 7 to 8 (adipocytes, 3T3-L1) (CLE) extract was found to induce the phosphorylation of IRS-1, Akt, PI3K and GSK3?, And the expression of GLUT4 (Fig. 5E, Fig. 6E, Fig. 7E and Fig. 8E ).

According to the above results, the sea grape extract (CLE) activates the ability of insulin in the blood to bind to the insulin receptor in the blood, increases insulin receptor expression, increases insulin resistance or increases insulin sensitivity It was confirmed that the blood glucose control ability was improved through the blood. Specifically, the sea grape extract is activated by the activation of the insulin receptor substance (IRS) and the activation of the Akt-PI3K pathway, which is an insulin signal transduction system in the target cell, transport 4, GLUT4) and migration to the surface of the cell membrane, thereby activating the intracellular movement of glucose in the blood, thereby improving the blood glucose control ability of the blood.

Therefore, from the above results, the sea grape extract (CLE), unlike the extract of COE, which is a plant of the same genus, showed the inhibitory effect on diabetic induction enzymes, sugar absorption related to insulin sensitivity and pancreatic beta It has been found that the anti-diabetic effect is remarkably excellent by remarkably improving the cell protection ability and further enhancing the expression and phosphorylation of the transcription factor associated with the suppression of diabetes in the additional experiment. Thus, the sea grape (CLE) ≪ / RTI >

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (6)

Sea grapes ( Caulerpa lentillifera ) extract as an active ingredient for the treatment or prevention of diabetes. The method according to claim 1,
Wherein the sea grape extract is extracted with any one selected from the group consisting of alcohols having 1 to 5 carbon atoms, water, and mixtures thereof. 3. The method of claim 2,
Wherein the sea grape extract is extracted with ethanol or an aqueous solution of ethanol. Sea grapes ( Caulerpa lentillifera ) extract as an active ingredient. 5. The method of claim 4,
Wherein the sea grape extract is extracted with any one selected from the group consisting of alcohols having 1 to 5 carbon atoms, water, and mixtures thereof. 6. The method of claim 5,
Wherein said food composition is a health functional food.

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