KR20110087420A - Composition for preventing or improving diabetes induced complication containing anthemis tinctoria extract - Google Patents

Abstract

PURPOSE: A composition containing Anthemis tinctoria extract for preventing or treating diabetes complication is provided to suppress glycation end product with less side effects and safety. CONSTITUTION: A composition for preventing or treating diabetes complication contains Anthemis tinctoria extract as an active ingredient. The Anthemis tinctoria extract is prepared by isolating Anthemis tinctoria flower using water, alcohol of 1-5 carbon atoms, or mixture thereof. The diabetes complication is diabetic retinopathy, cataract, diabetic cornea, diabetic nephropathy, diabetic neuropathy, diabetic peripheral nerve failure, or atherosis. A food composition for preventing or treating diabetes complication contains the Anthemis tinctoria flower extract as an active ingredient. The Anthemis tinctoria extract is contained in 0.001-99.9 weight%. The composition for preventing or treating diabetes complication is taken by oral or parenteral administration.

Description

COMPOSITION FOR PREVENTING OR IMPROVING DIABETES INDUCED COMPLICATION CONTAINING ANTHEMIS TINCTORIA EXTRACT}

The present invention relates to a pharmaceutical composition comprising as an active ingredient a plant extract having an effect of treating or preventing diabetes complications.

Diabetes is a chronic disease in which glucose in the blood, ie, blood sugar, is higher than normal and releases glucose into the urine, resulting from abnormal insulin metabolism or abnormal physiological activity in β-cells of the Langerhans island in the pancreas. It is one of the representative adult diseases. In advanced countries, the number of diabetics is increasing every year, and in Korea, the number of diabetics is gradually increasing due to the improvement of living standards and westernized lifestyle. Recently, about 5% of the total population or at least 2.5 million people are estimated to be diabetics, and diabetes is the fourth leading cause of death among Koreans.

In the long-term hyperglycemic environment caused by diabetes, high osmotic stress caused by hyperglycemia or glycated products invade the kidneys, nerves, retina and large and small blood vessels of the whole body can cause a variety of complications. Because of the greater risk of complications caused by diabetes, more efforts are being made today to suppress, prevent or improve the incidence or progression of diabetes complications (diabetes complications) than the treatment of diabetes itself. Diabetes complications are long-standing diabetes mellitus, usually due to chronic complications after 10-15 years after the onset of diabetes. Types of chronic complications are diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy. Various diseases have been reported, including.

Diabetes complications are those diseases in which glycated products invade the kidneys, nerves, retinas, and large and small blood vessels of the whole body due to a prolonged period of high blood sugar levels caused by diabetes. For example, diabetic retina, which is the leading cause of blindness in adults, Disease and the like. Mechanisms leading to diabetic complications include osmotic stress caused by changes in the mechanism of non-enzymatic glycation of protein and polyol pathway, and oxidative stress caused by free radicals. And the like have been reported.

The polyol pathway is one of the glucose metabolic pathways, in which glucose is converted into fructose via sorbitol and consists of two reaction systems, and two enzymes are involved. More specifically, a polyol is an aldose or kerosene. It means alcohol reduced from ketose by aldose reductase (AR). In general, when glucose enters the cell, it is converted into 6-phosphate glucose by hexokinase and then degraded through the corresponding pathway. do.

When the concentration of glucose increases, hexokinase that binds to it is saturated, and excess glucose binds to aldose reductase, which is less affinity than hexokinase, and synthesizes sorbitol, thereby accumulating sorbitol in cells. Sorbitol is highly polarized and difficult to diffuse out of the cell membrane.

Two enzymes, aldose reductase and sorbitol dehydrogenase, are involved in the sorbitol synthesis process. These enzymes are present in the tissues of the body, including peripheral nerves, retina, cornea, iris, lens, kidney, and red blood cells. Especially, aldose reductase is used for retina, sclera, lens, kidney, red blood cells, brain, muscle, liver and peripheral nerves. It exists on the back.

The aldose reductase is a reaction limiting enzyme that converts glucose into sorbitol using NADPH as a coenzyme as an enzyme that causes the first reaction of the sorbitol synthesis process. Once sorbitol is produced, sorbitol is converted to fructose by sorbitol dehydrogenase.

Under physiological conditions, the flow of synthesis by these two enzymes is in equilibrium with most tissues. Therefore, the accumulation of sorbitol is almost negligible in most tissues.Also, aldose reductase affinity for glucose is lower than hexokinase and most intracellular glucose is phosphorylated, resulting in low sorbitol production rate. do. However, in the case of diabetic patients, if the concentration of glucose in the blood increases further, aldose reductase is activated to excessively reduce glucose to sorbitol, and sorbitol excessively produced by the aldose reductase is sorbitol dehydrogenase Is converted to fructose, and since fructose is about 10 times faster than non-enzymatic glycosylation of protein, high concentration of fructose is combined with protein, resulting in advanced glycosylation end-product, AGE) to accelerate the formation.

In particular, the production of such end glycosylated products may be more problematic in peripheral nerves such that glucose permeation is not affected by insulin. Cellular permeation of glucose in tissues that do not depend on insulin is dependent on the glucose concentration present in the cells in tissues that do not depend on insulin because the intracellular glucose uptake, including the peripheral nerves, is associated with peripheral nerves, lens, retina, cornea, and blood vessels. When intracellular vineyard concentration of insulin-independent tissues such as renal glomeruli or erythrocytes increases, glucose is promoted by aldose reductase, an essential enzyme of the polyol pathway, which is present in large amounts in the cells of these tissues, resulting in excessive accumulation of sorbitol. This can lead to increased osmotic pressure, resulting in edema, necrosis, and collapse of myelin sheath, and increased osmotic pressure can disrupt the osmotic control system, leading to the introduction of water, leading to diabetic complications. The diabetic complications include diabetic cataracts, diabetic retinopathy, diabetic keratosis, diabetic neurosis, and diabetic nephropathy, depending on the site where sorbitol accumulates.

In addition, activation of the aldose reductase may exacerbate oxidative stress and result in a deficiency of intracellular miyoinositol and taurine. This results in intracellular deficiency of miyoinositol by oxidative stress either directly down-regulating the Na ⁺ -myoinositol cotransporter and taurien transporter, or indirectly through interfering with sodium-slope by sodium-potassium adenosine triphosphatase (ATPase). This is a promising mechanism for complications in tissues in which relatively high concentrations of sorbitol, such as the lens and peripheral nerves, accumulate.

Despite the high concentration of sorbitol in these tissues, aldose reductase is continuously expressed to further accumulate sorbitol. Therefore, various metabolic processes, electrophysiological changes, reduction of energy consumption in neural tissues, changes in neural tissues, etc. occur in neural tissues. In addition, the increase in osmotic pressure caused by the accumulation of sorbitol and fructose by aldose reductase and sorbitol dehydrogenase promotes water inflow into lens fibers, resulting in cell swelling.

This whereas increasing the permeability of the material that is being held properly in the lens, K ⁺ in the crystalline lens, an amino acid (amino acid), glutathione (glutathione), Inositol (inositol) and ATP concentration decreases of, Na ^+, Cl ^- The concentration of ions gradually increases. As this process proceeds, the increased two ions cause electrolyte change, causing secondary osmotic change, which in turn causes lens swelling. Due to this osmotic change, the cell membrane of the lens increases permeability to almost all substances except large proteins of a certain size or more, resulting in dense nuclear cataract.

As described above, the lower K ⁺ / Na ⁺ ratio in the lens than normal can reduce protein synthesis, so the K ⁺ / Na ⁺ ratio can be an indicator of diabetic complications. In addition, the concentration of glucose-related metabolites (sorbitol) and mediator enzymes in red blood cells may be an indicator of progression of indirect diabetic complications. According to clinical studies, concentration ratios of sorbitol content in erythrocytes, sorbitol content in red blood cells, and glucose content in plasma were significantly higher in diabetic patients compared to normal patients, and these indicators could be used as a diagnostic method for diabetes. It became.

The diabetic retinopathy or nuclear hardening cataract, which is one of diabetic complications, is known to be caused mainly by the intracellular accumulation of sorbitol. In particular, in patients with insulin-independent diabetes, the progression of diabetic retinopathy has a greater effect on the duration of the abnormal sorbitol concentration than the sorbitol content in the red blood cells, so that the sorbitol content of the diabetic patients is higher than normal. Clinical studies have reported that early use of aldose reductase inhibitors may be effective in the prevention and treatment of diabetic retinopathy.

In addition, the final glycation end products have been reported to activate aldose reductase, the main enzyme of the polyol pathway in human microvascular endothelial cells. Generally, aldose reductase has a very low affinity for glucose at steady state, but at high concentrations of glucose, aldose reductase is activated to excessively reduce glucose to sorbitol, and the sorbitol is converted to fructose by sorbitol dehydrogenase. Is switched. As described above, since fructose is about 10 times faster than glucose in glucose, high concentrations of fructose bind to protein and eventually form final glycation products. Accelerate

The non-enzymatic glycosylation of the protein refers to a condensation reaction, that is, a miliad reaction, between an amino acid group such as a lysine residue of the protein and a reducing sugar without enzymatic action, resulting in the final glycation product.

The non-enzymatic glycosylation of the protein comprises: 1) an amino acid group, such as a lysine residue of the protein, and an aldehyde or ketone of the reducing sugar undergo a nucleophilic addition reaction without enzymatic action to form a Schiff base, which is a product of an early stage, Schiffbase and adjacent ketoamine adducts condensate with each other to produce a reversible Amadori type of early glycosylated product, and 2) high blood sugar condition persists, so that reversible Amadori type of early glycated product is not degraded and rearranged. (rearrangement), it can be divided into the step of producing an irreversible final glycosylated product by cross-linking with the protein.

Unlike the Amadori-type early glycation products, which are the products of the reversible reaction, the final glycation products are irreversible reaction products. Once produced, blood sugar is not decomposed even when blood sugar returns to normal, but accumulates in the tissues during protein survival. Change abnormally. When glycosylation occurs in proteins such as the basement membrane, plasma albumin, lens protein, fibrin, and collagen by non-enzymatic protein glycosylation, the resulting glycosylated product abnormally changes the structure and function of the tissue, thereby diabetic retinopathy. ) Can cause chronic diabetes complications such as cataract, diabetic nephropathy, diabetic neuropathy or atherosclerosis.

In addition, the increase in oxidative damage or oxidative stress caused by hyperglycemia occurs when hyperglycemia increases the production of free oxygen radicals or decreases the antioxidant mechanisms, which vary widely and autooxidation of glucose. It is believed that the increase of free radicals due to the increase of), the increase of non-enzymatic glycosylation and self-oxidation thereof, and the change of redox state due to activation of the polyol pathway.

The self-oxidation of glucose means that the glucose itself is enolized to reduce oxygen molecules and make oxides. With regard to the production of free radicals by increasing the self-oxidation of glucose, the following problems are expected. When the self-oxidation of the glucose proceeds due to hyperglycemia, superoxide ions (O ² −), hydroxy radicals (OH), hydrogen peroxide (H ₂ 0 ₂ ), and the like are formed, which are crosslinked, segmented, and the like. It has been reported to cause lipid peroxidation or damage protein. It is also reported that free radicals promote the formation of end glycosylated products, which in turn can promote autooxidation.

As such, the polyol pathway, non-enzymatic glycosylation and oxidative stress mechanisms are linked to each other to cause diabetic complications. Therefore, in order to delay, prevent or treat the development of diabetic complications, simply controlling blood glucose is insufficient, and it is very important to inhibit the formation of aldose reductase and final glycation product. Therefore, in the study of existing diabetic complications for treating or preventing diabetes, which emphasizes the glycemic control effect of simply lowering blood sugar, recently, the treatment or prevention of diabetic complications aimed at suppressing the formation of aldose reductase and the final glycated product. Research is underway for development.

The aldose reductase inhibitors reported so far are sorbinil, alrestatin, epalrestat, ponalestat, tolrestat, and aminoguanidine-HCl (aminoguanidine-HCl) as a protein glycosylation inhibitor. Since the aldose reductase inhibitor binds to aldose reductase, it inhibits the conversion of glucose to sorbitol, thereby preventing the reduction of miyoinositol and the damage of sodium-potassium ATPase, and at a different position from the location that interacts with glucose. Reacts with dose reductase to produce noncompetitive inhibition of aldose reductase. In addition, the aminoguanidine is a nucleophilic hydrazine (bin) with the amadori product to prevent cross-linking with the protein to inhibit the production of the final glycated product can be delayed or prevented the development of complications.

Although many kinds of inhibitors including the aldose reductase inhibitors and protein glycosylation inhibitors have been developed and synthesized, there is a problem that toxicity is induced upon long-term administration, and thus, the search and development of new safer inhibitors are desired.

In order to meet the above requirements, the present invention inhibits aldose reductase, inhibits the production of the final sugar oxide, and can control the oxidative stress caused by antioxidant action, as well as from the natural products that do not cause side effects An object of the present invention is to provide a composition for treating or preventing diabetes complications.

In order to achieve the above object, the present invention provides a composition for treating or preventing diabetic complications comprising a natural product, that is, a plant-derived active ingredient. More specifically, the present invention provides a composition for treating or preventing diabetic complications comprising Dias chamomile extract as an active ingredient.

The inventors of the present invention, the Dias chamomile bark extract is confirmed by the ABTS free radical scavenging activity and DPPH scavenging activity is excellent in the antioxidant effect, aldose reductase inhibitory activity and final glycation product production inhibitory ability, sorbitol production in red blood cells It was confirmed that the inhibitory ability was remarkably excellent, not only was it proved to be excellent in the treatment, improvement or prevention effect of diabetic complications, but also derived from natural products and confirmed that it is unlikely to cause side effects, thus completing the present invention. .

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

The present invention relates to a composition for treating or preventing diabetic complications comprising Dias chamomile extract as an active ingredient.

Dyer's Chamomile, Anthemis tinctoria ) is a perennial plant of the Asteraceae family, also called yellow chamomil, and is native to Europe. Yellow flowers bloom at the end of the stem, mainly used as a raw material for dyeing. In the West, it is called Dyer's Chamomile. Dias chamomile is a sunshine-like, cold-resistant, cold-growing plant that grows to about 50 cm in height, and the flower is yellow and blooms from June to July. A well drained and sunny place grows well regardless of soil, strong against pests but weak against aphids. The flower of Dias chamomile is used as a natural mordant and is mainly cultivated to obtain a raw material of the dye, and also used as an ornamental or a fragrance.

The Dias chamomile extract may be prepared according to a conventional method for producing a plant extract, for example, the flowers, leaves, stems, roots or seeds of Dias chamomile or extracts thereof, preferably the flowers of Dias chamomile It may be prepared by adding a solvent or fractionation by adding a fractional solvent to the crude extract prepared by extraction with an extraction solvent.

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 alcohol having 1 to 5 carbon atoms, alcohol dilution water, ethyl acetate or acetone and a non-polar solvent of ether, chloroform, benzene, hexane or dichloromethane, or a mixed solvent of the polar solvent and non-polar solvent. . The alcohol having 1 to 5 carbon atoms may be methanol, ethanol, propanol, butanol, isopropanol, and the like, but is not limited thereto. In addition, the alcohol dilution water may be one obtained by diluting the alcohol in water at 50 to 99.9% (v / v).

Extraction solvent of the Dias chamomile extract of the present invention is preferably at least one selected from the group consisting of water, alcohol having 1 to 5 carbon atoms, alcohol dilution water and mixtures thereof, more preferably water, 1 to It may be any one selected from the group consisting of alcohols and mixtures thereof, and even more preferably 75 to 99% aqueous ethanol solution. For example, the extraction process may be performed at 4 ° C. to 120 ° C., or 50 ° C. to 90 ° C., or 60 ° C. to 80 ° C., but is not limited thereto. In addition, the extraction time is not particularly limited, but may be 10 minutes to 12 hours.

Dias chamomile extract of the present invention may be prepared according to a conventional method for producing a plant extract, specifically may be cold extraction, hot extraction, heat extraction, ultrasonic extraction, etc., conventional extraction equipment, ultrasonic grinding extractor or A fractionator can be used.

In addition, the extract extracted with the solvent may be further subjected to the fractionation process with any one solvent selected from the group consisting of hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water and mixtures thereof. Two or more solvents may be used in the fractionation, and each solvent extract may be prepared using sequentially or mixed according to the polarity of the solvent. In addition, the fractionation process may be performed at 4 ° C. to 120 ° C., or 50 ° C. to 90 ° C., or 60 ° C. to 80 ° C., but is not limited thereto.

The prepared extract or the fraction obtained by performing the fractionation process can be filtered or concentrated or dried to remove the solvent, it can be carried out both filtration, concentration and drying. Specifically, the filtration may be performed using a filter paper or a reduced pressure filter, the concentration may be concentrated under reduced pressure using a reduced pressure concentrator, for example, a rotary evaporator, and the drying may be performed by, for example, a lyophilization method.

Dias chamomile flower ethanol extract of the present invention is confirmed through the ABTS free radical scavenging activity and DPPH scavenging activity, the antioxidant effect is recognized, the treatment, improvement or prevention effect of diabetic complications due to the oxidative stress caused by the increase of autooxidation of glucose The result of the measurement using the lens of the experimental animal is excellent in aldose reductase, inhibits the progression of diabetic complications by the polyol pathway and the final glycation end products, has an excellent therapeutic effect of diabetic complications, It has been confirmed that the production of sorbitol in erythrocytes can be inhibited to block the progression of diabetic complications.

Diaus chamomile extract is an active ingredient of the composition for the treatment or prevention of diabetic complications not only has excellent activity for treating or preventing diabetic complications, but is prepared using a natural plant that can be used for food, so side effects are not a problem. Excellent safety

The composition for treating or preventing diabetic complications of the present invention may include the Dias chamomile extract alone as an active ingredient, and further includes a pharmaceutically acceptable carrier, excipient or diluent according to the formulation, method of use and purpose of use. can do.

In the present invention, diabetic complication refers to the complications of diabetes mellitus, oxidative stress due to an increase in the autooxidation of glucose and the final glycation product and glucose produced in the body due to abnormalities of the polyol pathway caused by hyperglycemia, and the combination of high blood sugar and protein in the body Means a disease caused by. The diabetic complications include diabetic vascular disorders, neuropathy and infectious diseases.

The diabetic vascular disorders cause arteriosclerosis of relatively thick blood vessels such as aorta, coronary arteries, and lower extremity, resulting in diseases such as heart attack, angina pectoris, and stroke, and small blood vessels such as retina and kidney (glomeruli) and capillaries. It damages micro-vessels such as blood vessels or changes blood circulation, causing glomerulosclerosis, retinopathy, renal failure, vision disorders, lower extremity gangrene. The diabetic neuropathy is a neurological disorder caused by the above causes. Representative peripheral nerve disorders due to abnormalities of carbohydrates and lipid metabolism are typical, peripheral nerve disorders, loss of keyboard, paralysis of motor nerves, autonomic nerve disorders, etc. Infectious diseases include pyelonephritis, pyoderma, pericardial folliculitis, eczema and candidiasis.

Representative diseases of the diabetic complications are any one selected from the group consisting of diabetic retinopathy, diabetic cataract, diabetic keratosis, diabetic nephropathy, diabetic neuropathy, diabetic peripheral neuropathy, atherosclerosis and diabetic vascular complications It can be one.

Diabetic retinopathy is one of the microvascular complications and is a serious complication that causes blindness in diabetic patients. The diabetic nephropathy is one of microvascular complications and is a disease caused by sclerotic lesions of renal glomerular capillaries.

The composition for treating or preventing diabetic complications may be directly applied to animals including humans. The animal is a biological group corresponding to a plant, which mainly consumes organic matter as nutrients, and means that the digestive organ, excretory canal, and respiratory tract are differentiated, preferably a mammal, more preferably a human.

The Dias chamomile extract may be used alone as an active ingredient in a composition for treating or preventing diabetic complications, and may further include other pharmacologically acceptable carriers, excipients, diluents or accessory ingredients. More specifically, when the Dias chamomile extract is used as a medicament, or for medicinal or pharmaceutical use, the Dias chamomile extract is mixed with a pharmaceutically acceptable carrier or excipient according to a conventional method or diluted with a diluent. It can be used diluted.

In this case, the content of the Dias chamomile extract in the composition may be 0.001% to 99.9% by weight, 0.1% to 99% by weight or 1% to 20% by weight, but is not limited thereto. And according to the method of use, the content of the Dias chamomile extract may be used by appropriately adjusting the pharmaceutically effective amount or the preferred amount.

The term 'pharmaceutically acceptable' means that when administered to a physiologically acceptable animal, preferably a human, usually does not cause an allergic reaction, such as gastrointestinal disorders, dizziness or similar. The pharmaceutically effective amount may be appropriately changed depending on the disease and its severity, the patient's age, weight, health condition, sex, route of administration and duration of treatment.

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, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , Dextrin, calcium carbonate, propylene glycol, liquid paraffin, and one or more selected from the group consisting of physiological saline, but is not limited thereto, and all common carriers, excipients or diluents can be used. In addition, the composition for treating or preventing diabetic complications is conventional fillers, extenders, binders, disintegrants, anti-coagulants, lubricants, wetting agents, pH regulators, nutrients, vitamins, electrolytes, alginic acid and salts thereof, pectic acid and salts thereof, protection Sex colloids, glycerin, fragrances, emulsifiers or preservatives; The components may be added independently or in combination to the composition for treating or preventing diabetic complications.

In addition, the composition of the present invention may further include a substance used for the treatment or prevention of known diabetes complications in addition to the active ingredient.

When the composition for treating or preventing diabetic complications is used as a medicament, the administration method may be oral or parenteral, and for example, may be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

In addition, the formulation of the composition for treating or preventing diabetic complications may vary depending on the method of use, and is well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be formulated using the method. Generally, solid preparations for oral administration include tablets, pills, soft or hard capsules (CAPSULES), pills (PILLS), powders (POWDERS) and granules (GRANULES), and the like. Excipients, for example, may be prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Oral liquid preparations include suspending agents (SUSTESIONS), liquid solutions, emulsions (EMULSIONS) and syrups (SYRUPS) .Such as simple diluents such as water and liquid paraffin, various excipients such as wetting agents, Sweeteners, fragrances, preservatives and the like can be included.

Forms for parenteral administration are CREAM, LOTIONS, ONITMENTS, PLASTERS, LIQUIDS AND SOULTIONS, AEROSOLS, FRUIDEXTRACTS, Elixir (ELIXIR), INFUSIONS, SACHET, PATCH or INJECTIONS.

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

The dosage of the composition may be determined in consideration of the method of administration, age, sex, severity, condition of the patient, absorption of the active ingredient in the body, inactivation rate and the drug used in combination, for example, based on the daily effective ingredient 0.1 mg / kg (body weight) to 500 mg / kg (body weight), 0.1 mg / kg (body weight) to 400 mg / kg body weight or 1 mg / kg body weight to 300 mg / kg body weight It can be administered, and can be administered once or divided into several times. The dosage does not limit the scope of the invention in any aspect.

In addition, the present invention provides a food composition for improving or preventing diabetic complications comprising the Dias chamomile extract as an active ingredient. Food composition for improving or preventing diabetic complications comprising the Dias chamomile extract as an active ingredient may further comprise suitable carriers, excipients and diluents commonly used in the preparation thereof.

Examples of the food composition of the present invention include food, food additives, beverages or beverage additives.

In the present specification, the term "food" means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It is intended to include all foods, food additives, health functional foods and beverages.

Examples of foods that may contain the Dias chamomile extract include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food in the present invention includes special nutritional products (e.g., crude oil, infant food, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g., ramen, noodles, etc.), health supplements, seasoned foods (E.g., soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages (Eg, fruits, vegetable drinks, soy milk, fermented beverages, ice creams, etc.), natural seasonings, vitamin complexes, alcoholic beverages, alcoholic beverages and other health supplements. The food, beverage or food additive may be prepared by a conventional production method.

Functional food in the present invention is the control of biological defense rhythm, disease prevention and recovery of food groups or food compositions that have added value to the food by using physical, biochemical, biotechnological techniques, etc. It means a food processed and designed to fully express the gymnastics function related to the living body. The functional food may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.

In the present invention, the drink is a generic term for drinking to quench thirst or to enjoy the taste and is intended to include a functional drink. The beverage contains the Dias chamomile extract as an essential ingredient in the indicated ratios as an active ingredient, and there are no special limitations to the other ingredients, and may include various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. have. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and Sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate may generally be about 1 to 20 g, preferably 5 to 12 g per 100 ml of the food composition of the present invention. It may further contain pulp for preparation.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. These components can be used independently or in combination. The additive may be 0 to 200 parts by weight, preferably 0.00001 to 150 parts by weight, per 100 parts by weight of the Dias chamomile extract of the present invention, but is not limited thereto.

Functional beverage in the present invention is a biological defense rhythm control, disease prevention and the like having a beverage group or a beverage composition that has added value to the beverage by using physical, biochemical, biotechnological techniques, etc. to function and express the function of the beverage to a specific purpose Means a beverage that is designed and processed to fully express the gymnastics function related to recovery.

The functional beverage is not particularly limited to other ingredients except for containing the Dias chamomile extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and xylitol Sugar alcohols such as sorbitol and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The natural carbohydrate may be included in an amount of 0 to 20 parts by weight, preferably 1 to 18 parts by weight, more preferably 5 to 12 parts by weight, per 100 parts by weight of the composition of the present invention.

In addition, in the diabetic complication improvement or prevention food composition comprising the Dias chamomile extract of the present invention as an active ingredient, the amount of the Dias chamomile extract may be included as 0.00001% to 50% by weight of the total food weight, drink The composition may be included in a ratio of 0.001 g to 50 g, preferably 0.01 g to 10 g, based on 100 ml of the total food volume, but is not limited thereto.

As a result of confirming through ABTS free radical scavenging activity and DPPH scavenging activity, Dias chamomile extract of the present invention is recognized as an antioxidant effect, aldose reductase inhibitory ability, the final glycosylation inhibitory activity and sorbitol generation inhibition in red blood cells is remarkable In addition, it has been found to be excellent in the treatment, improvement or prevention of diabetic complications, and since it is derived from natural products, it is less likely to cause side effects. Can be used.

Hereinafter, the present invention will be described in detail with reference to Examples. However, the following examples are merely illustrative of the present invention and can be modified in many different forms, which are intended to illustrate the present invention, but the scope of the present invention is not limited by the following examples.

Example 1 Preparation of Dias Chamomile Extract

Dias chamomile 100 g of tinctoria ) flowers were immersed in 10 L of ethanol, and extracted with a warm needle extraction method.

More specifically, 10 L of 90% ethanol aqueous solution was added to 100 g of the Dias chamomile flower, and the Dias chamomile flower was completely immersed in the extraction solvent, and then extracted while refluxing at 70 ° C. Dias chamomile extract prepared by the extraction was concentrated under reduced pressure using a vacuum concentrator, respectively, and then lyophilized at -20 ℃. The obtained extract was stored frozen until used in the experiment.

Example 2 Determination of Treatment or Prevention Effect of Diabetic Complications 1-Measurement of Antioxidant Effect

In relation to the diabetic complication treatment or prevention effect of the Dias chamomile extract prepared in Example 1, in order to measure the inhibitory effect of the development of diabetic complications due to oxidative stress, the antioxidant effect of the Dias chamomile extract was measured. The antioxidant effect was confirmed by ABTS free radical scavenging activity and DPPH scavenging activity. In the experiment for confirming the following antioxidant effect, the Dias chamomile extract of the present invention was prepared by using methanol to the extract obtained in 1 mg / ml concentration for use in the experiment.

The ABTS free radical scavenging activity was measured by mixing 2 mM ABTS and 3.5 mM potassium persulfate, dissolving in 10 ml of distilled water, diluting 30 times with 290 ml of tertiary distilled water, and overnight. ) To induce ABTS radical production. 10 μl of the Dias chamomile extract prepared in Example 1 and 290 μl of the ABTS radical solution were added and reacted at room temperature for 10 minutes, and then the absorbance was measured at 750 nm. In order to check the ABTS radical inhibition rate of the Dias chamomile extract, a known antioxidant Trolox was used as a control. Radical scavenging activity of the Dias chamomile extract of the present invention is calculated by the concentration necessary to 50% scavenging ABTS radicals in terms of IC ₅₀ value, it is shown in Table 1 below.

In addition, the DPPH scavenging activity was measured by mixing 180 μl of DPPH dissolved in HPLC grade methanol and 30 μl of the extract at a concentration of 0.32 mM and reacting at room temperature for 20 minutes, and then absorbance was measured at 570 nm. In order to check the DPPH radical inhibition rate of the Dias chamomile extract, a known antioxidant Ascorbaic acid was used as a control. Radical scavenging activity of the Dias chamomile extract of the present invention is calculated by the concentration necessary to 50% scavenging DPPH radicals, and converted into IC ₅₀ values, and is shown in Table 2 below.

Concentration (ug / ml) ABTS radical inhibition rate (%) IC ₅₀ (ug / ml) Dias Chamomile Extract 33.33 58.99 28.10 16.67 30.29 3.33 16.30 Trolox 3.33 63.49 2.62 1.67 31.09 0.33 12.03

Concentration (ug / ml) DPPH radical inhibition rate (%) IC ₅₀ (ug / ml) Dias Chamomile Extract 142.86 52.17 135.85 71.43 27.14 14.29 4.18 Ascorbic acid 14.29 77.53 9.45 7.15 31.69 1.43 13.04

As shown in Table 1 and Table 2, Dias chamomile extract was confirmed to have antioxidant activity. From the above results, the Dias chamomile extract of the present invention was recognized that the antioxidant effect, it was confirmed that it can control the induction of diabetic complications due to oxidative stress caused by autooxidation of glucose.

< Example  3> Diabetes complications treatment or prevention effect Measurement 2 - Aldos  Reductase inhibitory activity

In connection with the diabetic complication treatment or prophylactic effect of the Dias chamomile extract prepared in Example 1, in order to measure the inhibitory effect of the onset of diabetic complications by the inhibition of aldose reductase, Measured. Inhibition of the aldose reductase was used to extract the lens of the rat. In the experiment for confirming the following aldose reductase inhibitory effect, the Dias chamomile extract of the present invention was prepared for the use in the experiment, the extract was prepared by adding DMSO to the extract obtained at a concentration of 1 mg / ml.

Experimental animals were used to receive five 10-week-old male WKY (Wistar Kyoto) from the central laboratory animals (South Korea). After the animal was anesthetized using ether, the lens was extracted from the experiment animal, and a certain amount of sodium phosphate buffer (pH 6.2) was added and ground to hybridize (homogenization) according to the wet weight of the extracted lens. After centrifuging the hybridized lens at 4 ℃, the supernatant was taken up to 40% saturated with ammonium sulfate, the centrifuged supernatant was again taken, and ammonium sulfate was added to 70% again for about 1 hour. After stirring, the pellets obtained by centrifugation were suspended in a minimum amount of buffer, followed by dialysis for 1 day, which was then used as an enzyme source.

100 μl of the enzyme source, 700 μl of phosphate buffer solution (pH 7.0) and 100 μl of DL-glyceraldehyde (0.1 M) as a substrate and 100 μl of coenzyme NADPH (1.6 mM) are prepared. After adding the enzyme solution to one reaction solution and reacting, the rate of decrease of NADPH absorbance at 340 nm was measured with a spectrophotometer. In order to check the aldose reductase inhibitory ability of the Dias chamomile extract, Quercetin, a known aldose reductase inhibitor, was used as a control. The aldose reductase inhibitory ability of the Dias chamomile extract of the present invention is shown in Table 3 below by converting the concentration at the time when the aldose reductase activity is inhibited by 50% to an IC ₅₀ value.

Concentration (ug / ml) Aldose reductase inhibitory ability (%) IC ₅₀ (ug / ml) Dias Chamomile Extract 10 96.58 4.88 5 53.11 One 12.95 Quercetin 5 77.01 2.40 One 45.08 0.5 20.01

As shown in Table 3, Dias chamomile extract was confirmed to have a similar aldose reductase inhibitory ability compared to Quercetin, a known aldose reductase inhibitor. From the result that the aldose reductase inhibitory ability is excellent, it was confirmed that the Dias chamomile extract of the present invention will be excellent in the treatment or improvement of diabetic complications caused by abnormal polyol pathways and the production of the final glycosylation.

< Example  4> Treatment or prevention effect of diabetic complications Measurement 3 - Final Glycation Product  Formation inhibitory effect

In connection with the diabetic complication treatment or prophylactic effect of the Dias chamomile extract prepared in Example 1, in order to measure the inhibitory effect of the development of diabetic complications by the inhibition of the final glycation product production, the final glycation product inhibitory effect Was measured. In the experiment to confirm the inhibitory effect of the final glycation product production, the Dias chamomile extract of the present invention is prepared for use in the experiment, the extract obtained by the addition of DMSO to the extract obtained in Example 1 to prepare an extract at a concentration of 1 mg / ml It was.

After dissolving a solution of 50 mg / ml protein source bovine serum albumin (BSA) and sugar source methyl glyosal 5 mM in 0.1 M sodium phosphate buffer (pH 7.4), Sodium azide, an antimicrobial agent, was added at a concentration of 0.02% to prevent bacterial formation during the reaction. After adding Dias chamomile extract to the reaction solution for 5 days at 37 ℃, using a fluorescence spectophotometer (fluorescence spectrophotometer) (Excitation (Ex): 350nm, Emission (EM): 450nm) by measuring The results are shown in Table 4 below. In order to contrast the final glycation end product production inhibitory ability of the Dias chamomile extract, Aminoguanidine, a known end glycation end product production inhibitory substance was used as a control.

Concentration (ug / ml) Final Glucose Formation Inhibition (%) IC ₅₀ (ug / ml) Dias Chamomile Extract 196.09 12.16 - Aminoguanidine 196.08 82.45 110.66 98.04 46.71 19.61 13.91

As shown in Table 4, Dias chamomile extract was confirmed to have the ability to inhibit the end glycation product production. It is expected from the result that the final glycosylated product inhibiting ability of the Dias chamomile extract of the present invention will be excellent in the treatment or improvement effect of diabetic complications caused by the production of the final glycoxide.

< Example  5> Treatment or prevention effect of diabetic complications Measurement 4 -Sorbitol production inhibitory activity

In connection with the diabetic complication treatment or prevention effect of the Dias chamomile extract prepared in Example 1, in order to measure the inhibitory effect of the development of diabetic complications by the sorbitol production inhibitory ability, the sorbitol production inhibitory ability of the Dias chamomile extract was measured. In the experiment for confirming the effect of inhibiting the sorbitol production, the Dias chamomile extract of the present invention was prepared in 1 mg / ml concentration by adding DMSO to the extract obtained in Example 1 for use in the experiment.

The sorbitol production inhibitory ability was used to receive five 10-week-old male WKY (Wistar Kyoto) from a central laboratory animal (Korea). The animals were anesthetized with ether, and then the abdomen was cut to collect blood from the heart. 1 mL of 2.2% sodium citrate was mixed with 9 mL of blood and slowly mixed up and down, and then centrifuged at 4 ° C. and 2,000 × g for 5 minutes. After centrifugation, the upper layer was discarded and washed three times with brine.

The red blood cells (RBC) obtained by the above method were treated with Kerbs-Ginger bicarbonate buffer and Dias chamomile extract, and reacted in a 5% CO ₂ incubater at 37 ° C. for 1 hour. After the reaction was completed, erythrocytes were washed three times with cold saline, added with cold 6% perchloride acid and centrifuged for 10 minutes at 4 ℃ and 2,000 xg conditions. The supernatant obtained after the centrifugation was neutralized by adding cold 2.5MK ₂ CO ₃ .

70 μl of the neutralized supernatant was used for benzoylation. Specifically, 20 μl of 1M KH ₂ PO ₄ , 3 μl benzoyl chloride and 8M NaOH were added to 70 μl of the neutralized sample, mixed by vortexing, and then 10 μl of 1.4 MH ₃ PO ₄ was added thereto. After neutralization, 100 μl ethyl acetate was added, vortexed and mixed, followed by centrifugation to completely divide the upper and lower parts. After removing the supernatant with nitrogen, it was dissolved in acetonitrile and a mixture of water (70:30) and absorbance was measured at 288 nm.

In order to check the sorbitol production inhibitory ability of the Dias chamomile extract, Quercetin, a known sorbitol production inhibitor, was used as a control. The sorbitol production inhibitory ability of the Dias chamomile extract of the present invention is shown in Table 5 below using the measured absorbance.

Concentration (ug / ml) Inhibition of intracellular sorbitol production (%) Dias Chamomile Extract 10 62.88 Quercetin 10 25.86

As shown in Table 5, Dias chamomile extract was confirmed to have a superior sorbitol production inhibitory ability, specifically about 250% inhibition when confirmed based on the same content compared to known sorbitol production inhibitor Quercetin. . From the results that the sorbitol production inhibitory ability is excellent, it was confirmed that the Dias chamomile extract of the present invention is excellent in the treatment or improvement of diabetic complications caused by abnormal polyol pathways and the production of the final glycosylation.

Claims (4)

A composition for treating or preventing diabetic complications comprising Dias chamomile extract as an active ingredient. The method of claim 1,
The Dias chamomile extract is a composition for treating or preventing diabetic complications that extracts one or more selected from the group consisting of water, alcohol having 1 to 5 carbon atoms, and mixtures thereof. The method of claim 1,
The diabetic complication is any one selected from the group consisting of diabetic retinopathy, diabetic cataract, diabetic keratosis, diabetic nephropathy, diabetic neuropathy, diabetic peripheral neuropathy, atherosclerosis and diabetic vascular complications Therapeutic or prophylactic composition. A food composition for improving or preventing diabetic complications, comprising, as an active ingredient, a dias chamomile flower extract obtained by extracting at least one selected from the group consisting of water, diol chamomile flowers, alcohols having 1 to 5 carbon atoms, and mixtures thereof as an extraction solvent.