KR101386691B1 - Composition for preventing or treating diabetes induced complication containing dicaffeoyl quinic acid derivatives - Google Patents

Abstract

Changiza extracts and / or dicafeoyl quinic acid derivatives related to the prevention and / or treatment and / or amelioration of diabetic complications, more specifically, Changiza extracts, and dicaffeoyl quinic acid derivatives of formula (1) Provided is a pharmaceutical composition or food composition comprising at least one selected from the group consisting of active ingredients. The composition is not only excellent in antioxidant activity, aldose reductase inhibitory activity and final glycation product inhibitory action, and because there is almost no side effect and safety problems because it is a component of natural extract, it is useful for the treatment, prevention or improvement of diabetic complications. Can be used.

Description

COMPOSITION FOR PREVENTING OR TREATING DIABETES INDUCED COMPLICATION CONTAINING DICAFFEOYL QUINIC ACID DERIVATIVES}

The present invention relates to the use of Changiza extract and / or dicafeoyl quinic acid derivatives for the prevention and / or treatment and / or amelioration of diabetic complications, more specifically, the Changiza extract, and the dicapoyl quinic acid of formula (dicaffeoyl quinic). acid) relates to a pharmaceutical composition or a food composition comprising at least one selected from the group consisting of derivatives as an active ingredient.

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, which is a path through which glucose is converted into fructose via sorbitol and is composed of two reaction systems, and two enzymes are involved.

More specifically, the polyol refers to an alcohol reduced by aldose reductase (AR) from aldose or ketose, and in general, when glucose enters the cell, hexokinase It is converted into 6-glucose phosphate and then broken down through the pathway.

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 the cell. 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 in 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 likely to lead to complications of tissues in which relatively high concentrations of sorbitol are accumulated, such as the lens and peripheral nerve.

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 the existing treatment for preventing or treating diabetic complications, which emphasized the glycemic control effect of simply lowering blood sugar, recently, the prevention or treatment of diabetic complications aimed at inhibiting the formation of aldose reductase and the final glycated product. Research is underway for development.

Aldose reductase inhibitors reported so far include 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.

The present inventors conducted a study in order to meet the above requirements, as a result, the Changjaza extract and the active ingredient compound having a structure of formula (I) separated from it inhibits the aldose reductase, the production of the final sugar oxide and sorbitol In addition to inhibiting the production, it is possible to control the oxidative stress caused by the antioxidant action, as well as confirmed that it is useful as a natural product-derived diabetic complication prevention and / or treatment that does not cause any problems, completed the present invention.

Accordingly, one example of the present invention is a pharmaceutical composition for preventing and / or treating diabetes complications comprising as an active ingredient at least one selected from the group consisting of Changiza extract and dicapoyl quinic acid derivative having the structure of Formula 1. To provide.

Another example provides a food composition for preventing and / or ameliorating diabetic complications comprising at least one selected from the group consisting of Changiza extract and a dicaffeoyl quinic acid derivative having the structure of Formula 1.

The inventors of the present invention provide ABSA (2,2'-Azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid)) free radical scavenging activity and DPPH ((1,1-Diphenyl-2-picrylhydrazyl)) scavenging activity. It was confirmed that the extract, and the derivative of 3,5-dicafeoyl quinic acid having the structure of Formula 1, each had an antioxidant effect, was excellent in inhibiting aldose reductase, inhibited the formation of final glycated product, and sorbitol production in red blood cells. It was confirmed that the inhibitory ability was remarkably excellent, and these extracts and / or compounds were judged to be excellent in the treatment, improvement or prevention effect of diabetic complications, and completed this invention.

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

An embodiment of the present invention contains as an active ingredient at least one selected from the group consisting of Changzai extract, a derivative of dicaffeoyl quinic acid having a structure of Formula 1, and a pharmaceutically acceptable salt of the derivative It relates to a pharmaceutical composition for preventing and / or treating diabetic complications:

[Chemical Formula 1]

Wherein,

R1 and R3 are hydrogen,

R2 and R4 is a cafe oil group,

R 5 is hydrogen or an alkyl group having 1 to 4 carbon atoms, preferably a methyl group.

The caffeoyl group has the structure of Formula 2 below:

(2)

In a preferred embodiment, the derivative of dicafeoyl quinic acid may be 3,5-dicafeoyl quinic acid (R 5 = hydrogen), or methyl-3,5-dicafeoyl quinic acid (R 5 = methyl group) More preferably methyl-3,5-dicafeoyl quinic acid (R 5 = methyl group).

Such pharmaceutically acceptable salts include both addition salts of acids or bases and stereochemically isomeric forms thereof. The salt includes any salt that maintains the activity of the parent compound in the subject to be administered and does not cause an undesirable effect, and is not particularly limited. Such salts include inorganic and organic salts, for example acetic acid, nitric acid, aspartic acid, sulfonic acid, sulfuric acid, maleic acid, glutamic acid, formic acid, succinic acid, phosphoric acid, phthalic acid, tannic acid, tartaric acid, hydrobromic Acids, propionic acid, benzenesulfonic acid, benzoic acid, stearic acid, ecylic acid, lactic acid, bicarboxylic acid, bisulfuric acid, bitartaric acid, oxalic acid, butyric acid, calcium edetic acid, campylic acid, carbonic acid, chlorobenzoic acid, citric acid, Edetic acid, toluenesulfonic acid, edicillic acid, ecylinic acid, fumaric acid, gluceptic acid, pamoic acid, gluconic acid, glycollylasanyl acid, methyl nitric acid, polygalatroxonic acid, hexylisorcinonic acid, mal Lonic acid, hydrabamic acid, hydrochloric acid, hydroyodoic acid, hydroxynaphthoic acid, isethionic acid, lactobionic acid, mandelic acid, estolinic acid, slime acid, napsilic acid, muconic acid, p-nitromethanesulfo Nic acid, hexamic acid, pantothenic acid, mono Hydrogen phosphoric acid, dihydrogen phosphoric acid, salicylic acid, sulfamic acid, sulfanilinic acid, methanesulfonic acid, theocyclic acid. Further, in the form of the basic salt, for example, alkali and alkaline earth metal salts such as ammonium salts, lithium, sodium, potassium, magnesium and calcium salts, for example benzine, N-methyl-D-glucamine, hydride Salts with an organic base, such as a labamine salt, and salts with amino acids, such as, for example, arginine, lysine. The salt form may also be converted to the free form by treatment with a suitable base or acid. “Additional salts” include compounds of formula I and solvates that salts thereof can form. Such solvent compounds may be, for example, hydrates, alcoholates and the like.

The changja is a fruit of the Asteraceae (Xanthium strumarium L.) or large tail, the oxtail is an annual herb widely grown in Korean fields and is distributed in Asia, Europe and North America such as Japan, Manchuria, China, Taiwan, and the Philippines. Chang-za has been reported to have analgesic effects on malignant tumors, and the concentrated liquid by boiling Chang-za is known to be effective for chronic rhinitis, arthritis and neuralgia. It is also reported that the water extract of Chang-za has an immune function activating effect.

Changchang extract used for the prevention and / or treatment of diabetic complications in the present invention may be prepared according to a conventional method for producing plant extracts, for example, extract obtained by extracting Changchang, Changchang dry matter, or Changchang extract as an extraction solvent. It may be water or a solvent fraction obtained by fractionation by adding a fractional solvent to the crude extract.

The extraction solvent may be at least one selected from the group consisting of water and an organic solvent. The organic solvent is a polar solvent containing a straight-chain or branched alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone and the like; Nonpolar solvents including ether, benzene, hexane, dichloromethane and the like; And it may be one or more selected from the group consisting of a mixed solvent of the polar solvent and non-polar solvent. The linear or branched 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 may be used at a concentration of 100% (v / v), or mixed with water, and used as an aqueous solution of alcohol at a concentration of 50 to 99.9% (v / v), for example, 75 to 99% (v / v). have.

In a preferred embodiment, the extractant may be at least one selected from the group consisting of water and a straight chain or branched alcohol having 1 to 5 carbon atoms, more specifically, water and straight chain or powder having 1 to 4 carbon atoms. It may be one or more selected from the group consisting of topographic alcohol, more specifically, may be water, methanol, or 75 to 99% (v / v) ethanol aqueous solution. The extraction may be performed at 4 ° C. to 120 ° C., specifically 50 ° C. to 90 ° C., more specifically 60 ° C. to 80 ° C., but is not limited thereto. The extraction time is not particularly limited, but may be 10 minutes to 12 hours.

In the present invention, the extraction method used for manufacturing Changja extract may be any conventional method of preparing plant extracts, and specifically, may be cold extraction, hot extraction, thermal extraction, ultrasonic extraction, etc., conventional extraction equipment, ultrasonic grinding extractor. Or a fractionator can be used.

In addition, the crude extract extracted with the extraction solvent is a group consisting of a straight chain or branched alcohol having 1 to 4 carbon atoms (eg, butanol, etc.), hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water, and the like. The fractionation process can be further roughened with one or more solvents. Two or more kinds of solvents may be used for the fractionation, and each solvent fraction may be used sequentially or in combination 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 crude extract prepared or the fraction obtained by performing the fractionation process may then be removed by performing a filtration, concentration, and / or drying process, it can be carried out both filtration, concentration and drying. Specifically, the filtration can be performed using a filter paper or a vacuum filter, and the concentration can be reduced by using a vacuum concentrator, for example, a rotary evaporator. The drying can be performed by, for example, freeze drying.

Changiza extract of the present invention, such as Changiza water extract and / or methanol extract and fractions thereof, and 3,5-dicapoyl quinic acid derivatives having a structure of formula (I), which are separated therefrom (and / or pharmaceuticals thereof) Acceptable salts) were confirmed through ABTS free radical scavenging activity and DPPH scavenging activity, and the antioxidant effect was recognized, which has the effect of treating, improving or preventing diabetic complications due to oxidative stress caused by increased autooxidation of glucose. As a result of measurement using animal lens, it has excellent aldose reductase, inhibits the progression of diabetic complications by polyol pathway and final glycation end products, and has excellent therapeutic effect of diabetic complications, and produces sorbitol in final glycation end products and erythrocytes. It was confirmed that it can inhibit the progression of diabetic complications.

Changjaza extract, which is an active ingredient of the composition for preventing or treating diabetic complications, and the 3,5-dicafeoyl quinic acid derivative having the structure of Formula 1 have excellent activity for preventing or treating diabetic complications, and can be used for food. Since it is manufactured using natural plants, side effects are not a problem, and safety is excellent.

The composition for preventing or treating diabetic complications of the present invention includes the Changja extract and / or a 3,5-dicafeoyl quinic acid derivative (and / or a pharmaceutically acceptable salt thereof) having the structure of Formula 1 as an active ingredient. In addition, it may further include a pharmaceutically acceptable carrier, excipient and / or diluent depending on the formulation, the method of use and the purpose of use.

In the present invention, the diabetic complication refers to the complication of diabetes mellitus, and the oxidative stress caused by an increase in the autooxidation of glucose and the final glycation product and glucose produced by the combination of high blood sugar and protein in the body due to abnormalities of the polyol pathway caused by hyperglycemia 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 examples 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 Can be.

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 preventing or treating diabetic complications may be directly applied 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.

Changjaza extract as an active ingredient in the pharmaceutical composition for preventing and / or treating diabetes complications of the present invention and / or 3,5-dicapoyl quinic acid derivative having a structure of Formula 1 (and / or a pharmaceutically acceptable salt thereof) The content of may be 0.001% to 99.9% by weight, specifically 0.1% to 99% by weight, and more specifically 1% to 20% by weight, but is not limited thereto. It can be used by appropriately adjusting according to the method.

As used herein, 'pharmaceutically acceptable' refers to a physiologically acceptable and, when administered to an animal, preferably a human, typically does not cause an allergic reaction such as gastrointestinal disorders, dizziness or similar reactions. It means not to. The pharmaceutically effective amount may be appropriately changed depending on the disease and its severity, the age, body weight, health condition, sex, administration route and treatment period of the patient.

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, propylene glycol, liquid paraffin, and physiological saline. However, the present invention is not limited thereto, and any conventional carrier, excipient or diluent may be used. In addition, the composition for preventing or treating 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 preventing or treating diabetic complications.

In addition, the composition of the present invention may further include a substance having a known preventive and / or therapeutic effect on diabetes complications in addition to the active ingredient.

Methods of administering the pharmaceutical composition for preventing and / or treating diabetic complications can be administered orally or parenterally, for example, oral, transdermal, subcutaneous, intravenous or intramuscular. The administration target of the pharmaceutical composition for preventing and / or treating diabetic complications may be a mammal including a human, and preferably, a human.

In addition, the formulation of the pharmaceutical composition for preventing and / or treating diabetic complications may vary depending on the method of use, and the present invention belongs to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be formulated using methods well known in the art. In general, solid dosage forms for oral administration include tablets, pills, soft or hard capsules, pills, powders and granules, which may contain one or more Excipients such as 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. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, Sweeteners, fragrances, preservatives, and the like.

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 any suitable method known in the art or using methods disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA .

The dosage of the composition can be determined in consideration of the administration method, the age, sex, the severity of the patient, the condition of the patient, the degree of absorption of the active ingredient in the body, the inactivation rate and the drug to be used together. 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, And may be administered once or several times in divided doses. The dose is not intended to limit the scope of the invention in any way.

In another embodiment, to prevent diabetic complications comprising at least one selected from the group consisting of Changiza extract, 3,5-dicafeoyl quinic acid derivatives having the structure of Formula 1, and pharmaceutically acceptable salts of the derivatives; Provides a food composition for improvement. The food composition for preventing and / or improving diabetic complications may further include appropriate 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.

As used herein, the term " food " means a natural product or a processed product containing one or more nutrients. Preferably, it means that the food can be directly eaten through a certain degree of processing. It is intended to include food, food additives, health functional foods and beverages.

Examples of foods that may include the Changja 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.

In the present invention, the functional food refers to a food group which is imparted with added value to function and express the function of the food by using physical, biochemical, biotechnological techniques and the like, the regulation of the biological defense rhythm of the food composition, Means a food which is processed and designed so that the body control function regarding the body is sufficiently expressed to the living body. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In the present invention, beverage is a generic term for drinking or enjoying a taste, and is intended to include functional beverages. The beverage is not particularly limited to other ingredients other than including the Changja extract as an active ingredient in the indicated ratio as an active ingredient, 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 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-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating 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 extract of the present invention, but is not limited thereto.

The functional beverage according to the present invention can be used to control the bio-defense rhythm of the beverage group or beverage composition to which the added value is imparted so that the function of the beverage acts on the specific purpose by physical, biochemical, biotechnological, Means a beverage which has been designed so that the body control function related to restoration and the like is sufficiently expressed in the living body.

The functional beverage is not particularly limited to other ingredients except for containing the Changja 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. 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 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 food composition for preventing and / or improving diabetic complications of the present invention, Changjaza extract as an active ingredient, the amount of the Changzai extract may be included as 0.00001% to 50% by weight of the total food weight, When the total volume of the food based on 100 ml may be included in a ratio of 0.001 g to 50 g, preferably 0.01 g to 10 g, but is not limited thereto.

Changza extract of the present invention, derivatives of 3,5-dicaffeoyl quinic acid having the structure of Formula 1, and / or pharmaceutically acceptable salts of the derivatives are ABTS free radical scavenging. As a result of the activity and DPPH scavenging activity, it was confirmed that the antioxidant effect was recognized, the aldose reductase inhibitory activity was excellent, and the final glycosylation inhibitory activity and the sorbitol and galactitol production inhibitory ability in erythrocytes were remarkably excellent. Not only has been found to be excellent in the treatment, improvement or prevention effect, but since it is derived from natural products and is unlikely to cause side effects, it can be used in various ways for the treatment, improvement and prevention of diabetic complications.

Figure 1 shows the results of HPLC chromatograms of the fractions and isolated compounds of the present invention, (1) ethyl acetate fraction of Changja metalol extract, (2) isolated 3,5-dicapoyl quinic acid, ( 3) shows the results for the isolated methyl-3,5-dicafeoyl quinic acid, respectively.

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 may 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 Changiza Extract and Separation of Compounds from It

1.1. Preparation of Changiza Extract

1.1.1. Changja Water Preparation of Extracts and Fractions

100 g of Changza ( Xanthium strumarium Linne ) was immersed in 1 L of water, and then extracted by hot needle extraction. More specifically, 1g of water was added to 100g of Changja and completely immersed, followed by extraction while refluxing at 100 ° C. to prepare Changja Water extract.

The obtained Changja water extract was sequentially fractionated into 3 L of methylene chloride, ethyl acetate (3 L) and butanol (3 L) in a separatory filter, and concentrated under reduced pressure in a water bath at 40 ° C., to give a methylene chloride fraction of the water extract, Ethyl acetate fractions, butanol fractions, and water fractions were prepared.

The obtained Changja water extract and fractions were concentrated under reduced pressure using a vacuum concentrator, and then freeze-dried at -20 ° C. The obtained extracts and fractions were stored frozen until used in the experiment.

1.1.2. Changja Methanol Extracts and Fractions

Methanol extract concentrated by depressurizing filtration of 4.5 kg of Changshaza ( Xanthium strumarium Linne ) in 5 L of 100% (v / v) methanol for 3 hours and filtration under reduced pressure. The filtrate was concentrated under reduced pressure in a 40 ° C water bath. The concentrate was suspended in 3 L of distilled water. The methanol extract suspended in distilled water was sequentially partitioned into 3 L of methylene chloride, ethyl acetate (3 L), butanol (3 L), and water (3 L) in an aliquot, and concentrated under reduced pressure in a 40 ° C. water bath. It was. The methylene chloride fraction, ethyl acetate fraction, butanol fraction, and water fraction of Changsha methanol extract were prepared.

1.2. Separation of compounds

After extracting 4.5 kg of Chang'anza ( Xanthium strumarium Linne ) with 5% of 100% (v / v) methanol for 3 hours (3 times), the resultant was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure in a water bath at 40 ° C. Was suspended in 3 L of distilled water. The methanol extract suspended in distilled water was sequentially partitioned into 3 L of methylene chloride, ethyl acetate (3 L), butanol (3 L), and water (3 L) in an aliquot, and concentrated under reduced pressure in a 40 ° C. water bath. It was. Among them, the ethyl acetate fraction was selected and suspended in 3.02 g with methanol solvent, coated with 7.5 g of Reverse Phase C ₁₈ , completely dried overnight, and then dissolved in 20% (v / v) methanol solvent. Separation was performed using 20% (v / v) to 50% (v / v) methanol solvent in RP C-18 column chromatography. Instrumental analysis As a result of Bruker Avance 600 NMR spectrometer (Karlsrube, Germany), 3,5-dicaffeoyl quinic acid and methyl-3,5-dicafeyl quinic acid (methyl-3, 5-dicaffeoyl quinic acid) was obtained (see FIG. 1; (1) HPLC chromatogram of ethyl acetate of Changza metalol extract, (2) HPLC chromatogram of isolated 3,5-dicafeoyl quinic acid, (3) HPLC chromatogram of isolated methyl-3,5-dicafeoyl quinic acid).

Methyl-3,5-dicafeoyl quinic acid: yellow powder form

Molecular Weight: 553 [M + Na] ⁺

¹ H-NMR: δ 2.19 (brd, J = 12.5 Hz), 1.97 (brd, J = 12.5 Hz), 5.37 (brdd, J = 9.5, 4.5 Hz), 3.95 (dd, J = 9.5, 3.0 Hz), 5.29 (dt, J = 7.0, 3.5 Hz), 2.19 (brd, J = 12.5 Hz), 1.97 (brd, J = 12.5 Hz), 6.24, 6.12 (d, J = 16.0 Hz), 7.48, 7.41 (d, J = 16.0 Hz), 7.04, 7.04 (d, J = 2.5 Hz), 6.77, 6.76 (d, J = 8.0 Hz), 6.99, 6.99 (d, J = 8.0 Hz)

¹³ C-NMR: δ 73.6 (C-1), 35.6 (C-2), 72.0 (C-3), 67.8 (C-4), 71.0 (C-5), 35.9 (C-6), 167.1 ( C-1 '), 166.4 (C-1 "), 115.9 (C-2'), 115.8 (C-2"), 146.8 (C-3 '), 146.7 (C-3 "), 126.7 (C- 4 '), 126.5 (C-4 "), 115.7 (C-5'), 114.8 (C-5"), 146.5 (C-6 '), 146.1 (C-6 "), 149.7 (C-7' ), 149.4 (C-7 "), 117.0 (C-8 '), 116.9 (C-8"), 122.5 (C-9'), 122.3 (C-9 "), 53.0 (OCH ₃ ), 174.8 ( CO).

3,5-Dicafeoyl quinic acid: white powder form

ESI-MS, m / z: 539.12 [M + Na] ⁺

¹ H-NMR: δ 2.21-2.34 (m, H-2), 5.47 (m, H-3), 4.01 (dd, J = 8.1, 3.1, H-4), 5.44 (m, H-5), 2.22-2.28 (m, H-6), 7.07 (d, J = 2.0, H-2 '), 6.79 (d, J = 8.1, H5'), 6.96 (d, J = 8.1, 2.0, H-6 '), 7.62 (d, J = 15.9, H-7'), 6.34 (d, J = 15.9 H-8 '), 7.07 (d, J = 2.0, H-2 "), 6.79 (d, J = 8.1, H-5 "), 6.94 (dd, J = 8.1, 2.0, H-6"), 7.60 (d, J = 15.9, H-7 "), 6.30 (d, J = 15.9, H-8" ).

¹³ C-NMR: 74.77 (C-1), 35.97 (C-2), 72.56 (C-3), 70.81 (C-4), 71.91 (C-5), 37.73 (C-6), 177.42 (C -7), 127.82 (C-1 '), 115.25 (C-2'), 146.51 (C-3 '), 149.34 (C-4'), 116.43 (C-5 '), 122.97 (C-6' ), 147.00 (C-7 '), 115.46 (C-8'), 168.87 (C-9 '), 127.70 (C-1 "), 115.14 (C-2"), 146.51 (C-3 "), 149.43 (C-4 "), 116.43 (C-5"), 123.04 (C-6 "), 147.19 (C-7"), 115.01 (C-8 "), 168.42 (C-9")

Example 2 Antioxidant Effect Measurement

In order to confirm the prevention or treatment of diabetic complications of the extracts, fractions and compounds prepared in Example 1, the antioxidant effects of the extracts, fractions and compounds were measured to determine the effect of inhibiting the development of diabetic complications due to stress. The antioxidant effect was confirmed by ABTS free radical scavenging activity. Dimethyl sulfoxide (DMSO) was added to the extract, each fraction or each compound obtained in Example 1 to prepare a solution at a concentration of 1 mg / ml, and used as a test solution in this antioxidant test.

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 test solution (extract, fraction or compound solution of 1 mg / ml concentration) and 290 μl of ABTS radical solution were added and reacted at room temperature for 10 minutes, and then absorbance was measured at 750 nm. In order to contrast with the ABTS radical inhibition rate of the Changza extract, a known antioxidant Trolox (sigma-aldrich) was used as a positive control. Radical scavenging activity of the extracts, fractions, and compounds of the present invention is shown in Tables 1 to 3 below in terms of IC ₅₀ values calculated by the concentration required to remove 50% of ABTS radicals.

Sample Name IC50 (uM) Trolox 4.67 3,5-di-O-caffeylquinic acid > 6.45 methyl-3,5-di-O-caffeylquinic acid 2.41

Sample Name IC50 (ug / ml) Trolox 3.89 Changiza Water Extract 22.96 Changja Water Extract-Ethyl Acetate Fraction 7.68 Changja Water Extract-Butanol Fraction 8.94

Sample Name IC50 (ug / ml) Trolox 4.67 Changiza Methanol Extract 127.89 Changiza Methanol Extract-Methylene Chloride Fraction 78.32 Changsha Methanol Extract-Ethyl Acetate Fraction 8.52 Changsha Methanol Extract-Butanol Fraction 9.34 Changja Methanol Extract-Water Fraction 84.47

As shown in Table 1 to Table 3, the fraction of Changzai water extract or methanol extract, and the compound 3,5-dicafeoyl quinic acid and methyl-3,5-dicapoyl quinic acid are equivalent to or more than the known antioxidant Trolox It was found to have antioxidant activity, and among these, methyl-3,5-dicafeoyl quinic acid was found to have very excellent antioxidant activity. From these results, it was confirmed that the Changza fraction and 3,5-dicafeoyl quinic acid and methyl-3,5-dicafeoyl quinic acid can control the induction of diabetic complications due to oxidative stress caused by autooxidation of glucose.

Example 3 Aldose Reductase Inhibitory Activity

In order to confirm the prophylactic or therapeutic effect of diabetic complications of the extracts, fractions and compounds prepared in Example 1, the inhibitory effect of aldose reductase inhibition of each compound was confirmed to inhibit the onset of diabetic complications by aldose reductase inhibition. . In order to measure the inhibitory capacity of the aldose reductase, the lens of the rat was extracted and used. DMSO was added to the extract, each fraction or each compound obtained in Example 1 to prepare a solution at a concentration of 1 mg / ml, and used as a test solution in the present aldose reductase inhibitory effect test.

Experimental animals were used to receive five rats 10 weeks old male WKY (Wistar Kyoto) from the central laboratory animals (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% (v / v) with ammonium sulfate (ammonium sulfate), the centrifuged supernatant was again taken, and again 70% (v / v) Ammonium sulfate was added to the mixture to stir for 1 hour, and the pellets obtained by centrifugation were suspended in a minimum amount of buffer, followed by dialysis for 1 day, and then used as an enzyme source.

100 μl of the enzyme source, 700 μl of phosphate buffer (pH 7.0) as a reaction buffer, 100 μl of DL-glyceraldehyde (0.1 M) as a substrate, and 100 μl of NADPH (1.6 mM) as a coenzyme. After reacting by adding DMSO (dimethyl sulfoxide) to the extract obtained in Example 1, each fraction or each compound to the reaction solution as a solution of 1 mg / ml concentration, the rate of decrease of NADPH absorbance at 340 nm was measured by spectrophotometer. It was. In order to check the aldose reductase inhibitory ability of each of the extracts, fractions and compounds, a known aldose reductase inhibitor Quercetin was used as a positive control. The aldose reductase inhibitory ability of each extract, fraction and compound of the present invention is shown in Tables 4 to 6 by converting the concentration at the time when aldose reductase activity is inhibited by 50% to IC ₅₀ value.

IC ₅₀ (uM) 3,5-di-O-caffeylquinic acid 0.19 methyl-3,5-di-O-caffeylquinic acid 0.30 Quercetin (positive control) 7.42

IC ₅₀ (ug / ml) Changiza Water Extract 3.51 Changja Water Extract-Ethyl Acetate Fraction 0.82 Changja Water Extract-Butanol Fraction 1.95 Quercetin 7.42

Sample Name IC50 (ug / ml) Quercetin 1.65 Changiza Methanol Extract 2.32 Changiza Methanol Extract-Methylene Chloride Fraction 2.64 Changsha Methanol Extract-Ethyl Acetate Fraction 0.37 Changsha Methanol Extract-Butanol Fraction 0.59 Changja Methanol Extract-Water Fraction 1.1

In addition, the same method as the experiment using the lens of the rat using human aldose reductase cDNA clone obtained by recombination of human aldose reductase cDNA clone from 21C Frontier Human Gene Bank in Daejeon, KIST, Gangneung The results obtained by performing are shown in Tables 7 and 8 below.

IC ₅₀ (uM) 3,5-di-O-caffeylquinic acid 1.34 methyl-3,5-di-O-caffeylquinic acid 0.67 Quercetin (positive control) 10.99

Sample Name IC50 (ug / ml) Quercetin 3.32 Changiza Methanol Extract 4.81 Changiza Methanol Extract-Methylene Chloride Fraction  - Changsha Methanol Extract-Ethyl Acetate Fraction 1.63 Changsha Methanol Extract-Butanol Fraction 2.34 Changja Methanol Extract-Water Fraction -

As shown in Table 4 to Table 8, Changja water extract, methanol extract and fractions thereof, and compound 3,5-dicafeoyl quinic acid and methyl-3,5-dicapoyl quinic acid are all known aldose reductions. It was found to have higher aldose reductase inhibitory activity compared to Quercetin, an enzyme inhibitor. As a result, the extracts, fractions and compounds of the present invention were found to have an excellent acid effect in the treatment or improvement of diabetic complications caused by abnormal polyol pathway.

Example 4 Inhibition Effect of Final Glycation Product Production

In order to confirm the prevention or treatment of diabetic complications of the extracts, fractions and compounds prepared in Example 1, diabetic complications by inhibiting the final glycation product production by measuring the inhibitory effect of the final glycation product production of the extracts, fractions and compounds Onset inhibition effect was confirmed. DMSO was added to the extract, each fraction or each compound obtained in Example 1 to prepare an extract at a concentration of 10 mg / ml, and used as a test solution of the final glycation product production inhibition test.

To 50 ml of 0.1 M sodium phosphate buffer (pH 7.4), add 2.5 g of bovine serum albumin (BSA) as a protein source and 38.46 ul of 5 mM methyl glyosal as a sugar source, and 0.02 Antibacterial sodium azide was added at a concentration of% (w / v) to prevent bacterial formation during the reaction. After 30 μl of the test solution was added to 1.5 ml of the mixed reaction solution, the reaction solution was reacted at 37 ° C. for 5 days, followed by fluorescence spectrophotometer (Excitation (Ex): 350 nm, Emission (EM): 450 nm). ) And the results are shown in Table 9 and Table 10 below. In order to check the inhibitory ability of the final glycation product production of the test solution, Aminoguanidine, a known final glycation product production inhibitor, was used as a positive control.

Sample Name IC50 (uM) Aminoguanidine (positive control) 1410 3,5-di-O-caffeylquinic acid > 193.80 methyl-3,5-di-O-caffeylquinic acid 112.69

Sample Name IC50 (ug / ml) Aminoguanidine (positive control) 69.75 Changja Water Extract-Ethyl Acetate Fraction 58.16 Changja Water Extract-Butanol Fraction 140.62

As shown in Table 9 and Table 10, Changzai water extract and fractions thereof, and compounds 3,5-dicafeoyl quinic acid and methyl-3,5-dicafeoyl quinic acid are known amino acid production inhibitors Aminoguanidine It was confirmed to have a high final glycation product production inhibitory ability in comparison with As a result, the extracts, fractions, and compounds of the present invention were found to have an excellent effect on the treatment or improvement of diabetic complications caused by the production of the final glycation oxide.

Example 5 Galactitol Formation Inhibitory Activity

In order to confirm the prophylactic or therapeutic effect of each extract, fraction, and compound of the diabetic complications prepared in Example 1, to measure the inhibitory ability of galactitol production of each extract, fraction, and compound to develop diabetes complications by the type galactitol production inhibitory ability Inhibitory effect was confirmed. DMSO was added to the extract, each fraction or each compound obtained in Example 1 to prepare a solution at a concentration of 1 mg / ml, and used as a test solution in this galactitol production inhibitory effect test.

The galactitol production inhibitory ability was used to receive five 10-week-old male WKY (Wistar Kyoto) from a central laboratory animal (Korea). The experimental animals were anesthetized with ether, and the fertilizer was extracted. Then 15 ml of 15% (v / v) FBS was added to 35 ml of TC-199 medium (Sigma-aldrich). Thereafter, 100 units / ml penicillin, 0.1 mg / ml streptomycin, and 30 mM galactose were added to prepare a media mixture. 1 ml of the media compound and 50 μl of the prepared test solution were added to the extracted lens. The media was changed every 24 hours and incubated at 37 ° C. under 5% CO ₂ for 6 days. After 6 days, 150ul of phosphate buffer was added to the lens and ground. Thereafter, centrifugation was performed at 11,000 g for 10 minutes. The supernatant was then filtered through a filter.

70 μl of the filtered supernatant was used for benzoylation. Specifically, 20 μl of 1M KH ₂ PO ₄ , 3 μl of benzoyl chloride and 6 μl of 8M NaOH were added to 70 μl of the neutralized sample (supernatant), followed by vortexing and mixing, followed by 1.4 MH _3. 10 μl of PO ₄ was added for neutralization, 100 μl of ethyl acetate was added thereto, and vortexed and mixed, followed by centrifugation using a centrifuge to completely divide up and down. After removing the supernatant with nitrogen, 25 μl of acetonitrile and a mixture of water (70:30 (v / v)) was dissolved and absorbance was measured at 288 nm.

In order to contrast the galactitol production inhibitory ability of each compound, Quercetin, a known galactitol production inhibitor, was used as a positive control, and was used without adding a test solution or Quercetin as a control. The galactitol production inhibitory ability of each compound of the present invention was measured using the measured absorbance, and is shown in Table 11 below.

Galactitol content (ug / lens wet weight) Control 982 Quercetin 188 3,5-di-O-caffeylquinic acid 153 methyl-3,5-di-O-caffeylquinic acid 117

As shown in Table 11, Quercetin, a known inhibitor of galactitol production, showed 80.86% galactitol production inhibitory ability, whereas 3,5-dicafeoyl quinic acid was 84.42% and methyl-3,5-dicafeoyl quinic. Acid was found to exhibit 88.09% galactitol production inhibitory activity. From the results of the galactitol production inhibitory test results, each compound of the present invention exhibits the inhibitory ability of galatitol production beyond the known inhibitors, thereby providing an excellent treatment or improvement effect of diabetic complications caused by abnormal polyol pathway and final glycoxide production. It was confirmed to have.

Claims (4)

A derivative of dicaffeoyl quinic acid having a structure of Formula 1, and a pharmaceutically acceptable salt of the derivative, containing at least one selected from the group consisting of active ingredients,
Selected from the group consisting of diabetic retinopathy, diabetic cataract, diabetic keratosis, diabetic nephropathy, diabetic neuropathy, diabetic peripheral neuropathy, atherosclerosis and diabetic angiopathy,
Pharmaceutical compositions for preventing or treating diabetic complications:
[Chemical Formula 1]

Wherein,
R1 and R3 are hydrogen,
R2 and R4 is a cafe oil group,
R 5 is an alkyl group having 1 to 4 carbon atoms.
The method of claim 1,
The dicafeoyl quinic acid derivative is methyl-3,5-dicafeoyl quinic acid (methyl-3,5-dicaffeoyl quinic acid),
Pharmaceutical composition for preventing or treating diabetes complications. delete delete

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