KR20050003668A - Composition comprising an extract of Cuscuta species for preventing and treating diabetes - Google Patents

Abstract

PURPOSE: Provided is a composition comprising the extract of Cuscuta species which has excellent reducing effect on blood glucose level. Therefore, the composition has preventing and treating effects on diabetes and its complications. CONSTITUTION: The pharmaceutical composition for treating diabetes and its complications is characterized by containing the extract of Cuscuta species as an active ingredient, pharmaceutically acceptable carrier, diluent, or excipient. The extract is obtained by using water, C1-4 lower alcohol or their mixture as an extracting solvent.

Description

Composition comprising an extract of Cuscuta species for preventing and treating diabetes}

The present invention relates to a composition for preventing and treating diabetes, including extract of Tosa.

Diabetes is a metabolic disorder of multiple etiologies that is characterized by chronic hyperglycemia due to insulin secretion, insulin action, or a defect in both. When blood glucose levels are abnormally high for a long time, various complications occur due to chronic metabolic disorders and chronic blood vessel damage. Diabetes complications can be divided into acute and chronic. Acute complications include hypoglycemia, ketoacidosis, hyperosmotic non-ketone hyperglycemia, hyperglycemic coma, diabetic acidemia, hypoglycemia, and chronic complications include diabetic retinopathy and diabetic nephropathy. , Diabetic neuropathy, cardiovascular complications, and viral infections.

There are two types of diabetes, type I diabetes, insulin dependent diabetes (IDDM) and type II diabetes, non-insulin dependent diabetes (NIDDM). IDDM is caused by a deficiency of insulin, a glucose-regulating hormone in the blood, and most commonly occurs in infants and adolescents, called juvenile diabetes, and is considered an autoimmune disease. IDDM is generally treated with insulin, and other methods for treating IDDM disclose sulfated glycolipids in WO 92/19633, glycolipid complexes in WO 97/42974, and galactosyl in WO 99/33475. Ceramides, glucosylceramides and lactosylceramides are disclosed. Recently, transplantation of pancreatic or insulin-producing pancreatic cells has been used to address the underlying insulin deficiency.

NIDDM is the most common form of diabetes, mainly after the age of 40 and called adult diabetes. It accounts for about 90% of all diabetics and is a metabolic disorder caused by not producing enough insulin or using it properly. The cause of NIDDM is not clear, but both genetic and environmental factors, such as obesity and lack of exercise, are thought to work. NIDDM causes both insulin secretion in pancreatic cells and insulin action defects (insulin resistance) in target cells, but it is not yet clear which changes are of primary importance.

In diabetes, severe hyperglycemia occurs due to increased activity of glucose-6-phosphatase enzymes in the liver and reduced glucokinase activity. Even in this process, the liver cannot store them in the form of intrahepatic glycosides, but continuously produces glucose and releases it into the blood. In addition, diabetes also promotes gluconeogenesis, resulting in poor regulation of glucose metabolism resulting in various liver tissue changes. In NIDDM, large droplet fatty liver is common, which increases the migration of free fatty acids from the adipose tissue to the liver due to insulin deficiency and excess of glucagon and is neutral. This is due to the increase in triglycerides. In addition, while the use of glucose is reduced, glycogen breakdown and glucose neosynthesis are increased to promote fatty liver production.

NIDDM is usually first treated with diet and exercise therapy, and if the therapeutic effect by this method is insufficient, generally several types of drugs are used. Drugs that promote the release of insulin from the pancreas, or drugs that increase insulin sensitivity to make insulin work efficiently, or directly increase blood glucose levels by increasing glucose production in the liver or by increasing glucose uptake in muscles Drugs that affect and drugs that inhibit the absorption of sugar in the intestine. In many cases insulin is also used.

Insulin is essential for all IDDM patients, and NIDDM is also required for patients whose blood sugar is not controlled by diet and oral hypoglycemic agents. However, since insulin is a protein, it is hydrolyzed and inactivated in the digestive tract, so that it cannot be consumed orally and must be injected intravenously or subcutaneously. In addition, commercial insulin is a product made by extracting and purifying from the pancreas of animals such as cows or pigs, and the effect expression time, maximum effect time, and effect duration vary depending on the type. It is necessary to determine the type and dosage of insulin that is most suitable for the patient. Since the optimal time and meal time after the insulin injection are effective, the meal time has to be thoroughly followed.

Oral hypoglycemic agents are used to regulate blood glucose levels in NIDDM patients because they enhance the sensitivity of the insulin receptors in the cells and stimulate the pancreas to promote insulin secretion. However, oral therapies for the treatment of NIDDM have been very limited until recently.

Oral hypoglycemic agents currently used in NIDDM include five groups of compounds: biguanides, thiazolidinediones, sulfonylureas, benzoic acid derivatives, and α-glucose. Α-glucosidase inhibitors.

Sulfonylureas, such as tolbutamide and glyburide, and meglitinide-based drugs block the calcium channels in the beta cells of the pancreas to promote the secretion of engineered insulin Drops blood sugar through. Biguanide-based compounds such as metformin improve blood sugar by inhibiting hepatic angiogenesis and decreasing insulin resistance by enhancing glucose uptake of peripheral cells. Thiazolidinedione-based compounds, such as troglitazone, rosiglitazone and pioglitazone, reduce peripheral insulin resistance to reduce blood sugar. Α-glucosidase inhibitors, such as acarbose and miglitol, inhibit intestinal absorption by preventing the breakdown of disaccharides or polysaccharides in the small intestine mucosa.

These drugs were initially used alone, but in general, since monotherapy has been found to be inadequate, it has been used in the form of complex medications, taking advantage of the complementary effects of different mechanisms of action targeting hyperglycemia. WO 97/17975 and US Pat. No. 5,922,769 disclose the combined use of metformin and sulfonylurea glyburide (glybenclamide), and European Patent Application No. 98401781.4 discloses a formulation comprising metformin and glyburide, US Patent 5,631,224 discloses a combination of metformin and GLP-1 (7-36) amide or GLP-1 (7-37) or fragments thereof. WO 98/57634 discloses a method for treating diabetes using a combination of thiazolidinedione and metformin, and US Pat. No. 5,965,584 discloses a combination of metformin and thiazolidinedione insulin sensitivity enhancers (e.g., pioglitazone). Doing.

However, the sulfonylurea compound cannot be administered to IDDM patients who are insulin dependent, and intensive use of prolonged sulfonylureas increases the risk of hypoglycemia. Female patients can cause side effects such as abnormal fetal birth, abortion and stillbirth, and most of the sulfonylurea compounds should be administered carefully to patients with liver and kidney function impairment due to sulfonylurea metabolism.

Biguanide compounds, such as metformin, do not increase pancreatic insulin secretion but lower blood sugar more effectively than sulfonylureas and have a lower incidence of hypoglycemic shock. However, it can cause nausea, vomiting, diarrhea and rash at the beginning of administration, and can cause serious side effects such as fatal lactic acidosis, which should be used with great care and limited in clinical use in the United States. . In addition, long-term use of oral hypoglycemic agents may cause cardiovascular or gastrointestinal and liver disorders.

Because of these shortcomings and side effects, they are satisfactory in current treatments and at the same time have high safety and safety in terms of side effects, and there are few drugs that can be applied to all diabetics, and thus more effective drugs to treat diabetes, especially NIDDM. Is urgently needed.

The present invention has been attempted to find a natural herbal composition effective for the treatment of such diabetes and thereby complications.

The herbal extract according to the present invention confirmed the efficacy of type II diabetes through animal experiments using the type II diabetes model dividi mouse, and also pancreatic, small intestine, kidney and insulin dependent tissues (muscle, fat, liver, etc.) The effects of insulin, secretion of glucose into the tissues, carbohydrate metabolism and uptake, etc. were extracted from experimental animals, and the mechanism of drug efficacy was investigated by using biochemical and molecular biological techniques at the tissue or cell level.

Seedlings of Cuscuta japonica Choisy, C. australis R. and C. chinensis Lam are known as potting and tonic, and have a resin glycoside, vitamin A. It is also known to contain β-carotene-5, 6-epoxide, taraxanthin, and lutein. Younglimsa, 1998, p. 882-883).

The present inventors found that the Tosa extract exhibits excellent blood sugar lowering action and blood free fatty acid and triglyceride reducing action, and thus shows an excellent effect in the prevention and treatment of diabetes mellitus and its various complications, and by applying a composition comprising the same. Completed.

It is an object of the present invention to provide a pharmaceutical composition and a dietary supplement that have a prophylactic and therapeutic effect on diabetes and diabetes-related diseases.

1 is a view showing the effect on the weight change of the earth and sand extract,

Figure 2 is a diagram showing the blood sugar lowering effect of the Tosa extract,

Figure 3a is a diagram showing the effect of reducing the insulin concentration and insulin resistance of the Tosa extract,

Figure 3b is a diagram showing the effect of promoting insulin secretion in vitro experiments of Tosa extract,

Figure 4 is a diagram showing the effect of preservation of pancreatic beta cells by the extract of Tosa.

Figure 5a is a diagram showing the effect on the blood glucose increase inhibition of the soil water extract and partially purified butanol soluble fraction in oral glucose-load experiment,

FIG. 5B is a diagram showing the AUC obtained during 120 minutes of glucose administration to show the effect of inhibiting the increase in blood glucose of the soil extract and the partially purified butanol soluble fraction in the oral glucose-load experiment.

Figure 6a is a diagram showing the effect on the blood glucose increase inhibition of the soil ethanol extract in oral glucose load experiment.

FIG. 6B is a diagram showing the AUC obtained during 120 minutes of glucose administration to determine the effect of the soil ethanol extract on the blood glucose increase inhibition in the oral glucose-load experiment.

In accordance with the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus and the complications thereof, including the earth and sand extract and a pharmaceutically acceptable carrier, diluent or excipient.

The earth and sand of the present invention is selected from the seeds of the plant of the genus Cuscuta, such as Cuscuta japonica Choisy, C. australis R. and C. chinensis Lam, and preferably C. chinensis Lam) seeds.

The extract includes water, a lower alcohol having 1 to 4 carbon atoms and a mixed solvent thereof, an extract available in a polar solvent, preferably water.

Hereinafter, the present invention will be described in detail.

The earth and sand extract of the present invention is about 1 to 20 times the weight of the seeds of crushed mulberry bird, preferably about 2 to 10 times the volume of water, lower alcohol of 1 to 4 carbon atoms or about 1: 0.1 to 1:10 , Preferably a mixed solvent having a mixing ratio of 1: 0.2 to 1: 3 at an extraction temperature of 20 to 100 ° C, preferably 70 to 90 ° C for about 1 hour to 2 days, preferably 2 hours to 1 day Extraction is repeated once to five times, preferably two to four times, by an extraction method such as hot water extraction, cold sediment extraction, reflux cooling extraction, or ultrasonic extraction, and then the extract is 20 to 100 ° C. with a vacuum rotary concentrator. The residue extracted after concentration under reduced pressure at 40 to 70 ° C. may be dried with a vacuum freeze dryer to obtain earth and sand extract available in water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

In addition, conventional fractionation can also be carried out (Harborne J.B .; Phytochemical methods: A guide to modern techniques of plant analysis. 3rd Ed. P 6-7, 1998). Suspended in water or a lower alcohol extract having 1 to 4 carbon atoms in water, extracted with a solvent such as hexane, dichloromethane, chloroform, ethyl acetate, butanol, which are not mixed with water, insoluble ingredients are removed and diabetes active ingredients are contained. By preparing an effective fraction, it is possible to obtain a partially purified extract which minimizes side effects and expresses efficacy even at a small dose.

The present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus and its complications, which has the effective fraction of the earth and sand extract obtained by the above method and the partial purification effective fraction of the earth and sand extract.

The composition of the present invention comprises 0.1 to 50% by weight of the extract relative to the total weight of the composition.

Complications caused by diabetes include a combination of one or more of chronic hyperglycemia, atherosclerosis, microangiopathy, kidney disease, heart disease, diabetic retinopathy and other ophthalmic diseases. do.

The composition containing the earth and sand extract of the present invention may further comprise a suitable carrier, excipient and diluent according to a conventional method.

Carriers, excipients and diluents that may be included in the compositions of the present invention 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.

The composition comprising the earth and sand extract according to the present invention, in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to a conventional method. Can be formulated and used.

Specifically, when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use may include various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to water and liquid paraffin, which are commonly used to include suspensions, solutions, emulsions, and syrups. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of the extract of the present invention may vary depending on the age, sex, and weight of the patient, but in general, an amount of 0.01 to 1000 mg / kg, preferably 0.1 to 500 mg / kg, may be administered once or several times a day. Can be. The dosage may also be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Tosa extract of the present invention is a drug that can be used with confidence even when taking a long term for the purpose of prevention because there is little toxicity and side effects.

The present invention provides a dietary supplement comprising earthworm extract and food acceptable food additives exhibiting an effect of preventing and improving diabetes and its complications.

The composition containing the earth and sand extract of the present invention can be used in a variety of drugs, foods and beverages for the prevention and improvement of diabetes and its complications. Foods to which the extract of the present invention may be added include various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

At this time, the amount of the extract in the food or beverage, in general, may be added to 0.01 to 15% by weight of the total food weight in the case of the health food composition of the present invention, 0.02 to 10g, based on 100ml for the health drink composition It may be added at a ratio of 0.3 to 1g.

The health beverage composition of the present invention has no special limitation except for containing the earth and sand extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. . Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

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 adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

As a result of the preliminary experiments, the composition of the present invention was observed to have an excellent effect on improving hyperlipidemia such as lowering free fatty acids, triglycerides, cholesterol, etc. in the blood, suggesting that insulin resistance of type 2 diabetes can be improved. Thus, three weeks of oral administration of Tosa extract using type 2 diabetic animal model (db / db mouse), it was confirmed that the excellent hypoglycemic activity and hyperlipidemic effect.

The present invention is described in more detail based on the following examples, although the present invention is not limited thereto.

Example 1 Preparation of Earthenware Water Extract

200 g of crushed sea cucumber seeds (purchased at Asan-Dang Herbal Medicine Herb) were put in 2 liters of water and extracted with boiling water twice for 2 hours with a reflux condenser. The filtrate obtained by filtration with four layers of gauze was concentrated under reduced pressure and lyophilized to obtain 91 g of powdery earthworm extract.

Example 2 Preparation of Butanol Fraction of Water Extract from Earth and Sand

After dissolving 40.5 g of the soil extract obtained in Example 1 in 160 ml of water, shaking twice with an equal amount of saturated butanol to obtain a butanol layer, and then concentrated under reduced pressure to obtain a partially purified 1.7 g butanol soluble extract. .

Example 3. Preparation of Tosa Ethanol Extract

1 g of ethanol is added to 100 g of crushed sea cucumber seed (purchased at Asan-Dang Herbal Medicine of Gyeongsangnam-do, Seoul), and refluxed with a reflux condenser and heated for 2 hours. After drying the residue, the reflux condenser is cooled with 1 liter of ethanol. Sweet and warm extracted twice for 2 hours. The ethanol extract was filtered and the filtrate was concentrated under reduced pressure to give 1.96 g of ethanol extract.

Experimental Example 1. Observation test of diabetes-related indicators in body weight and blood and urine

In this experiment, an 11-week-old male Divi mouse (Male db / db mouse, 50 g, Korea Research Institute of Bioscience and Biotechnology), a type 2 diabetic animal model, was purchased and maintained at a constant temperature (25 ° C) and humidity (50%). Adapted weekly and used in experiment

The mice were divided into four groups, respectively, into the experimental group (sample treated group) and the control group (sample untreated group), and the experimental group was orally administered to the soil extract of Example 1 by dissolving water in water at 500 mg / kg body weight for 3 weeks every day. The control group was fed tap water.

Changes in body weight were measured weekly, and changes in urine glucose and drinking water intake were measured daily. In addition, blood obtained by orbital blood collection with a heparinized capillary was centrifuged to obtain plasma, and glucose and triglyceride concentrations, free fatty acid concentrations, and insulin concentrations were measured therefrom. Blood collection was always performed at 3 pm.

Statistical processing method

All experimental results were expressed as mean ± standard error, and statistical processing was performed by the Student-T test to determine the significance based on P <0.05.

In the following tables and figures, * P, ** P and *** P are * P <0.05, ** P <0.01, *** P <0.001 for the untreated group.

1. Weight change

As shown in FIG. 1, body weight showed a significant decrease compared to the control group from day 12 of administration.

2. Changes in urine glucose and drinking water intake

As shown in Table 1, the urinary glucose and drinking water intake after 3 weeks of administration of the Tosa extract of Example 1 also showed a significant decrease, and it was confirmed that the main symptoms of diabetic and improved effects on the following.

Control Experimental group Urine glucose (mg / mouse / day) 194.5 ± 30.1 1.0 ± 0.5 *** Drinking Water Intake (ml / mouse / day) 7.2 ± 0.2 5.5 ± 0.5 *

3. Blood Sugar Change

Once weekly changes in blood glucose levels of the control and experimental groups were measured.

As shown in Figure 2, the blood glucose of the experimental group showed a significant drop effect at 1 and 3 weeks compared to the control group.

4. Geological Change

In order to examine the effects of Tosa extract on blood cholesterol, triglycerides and free fatty acids, the experiment was conducted as follows.

Plasma Total Cholesterol was measured as follows; Since cholesterol in plasma exists in the form of cholesteryl ester and free cholesterol, in order to quantify both forms, cholesteryl ester was hydrolyzed and decomposed into fatty acids and free cholesterol. The converted free cholesterol and free cholesterol in plasma are converted to Δ ⁴ -cholestenone by cholesterol oxidase and then peroxidase and substrates H ₂ O ₂ , phenol and 4-amino-antipyrine (4- It was reacted with amino-antipyrine) to develop a red color, and absorbance was measured at 500 nm and quantified by comparison with cholesterol standard solution.

Triglyceride in plasma was measured by the following method; After triglyceride in plasma was decomposed into glycerol and fatty acid using lipoprotein lipase, the resulting glycerol was converted to L-α-phosphoglycerol by reaction with glycerol kinase and ATP. Here glycerol in-phospho-oxidase (glycerol-phospho-oxidase; GPO ) the reaction of and to H ₂ O ₂ is generated, H ₂ O ₂ gape hydroxy is measured at 550 nm and then presented azepin reacting with color intensity It was quantified in comparison with the glycerol standard solution (measurement kit; Cleantech TG Asan Pharmaceutical Co., Ltd.).

As a result, as shown in Table 2, the experimental group showed a significant improvement in blood cholesterol concentration, triglyceride concentration and free fatty acid concentration in comparison with the control group.

Control Experimental group Plasma total cholesterol (mg / dl) 141.7 ± 7.1 77.5 ± 1.2 ** Triglycerides (mg / dl) 176.7 ± 4.3 116.7 ± 8.2 * Free fatty acid (uEq / l) 2258.9 ± 90.9 11543.8 ± 34.1 **

5. Changes in Insulin Levels in Blood

In vivo experiments and in vitro experiments were conducted to observe the effect of Tosa extract on insulin. In vivo experiments were performed using ELISA kit (SHIBAYAGI, Japan) to measure the insulin level of dividing mice administered with Tosa extract of Example 1 for 3 weeks, and in vitro experiments were performed on HIT-T15 cell line Tosa extract of 1 was treated for 1 hour at a concentration of 1 mg / ㎖ and the amount of insulin secreted using the RIA kit (DPC, USA) was measured. As shown in Figure 3a and Figure 3b, the experimental group administered the tosa extract of Example 1 showed a significant decrease in blood insulin concentration compared to the control group. This suggests that insulin concentration is reduced by improving insulin resistance characteristic in DI mice. In addition, in vitro experiments showed the same pattern of insulin secretion as treated with glucose at the same concentration as the extract.

Experimental Example 2. Beta cell preservation effect experiment

In this experiment, in order to test the effect of the composition on the beta cells (β-cell) of the pancreas was isolated from the Langerhans islet of the pancreas from the DI mice administered the sample.

The pancreas of mice (experimental) and non-administered mice (control) for 3 weeks were separated and embedded in paraffin and stained with tissues by immunohistochemistry to measure the infiltration of immune cells (insulin). Monoclonal antibody, Neomarkers, USA).

As a result, as shown in FIG. 4, it was confirmed that the insulin content in the beta cells was maintained higher in the experimental group having lower blood insulin concentration than the control group having higher blood insulin concentration. It was observed that insulin in the beta cells of the control group was reduced and depleted. This fact suggests that hyperinsulinemia and damage to beta cells are due to insulin resistance. On the other hand, in the experimental group, it is observed that insulin is conserved in the beta cells at the same time that the insulin in the blood is reduced, indicating that the beta cells are preserved.

Experimental Example 3. Oral Glucose Tolerance Test

To test the glucose tolerance of Tosa extract, the oral glucose loading test was performed after the administration of the sample to compare the hypoglycemic effect between the experimental group and the control group.

1. OGTT Experiments on Soil Extracts and Soluble Fraction of Butanol

Six-week-old ICR mice were divided into five groups, and the control group was administered saline at a dose of 10 ml / kg, and the experimental group was excised from the soil water extract of Example 1 (CSW) and partially purified butanol soluble extract of Example 2 (CSW- 1) was orally administered at doses of 405 mg / kg and 17 mg / kg, respectively, equivalent to 1 g / kg of earth and sand. After 30 minutes, glucose was orally administered at a concentration of 1.5 g / kg, and blood was collected from the orbital vein of the mouse at 30, 60, 90, and 120 minutes to measure blood glucose, and the increase from the blood glucose level before glucose administration. Indicated.

As shown in Fig. 5a, the experimental group administered the soil extract (CSW) at 405 mg / kg dose and the partially purified butanol soluble extract (CSW-1) at 17 mg / kg dose from 30 minutes of glucose administration. Showed a significant effect of inhibiting the blood glucose increase compared to the control group. Figure 5b shows the area under the curve of the oral glucose loading test results for 120 minutes showed a significant blood glucose inhibitory effect. In particular, the partially purified butanol soluble extract fraction (CSW-1) shows a similar effect as water extract even with a significantly smaller dose, which has advantages such as a dose reduction in formulation design.

2. OGTT Experiment on Tosa Ethanol Extract

Tosa ethanol extract (CSE) obtained in Example 3 was orally administered at a dose of 39 mg / kg corresponding to 1 g / kg of Tosa medicinal herbs, and the OGTT experiment was performed in the same manner as in Experimental Example 3-1. As a result, as shown in Figures 6a and 6b it showed a significant increase in blood glucose.

Experimental Example 4. Acute Toxicity Preliminary Study

Example of the present invention by dividing 25 ± 5 g of ICR mice (Korean experimental animals) and 235 ± 10 g of SPF, Sprague Dawley (Biogenomics) rats into three groups Tosa extract of 1 was intraperitoneally administered at doses of 20 g / kg, 10 g / kg and 1 g / kg, respectively, and observed for 24 hours.

As a result, no deaths were observed in all three groups, and no significant symptoms were found in weight gain and feed intake. Therefore, it was confirmed that the extract of the present invention is a safe drug.

The preparation example of the pharmaceutical composition comprising the earth and sand extract of the present invention will be described, but the present invention is not intended to limit it, but is intended to explain in detail only.

Formulation Example 1 Preparation of Powder

Example 1 Dry Powder ............... 300 mg

Lactose ......................................... 100 mg

Talc ........................................ 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Example 2 Dry Powder ............... 50 mg

Corn Starch ... 100 mg

Lactose ............ 100 mg

Magnesium Stearate ............... 2 mg

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Example 3 Dry Powder ............... 50 mg

Corn Starch ... 100 mg

Lactose ............ 100 mg

Magnesium Stearate ............... 2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Example 1 Dry Powder ............... 300 mg

Sterile distilled water for injection ..................

pH adjuster .....

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Example 1 Dry Powder ......................................... 1 g

Isomerized sugar ......................................... 10 g

Mannitol ........................ 5 g

Purified water .........................

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

Example 1 Dry Powder ............... 1000 mg

Vitamin Blend .........................

Vitamin A Acetate ......... 70 μg

Vitamin E ............................. 1.0 mg

Vitamin B ₁ ........................................ 0.13 mg

Vitamin B ₂ ............. 0.15 mg

Vitamin B ₆ ............................. 0.5 mg

Vitamin B ₁₂ ......................... 0.2 μg

Vitamin C ........................ 10 mg

Biotin ................................... 10 μg

Nicotinic acid amide ......... 1.7 mg

Folic acid .................................... 50 μg

Calcium Pantothenate ......................................... 0.5 mg

Inorganic Mixtures ...

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium Carbonate ............... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dicalcium Phosphate Diluent ... 55

Potassium Citrate ............... 90 mg

Calcium Carbonate ... 100 mg

Magnesium Chloride ............... 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

Dry powder of Example 1 ...................................... 1000 mg

Citric Acid ......................................... 1000 mg

Oligosaccharide ......................................... 100 g

Plum concentrate ........................... 2 g

Taurine ......................................... 1 g

Purified water is added to the whole .............. 900 ml

After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Tosa extract of the present invention exhibits excellent hypoglycemic action and hyperlipidemic action, the composition comprising the same can be used as a pharmaceutical and health functional food useful for the prevention and treatment of diabetes mellitus and its various complications.

Claims (10)

A pharmaceutical composition for treating diabetes mellitus and thereby complications, comprising a tosa extract as an active ingredient, and containing a pharmaceutically acceptable carrier, diluent or excipient. The pharmaceutical composition of claim 1, wherein the extract is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The pharmaceutical composition according to claim 1, wherein the earthworm is selected from the group consisting of seeds of Cuscuta japonica Choisy, C. australis R. and C. chinensis Lam. The pharmaceutical composition according to claim 1, wherein the earthworm is a seed of C. chinensis Lam. The pharmaceutical composition according to claim 1, wherein the complications caused by diabetes include a combination of one or more of chronic hyperglycemia, atherosclerosis, microangiopathy, kidney disease, heart disease, diabetic retinopathy and other ophthalmic diseases. . The pharmaceutical composition according to claim 1, wherein the composition is formulated in the form of a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol oral formulation, external preparation, suppository or sterile injectable solution. A pharmaceutical composition for treating diabetes mellitus and thereby complications comprising a polar solvent soluble extract fraction of earth and sand as an active ingredient, and containing a pharmaceutically acceptable carrier, diluent or excipient. The pharmaceutical composition of claim 7, wherein the extract is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. A dietary supplement comprising earthworm extract and food-acceptable food supplement additive that has a prophylactic and improvement effect on diabetes and its complications. 10. The dietary supplement of claim 9 in the form of a powder, granule, tablet, capsule or beverage.