WO2012105816A2 - Composition for preventing and treating diabetes and diabetes complications comprising amphicarpaea edgeworthii var. trisperma powder or an extract thereof - Google Patents

Abstract

The present invention relates to a composition for preventing and treating diabetes comprising Amphicarpaea edgeworthii var. trisperma powder or an extract thereof which have a blood glucose lowering effect, and the Amphicarpaea edgeworthii var. trisperma powder or extract thereof of the present invention can be used to advantage in pharmaceutical compositions and health and functional foods for preventing and treating diabetes by suppressing and regulating sudden rises in blood glucose concentration.

Description

Composition for preventing and treating diabetes mellitus and diabetic complications comprising soybean powder or extracts thereof

The present invention relates to a composition for the prevention and treatment of diabetes mellitus and diabetic complications containing a powder ( Aphicarpaea edgeworthii var. Trisperma ) powder or extract thereof as an active ingredient having a hypoglycemic effect.

Diabetes mellitus is a disease in which hyperglycemia occurs because cells in the body cannot use sugar due to decreased insulin secretion or reduced insulin function. In diabetes mellitus, hormone imbalance, including insulin, is a characteristic of hyperglycemia due to abnormal physiological metabolic control functions such as carbohydrate-containing protein, lipid and electrolyte metabolism, and if such hyperglycemia persists, blood circulation disorders, retinal damage, nerve cell damage, It causes renal insufficiency and vascular complications and serious chronic complications.

In particular, the incidence of cardiovascular diseases such as atherosclerosis, cerebral infarction, cerebral thrombosis, and myocardial infarction is higher in diabetic patients than normal patients (Fuller, J.H., Lancet, 1, pp1373-1376, 1980). Diabetes patients have a high risk of death from coronary artery disease or cerebrovascular disease, which are often caused by hypertension, hyperlipidemia, obesity, etc. (Hu-Beom Bum. The Korean Nutrition Society. Abstract, pp. 15-18, 1984). It has been reported that 67% of type 2 diabetics have one or more types of lipid metabolism (Harris, M.I. Diabetes Care, 23, pp754-758, 2000). Type 2 diabetics have increased lipid metabolism with increased triglyceride and cholesterol levels and decreased HDL-cholesterol (Goldberg, RB Diabetes Care, 4, pp561-572, 1981). (Reaven, JW Am. J. Med., 83, pp 31-40, 1987).

Diabetes is defined as a metabolic disorder caused by insulin secretion and a lack of action of insulin secreted by pancreatic cells and involves excessive production of glucose, breakdown of body fat and waste of protein, and abnormal glands of glucagon, resulting in metabolic disruption. Causes (Abrams, JJ, Ginsberg, H, et al., Diabetes, 31, pp 903-910, 1982).

Diabetes mellitus is characterized by two types.Type 1 diabetes mellitus is caused by a deficiency of insulin, a glucose-regulating hormone in the blood, mainly 10-20. It is called juvenile diabetes because it occurs in younger generations. Type 2 diabetes mellitus occurs mainly after the age of 40 and accounts for most of the diabetic patients in Korea. Unlike type 1, it is called adult-type diabetes and the cause of the disease is not clear yet, but it is known to be caused by genetic factors and environmental factors. As a etiology of type 2 diabetes, both insulin secretion in pancreatic beta cells and defects in insulin action (insulin resistance) in target cells are observed.

The most important goal in the treatment of diabetes is to control blood glucose levels as close to normal as possible. After fasting blood sugar control along with fasting blood glucose is important for improving diabetes and preventing and treating complications. There is exercise therapy.

Oral hypoglycemic agents currently used in patients with type 1 and type 2 diabetes include alpha-glucosidase inhibitors, sulfonylureas, and biguanides. Sidase inhibitors have a therapeutic effect on diabetes by delaying the digestion and absorption of carbohydrates in the ingested diet, thereby reducing the elevation of postprandial blood sugar and blood insulin. Alpha-glucosidase inhibitors promote the secretion of glucagon-like peptide-1, which promotes insulin secretion and suppresses glucagon secretion, without causing hyperinsulinemia or hypoglycemia. Have advantages (Mooradian, AD, Thurman, JE et al., Drugs, 57, pp 19-29, 1999; Baron, AD et al., Diabetes Research and Clinical Practice, 40, ppS54-S55, 1998).

Acarbose, Boglibose, and Miglitol are currently used in the clinic, but long-term use of alpha-glycosidase inhibitors may cause abdominal bloating, vomiting, and diarrhea in some patients. Side effects may be indicated and their use may be limited (Hanefeld, M. et al., Journal of Diabetes and its Complications, 12, pp228-237, 1998).

Bird bean ( Amphicarpaea edgeworthii var. Trisperma ) is a year-old herb belonging to the Zingiberaceae family, a vine-like plant, 1-2m long, with spread hairs facing down. In Korea, it grows wild in wild fields all over the country and is distributed in China, Japan, and India. New beans are also named " Amphicarpaea bracteata subsp.edgeworthii (Benth.) H. Ohashi", " Amphicarpaea edgeworthii var. Japonica ". Leaves are alternate, long petiole, 3 leaflets, ovate, blossom in June-September, flowers are purple. Fruits mature in August. Fruits are stalk, flat oval, with hairs along circumference, with 3 seeds. 10 is a picture of the new beans distributed in Korea can be confirmed the leaves and flowers, Figure 11 is a picture of the seed. In oriental medicine, the root is used as a medicine called Yang Hyung-Doo, which is known to be effective in limb pain. Ingredients include essential oils of seeds, the main components of which are Borneol, Bornyl-acetate, Linalool, Nerolidol, etc. Literature Review on the Form and Form, Korean Journal of Herbology, p96-105, 1986). However, none of the above documents discloses or teaches the medical effects of new beans on diabetes and diabetic complications.

[references]

[1] Fuller, J.H., Lancet, 1, pp1373-1376, 1980

[Reference 2] Hur Bap. Korean Nutrition Society. Abstract paper collection. pp 15-18, 1984

[Reference 3] Harris, M.I. Diabetes Care, 23, pp 754-758, 2000

[4] Goldberg, R.B. Diabetes Care, 4, pp 561-572, 1981

[Reference 5] Reaven, J.W. Am. J. Med., 83, pp 31-40, 1987

Abrams, J.J., Ginsberg, H, et al., Diabetes, 31, pp 903-910, 1982

7 Mooradian, A.D., Thurman, J.E. et al., Drugs, 57, pp 19-29, 1999;

Document 8 Baron, A.D. et al., Diabetes Research and Clinical Practice, 40, ppS54-S55, 1998

[9] Hanefeld, M. et al., Journal of Diabetes and its Complications, 12, pp228-237, 1998

[10] Lee Choong-seop et al., Korean Journal of Herbology, p96-105, 1986

[Reference 11] Am. J. Endocrinol. Metab. 288: E510-E518 (2005)

Document 12; Diabetologia 49: 1647-1655 (2006)

13 J. Ethnopharmacol. 103: 491-495 (2006)

14 J. Med. Chem. 43: 3487-3494 (2003)

Metabolism 50: 1049-1053 (2001).

16 Metabolism 53: 488-499 (2004)

The present invention is to provide a novel substance excellent in postprandial blood sugar suppressing action without any side effects on the living body. To this end, in the present invention, the pharmacological effect of the new soybean powder or its extracts was determined through the blood glucose measurement test using the db / db mouse model, the triglyceride in the serum, the total cholesterol, the blood BUN, the ALT and the AST measurement test. As a result, the present invention was completed by confirming that there is an excellent therapeutic effect on diabetes, lipid metabolism improvement, and complications caused by diabetes.

An object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of diabetes mellitus or diabetic complications containing a soybean powder or extract thereof as an active ingredient having an excellent hypoglycemic effect.

It is also an object of the present invention to provide a dietary supplement for the prevention and improvement of diabetes mellitus or diabetic complications containing a soybean powder or its extract as an active ingredient having an excellent hypoglycemic effect.

In the present invention,

There is provided a pharmaceutical composition for the prevention and treatment of diabetes mellitus or diabetic complications containing a powder ( Aphicarpaea edgeworthii var. Trisperma ) or extract thereof as an active ingredient. The extract may be extracted with water, an organic solvent or a mixed solvent thereof. The diabetic complications include arteriosclerosis, cerebral infarction, cerebral thrombosis, myocardial infarction, hypertension, hyperlipidemia, obesity, and the like.

In the present invention,

Provided is a dietary supplement for the prevention and improvement of diabetes mellitus or diabetic complications, which contains a bird bean ( Amphicarpaea edgeworthii var. Trisperma ) powder or an extract thereof as an active ingredient. The health functional food of the present invention may have any form of tablets, capsules, powders, granules, liquids, and pills.

In the present invention,

Provided is a dietary supplement comprising Amphicarpaea edgeworthii var.trisperma powder or extracts thereof which shows the effect of improving blood sugar, improving lipid metabolism and preventing and treating diabetes complications. The food includes food, drink, gum, tea, vitamin complex, or health functional food.

As described above, in the present invention, the blood glucose measurement test using the db / db mouse model, the triglyceride in serum, total cholesterol, total cholesterol, blood BUN, ALT and AST measurement test using a db / db mouse model, diabetes, lipid It was confirmed that there is an excellent therapeutic effect and an excellent hypoglycemic effect on metabolic improvement and complications caused by diabetes. The composition of the present invention using a soybean powder or an extract thereof as an active ingredient may be usefully used as a pharmaceutical composition and health functional food for the prevention and treatment of diabetes or diabetes complications.

1 shows blood glucose in diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 6 weeks) treated or treated with banaba extract, ABSE, B-ABSE (n = 7), respectively. (Figures are mean ± SEM and numbers with different indications differ from one another at specific time points; statistically significant compared to negative control by T test method) * p <0.05, *** p <0.001)).

FIG. 2 shows blood glucose in diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with Banaba extract, ABSE, B-ABSE (n = 7), respectively. (Figures are mean ± SEM and numbers with different indications differ from one another at specific time points; statistically significant compared to negative control by T test method) * p <0.01)).

3 shows blood neutrality in diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with Banaba extract, ABSE, B-ABSE (n = 7), respectively. Figures showing triglyceride levels (values are mean ± SEM and values with different indications differ from one another at specific time points; statistically compared with negative control by T test method (* P <0.01)).

FIG. 4 shows blood blood levels in diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with banaba extract, ABSE, B-ABSE (n = 7), respectively. urea nitrogen level (values are mean ± SEM and values with different indications differ from one another at specific time points; statistically significant compared to negative control by T test method (* P <0.01)).

FIG. 5 shows blood total in diabetic mice (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with Banaba extract, ABSE, B-ABSE (n = 7), respectively. Cholesterol level (values are mean ± SEM and values with different indications are different from each other at each specific time point; statistically significant compared to negative control by T test method) (* p <0.01));

6 shows blood AST in diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with Banaba extract, ABSE, B-ABSE (n = 7), respectively. And ALT levels (values are mean ± SEM and values with different indications differ from one another at specific time points; statistically significant compared to negative control by T test method; (* P <0.01).

7 is a diagram showing the effect on blood AST and ALT levels in db / db mice (Makoto T. et. Al., Pulmonary exposure to diesel exhaust particles enhances fatty change of the liver in obese diabetic mice.INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 19: 17-22, 2007).

FIG. 8 shows average body weight in diabetic rats treated with or treated with banaba extract, ABSE, B-ABSE (n = 7), respectively (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks). Are degrees representing each,

9 shows the dietary intake of diabetic rats (BKS.Cg-m ^{+ / +} Lepr ^{db / J} diabetic mice; 9 weeks) treated or treated with Banaba extract, ABSE, B-ABSE (n = 7), respectively. Are the figures respectively.

10 is a picture of a new bean growing in Korea.

11 is a picture of the new soybean seed growing in Korea.

The composition of the present invention contains a soybean ( Amphicarpaea edgeworthii var. Trisperma ) powder or an extract thereof as an active ingredient.

The soybeans defined in the present invention are plants named by the scientific name " Amphicarpaea edgeworthii var. Trisperma ", or " Amphicarpaea bracteata subsp.edgeworthii (Benth.) H. Ohashi" or " Amphicarpaea edgeworthii var. Japonica ". The soybeans are edible, flat oval, with hairs along the perimeter, with three seeds inside. In the present invention, "soybean" or "soybean seed" means a seed contained in the soybean fruit.

The soybean powder includes both dry powder or powdered powder of soybean seed. The extract is a soybean powder of water, an organic solvent or a mixed solvent extract thereof. More preferably, the extract is an extract extracted with water, an organic solvent or a mixed solvent thereof. The diabetic complications include arteriosclerosis, cerebral infarction, cerebral thrombosis, myocardial infarction, hypertension, hyperlipidemia, obesity, and the like, and particularly include hypertension, hyperlipidemia or obesity.

The soybean powder of the present invention and its extract will be described in detail.

Commercially available dried soybeans can be ground in a grinder to obtain the soybean powder of the present invention in a dry state. In addition, the soybean powder may be increased at 20 to 120 ° C. for about 1 to 72 hours, and dried according to a conventional drying method in the art to obtain the new bean powder of the present invention in the form of a powder. In addition, the soybean powder or the powdered soybean dry powder is lyophilized and pulverized, and then a volume of water or an organic solvent of about 2 to 15 times, preferably about 5 to 10 times the weight of the sample, such as methanol, ethanol, The extract of the present invention can be obtained by extraction with a polar solvent of C ₁ to C ₄ lower alcohols such as butanol, or a mixed solvent thereof. Extraction may be preferably performed using an extraction method such as hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction at 20 to 120 ℃ for about 1 to 72 hours with water, preferably after extraction under reduced pressure filtration and concentration The extract of the present invention can be obtained.

In addition, conventional fractionation processes can also be carried out (Harborne J. B. Phytochemical methods: A guide to modern techniques of plant analysis, 3rd Ed., Pp 6-7, 1998).

In the present invention, the soybean powder or extract thereof of the present invention obtained by the method as described above through the blood glucose measurement experiment using the db / db mouse model, triglyceride in serum, total cholesterol, blood BUN, ALT and AST measurement through The therapeutic effect was confirmed. As a result, it was confirmed that there is an excellent therapeutic effect and excellent hypoglycemic effect on diabetes, lipid metabolism improvement and complications caused by diabetes.

In addition, the new soybean has been used for a long time as an edible or herbal medicine extracts of the present invention extracted therefrom also have no problems such as toxicity and side effects.

The pharmaceutical composition for preventing or treating diabetes mellitus or diabetic complications containing the soybean extract of the present invention comprises the extract in an amount of 0.1 to 50% by weight based on the total weight of the composition.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds, as well as in any suitable collection.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract 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. In the case of formulation, it is formulated using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants which are commonly used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid form preparations may contain at least one excipient such as starch, calcium carbonate, sucrose ( Sucrose) or lactose (Lactose), gelatin, etc. are mixed and formulated. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. 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, Whitepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 10000 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

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

Definitions of the terms of the excipient, binder, disintegrant, lubricant, copulation agent, flavoring agent, etc. of the present invention are known in the art (Korean Pharmacopoeia, Moon Sungsa, Korea Pharmaceutical University Association, 5th edition, p33-48 , 1989), and other meanings that include the same or similar functions and effects.

In addition, the present invention provides a health functional food for preventing or treating diabetes or diabetic complications, which contains a powder of the soybean (Aphicarpaea bracteata subsp.edgeworthii (Benth.) H.Ohashi) having a hypoglycemic effect or an extract thereof as an active ingredient. .

"Health functional food" as defined in the present invention means a food manufactured and processed using raw materials or ingredients having a useful function to the human body, and is substantially a health functional food according to the Act No.6727 of the Health Functional Food Act. It is the same meaning as the definition of. In the present invention, the term "functional" means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action on the structure and function of the human body. The dietary supplement for preventing diabetes of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition. In addition, it is possible to manufacture and process as a health functional food in the form of tablets, capsules, powders, granules, liquids, pills for the purpose of preventing diabetes.

In addition, the present invention provides a dietary supplement added to the food or drink for the purpose of improving the blood sugar, extracts thereof soybean sugar improvement, lipid metabolism and prevention and treatment of diabetic complications. Examples of the food to which the soybean powder or extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods. In this case, the amount of the extract in the food or beverage may be added to 0.01 to 100% by weight of the total food weight, the health beverage composition may be added to about 0.02 to 50g based on 100ml.

The health functional beverage composition of the present invention is not particularly limited in the other components, except for containing the above-mentioned extract as an essential ingredient in the indicated ratio, and contains various flavors or natural carbohydrates, etc. as additional ingredients, like ordinary drinks. can do. 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 conventional sugars such as polysaccharides such as dextrin, cyclodextrin, 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 proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the extract 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. In addition, the sample of the present invention may contain natural fruit juice and fruit flesh for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the sample of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples, Reference Examples and Experimental Examples. However, the following Examples, Reference Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples, Reference Examples and Experimental Examples.

Example 1 Preparation of New Bean Powder and Extract

1-1. Soybean powder and cooked powder

Saekong (Amphicarpaea bracteata subsp.edgeworthii (Benth.) Seeds of the present invention was used as a pulverized and dried from Gyeongbuk Mungyeong woojidong mixer (hereinafter, "ABSE" referred to), 250 ml of the seed to saekong ABSE 500g dry matter after water was added and boiled for 5 minutes at 95-100 ℃, and to obtain a powder (hereinafter referred to as, "B-ABSE") 720g obtained by crushing with a mixer was used for the experiment.

1-2. New Bean Powder and Boiled Powder Extract

150 g each of the soybean gray powder and the increased soybean gray powder obtained in 1-1 were added with 13 l of 70% methanol for 24 hours at 40 ° C., and the filtrate was collected under a reduced pressure (Watman, USA), and the filtrates were vacuum-rotated. Concentrating under reduced pressure at 37 ° C. with a concentrator yielded 19.8 g and 29.6 g of soybean gray powder extract (hereinafter referred to as “ ABS1” ) and the increased soybean gray powder extract (hereinafter referred to as “ B-ABSE1” ), respectively. It was used for the experiment while storing at 20 ℃.

Reference Example 1. Preparation of Type 2 Diabetes Animal Model and Sample

1-1. Test substance

Samples of the above Example: Administration Method: Diluted (suspended) in physiological saline was administered orally twice a day (10 am, 6 pm).

1-2. Positive control substance

As a positive control, Banaba leaf extract (Banaba leaf extract , purchased from Wellis Banaba Co., Ltd., Brown tablet) was made into a powder, and then diluted or suspended in physiological saline and administered orally twice (10 AM and 6 PM). .

1-3. Based on preliminary animal experiment data

Six-week-old male db / db mice (26g, Taconic Farms, Inc.) were used to classify three groups of seven for each antidiabetic group, followed by temperature 22 ± 2 ° C, humidity 55 ± 10%, and 12 hours. Proper hypoglycemic efficacy is confirmed for each sample by measuring body weight and blood glucose at 0, 3, 6, 9, 12, and 15 days after sample administration while maintaining in a SPF (specific pathogen free) environment controlled by intervals of contrast. The concentration was chosen.

1-4. Evaluation of Efficacy of Anti-Natural Diabetic Drugs

A 6-week-old male db / db mouse with obesity and symptoms of diabetes artificially induced genetic mutations to block leptin feedback signaling and grow, resulting in type 2 diabetes with overweight. Drugs were orally administered to this mouse model, and the same amount of saline was administered to the control group. The experimental group db / db mice were randomly assigned to each of the seven dogs, divided into four groups, one group was a vehicle (saline), the positive control group 100mg / kg, and the experimental group prepared in the above Samples of ABSE 2g / kg and B-ABSE 2g / kg were administered orally 0.4ml at 10 AM and 18 PM, respectively. The breeding conditions of each group were the same as those of the preliminary animal experiments. Body weight and blood glucose at 0, 7, 14, and 21 days were maintained while maintaining in an SPF environment controlled by a temperature of 22 ± 2 ° C, 55 ± 10% humidity, and 12 hours of contrast. Changes were performed using a handheld blood glucose meter (OneTouch ^™ , Johnson & Johnson, USA) and averaged for each experimental group.

1-5. Blood analysis Autopsy and pathology

As described above, three samples were administered to db / db mice for 15 days, blood was collected from animals of each group, centrifuged at 3,000 rpm for 10 minutes, and then the supernatant plasma was separated from insulin, AST, ALT, Analyzes of total cholesterol (TC), triglyceride (TG), blood urea nitrogen (BUN), and glucose were performed to determine the difference between control and blood biochemical markers related to sugar lipids and other organ toxicity in the blood. Significance analysis was performed using student t-test.

In addition, the liver, kidney, heart, pancreas, and lungs are excised and stored in 10% neutral buffered formalin and part of the organs are stored in RNASol ^B to perform histopathological observations on major organs.

1-6. Statistical processing

Results from various experiments were recorded as mean ± standard error, and significance test was determined using Student's T-test method.

Experimental Example 1. Antidiabetic effect in BKS.Cg-m ^{+ / +} Lepr ^{db / J} mice, type 2 diabetes animal model

In order to confirm the hypoglycemic effect of the samples of the above example, the experiments were performed by modifying the method disclosed in the literature [Am J Physiol Endocrinol Metab. 288: E510-E518 (2005); Diabetologia 49: 1647-1655 (2006); J. Ethnopharmacol. 103: 491-495 (2006); J. Med. Chem. 43: 3487-3494 (2003); Metabolism 50: 1049-1053 (2001); Nutrition & Metabolism 53: 488-499 (2004).

1-1. Effect on blood sugar change

In the 6-week-old db / db mouse model, 100 mg / kg of positive control Banaba extract, 2 g / kg of ABSE, and 2 g / kg of B-ABSE were administered orally twice a day for 3 weeks in the morning and afternoon. Blood glucose was measured by collecting blood from the tail of the mouse once a week using a portable blood glucose meter, and the diet of all mice was removed the day before the measurement (about 12 hours).

As shown in FIG. 1, the blood glucose of the control group (NC) was increased by more than 70% compared to NC from 342.1 mg / dL at 6 weeks to 582.1 mg / dL at 9 weeks. ABSE was the most suppressed blood glucose change from NC at 6 to 9 weeks of age. At 9 weeks of age, the ABSE-administered group showed 360.5 mg / kg of 37.9% blood glucose reduction compared to NC (p <0.001). The positive control group, banaba extract and B-ABSE, showed 26.2% and 26.4% reductions in blood glucose, respectively (p <0.001).

The 6-week-old db / db mouse model was orally administered with positive control Banaba extract 100 mg / kg, experimental ABSE 2 g / kg, and B-ABSE 2 g / kg twice daily in the morning and afternoon for three weeks. As a result, as shown in FIG. 1, the blood sugar of the control group (NC) was increased by more than 70% compared to NC from 342.1 mg / dL at 6 weeks to 582.1 mg / dL at 9 weeks. ABSE was the most suppressed blood glucose change from NC at 6 to 9 weeks of age. At 9 weeks of age, the ABSE-administered group showed 360.5 mg / kg of 37.9% blood glucose reduction compared to NC (p <0.001). After the end of the experiment, the diet was removed on the day before, blood was drawn on an empty stomach (approximately 12 hours), serum was separated, and the glucose level of NC was 641 mg / dL for glucose change in serum using a serum autometer. Among the natural products, the ABSE-treated group showed 427.0 mg / dL of 33.4% significant decrease compared to the NC group (p <0.01).

These results can be confirmed that the hypoglycemic effect of ABSE as a result almost equivalent to the results of blood glucose changes in the tail vein of FIG.

1-2. Measurement of glucose change in serum

After 9 weeks of age, the diet was removed the day before the end of the experiment, anesthetized with ethyl ether in a fasting state (about 12 hours), blood was collected from the heart with a 3ml syringe, left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 10 minutes. Serum was separated and the glucose (glucose) change in serum was measured with a serum autometer, and the results are shown in FIG. 2. The glucose level of NC was 641 mg / dL, and the ABSE-treated group was 427.0 mg / dL, showing a 33.4% significant reduction compared to NC group (p <0.01).

* As a result of this experiment, the anti-diabetic effect of ABSE was observed in the result almost consistent with the results of blood glucose change in the tail vein. Banaba extract, a positive control, was 463 mg / dL, which showed a statistically significant decrease of 27.7% compared to the NC group (p <0.01). However, the B-ABSE group was 551 mg / dL, slightly lower than the NC group by 14%, and there was no statistical significance.

Experimental Example 2. Effect on lipid metabolism and diabetic complications in BKS.Cg-m ^{+ / +} Lepr ^{db / J} mice

In order to confirm the effects of lipid metabolism and diabetic complications of the samples of the examples, the experiments were carried out by modifying the method disclosed in the literature [Am J Physiol Endocrinol Metab. 288: E510-E518 (2005); Diabetologia 49: 1647-1655 (2006); J. Ethnopharmacol. 103: 491-495 (2006); J. Med. Chem. 43: 3487-3494 (2003); Metabolism 50: 1049-1053 (2001); Nutrition & Metabolism 53: 488-499 (2004).

2-1. Measurement of Triglyceride (TG) Changes in Serum

After 9 weeks of age, the diet was removed the day before the end of the experiment, anesthetized with ethyl ether in a fasting state (about 12 hours), blood was collected from the heart with a 3ml syringe, left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 10 minutes. Serum was separated and the serum triglyceride (TG) change was measured with a serum autometer. The results are shown in FIG.

As shown in FIG. 3, the triglyceride (TG) level of NC was 67.0 mg / dL , and the ABSE-treated group was 37.0 mg / dL among the natural-treated groups, which showed a statistically significant reduction of more than 44.7%. (P <0.001). These results were almost consistent with the results of glucose suppression effect in serum, and the blood improvement effect of ABSE was observed. Banaba extract, a positive control, was 50.0 mg / dL, which showed a statistically significant decrease of 25.3% compared to the NC group (p <0.01). However, the B-ABSE group was 55.0 mg / dL, which was slightly reduced to 17.9% compared to the NC group, and there was no statistical significance.

2-2. Measurement of blood urea nitrogen (BUN) changes in serum

After 9 weeks of age, the diet was removed the day before the end of the experiment, anesthetized with ethyl ether in a fasting state (about 12 hours), blood was collected from the heart with a 3ml syringe, left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 10 minutes. Serum was separated and the BUN change in serum was measured with a serum autometer, and the results are shown in FIG. 4.

As shown in FIG. 4, the BUN level of NC was 27.1 mg / dL, and the ABSE-administered group was not significantly different from the 29.4 mg / dL B-ABSE-administered group compared to the 29.8 mg / dL NC group. Banaba extract was 36.0 mg / dL, showing a statistically significant increase of 32.8% compared to the NC group (p <0.05).

2-3. Determination of Total Cholesterol Changes in Serum

After 9 weeks of age, the diet was removed the day before the end of the experiment, anesthetized with ethyl ether in an empty stomach (about 12 hours), the blood was drawn from the heart with a 3ml syringe, left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 10 minutes. Serum was separated and the total amount of cholesterol in the serum was measured by a serum autometer, and the results are shown in FIG. 5.

As shown in FIG. 5, the total cholesterol level of NC was 142.0 mg / dL, and the Banaba extract-treated group was 150.0 mg / dL in the natural product group, 151.0 mg / dL in the ABSE group, and 151.0 mg / d in the B-ABSE group. There was no difference between the experimental groups by dL.

2-4. Measurement of ALT and AST Changes in Serum

After 9 weeks of age, the diet was removed the day before the end of the experiment, anesthetized with ethyl ether in a fasting state (about 12 hours), blood was collected from the heart with a 3ml syringe, left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 10 minutes. Serum was isolated and serum ALT and AST levels were measured with a serometry. The results are shown in FIG.

As shown in FIG. 6, the ALT level was 78.0 U / L in the NC, and the Banaba extract-administered group was 65.5 U / L, the ABSE-administered group was 100.0 U / L, and the B-ABSE-administered group was 108.8 U / L. L and the positive control Banaba extract administration group showed liver function improvement effect compared to NC. The AST level was 189.4 U / L for NC, 90.1 U / L for the barnabas extract group, 171.9 U / L for the ABSE group, and 193.9 U / L for the B-ABSE group. The administration group showed a significant decrease of 51.9% compared to NC, which is close to the normal liver function level of the mice. However, the experimental group ABSE group and B-ABSE group did not differ from NC. In addition, the db / db mouse model is considered to be the reason why the AST level is higher than that of normal mice in the obese / type 2 diabetes model (FIG. 7).

Experimental Example 3. Effect on weight and dietary changes

Six-week-old db / db mice were used in each experimental group, and the change in body weight was measured twice a week. The dietary intake was measured every three days, and the average daily dietary intake of each experimental group (7) was observed. The results are shown in FIGS. 8 and 9.

As a result of the experiment, as shown in Figures 8 and 9, the change in body weight did not show a significant change in body weight in the NC, Banaba extract administration group, ABSE administration group, B-ABSE administration group. However, the daily intake increased significantly after 7 weeks of age, and the dietary intake of ABSE and B-ABSE groups increased by 26.7% and 10.7%, respectively, after 8 weeks. However, the body weight of ABSE and B-ABSE groups did not increase in proportion to the increase in dietary intake.

In the diabetic db / db mice, the soybean powder (ABSE) and boiled soybean (B-ABSE) groups significantly decreased blood glucose and triglyceride levels (p <0.01, p <0.001) compared to the non-administered group. The soybean meal (ABSE) group and boiled soybean (B-ABSE) group showed higher dietary intake, no weight change, and no gross abnormalities were observed. In particular, the group treated with soybean powder (ABSE) showed hypoglycemic effect to alleviate fasting hyperglycemia after fasting.

Experimental Example 4. Acute Toxicity Test

Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats. Two animals of each group were orally administered with the soybean extract of the present invention at a dose of 100 mg / kg. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to visually observe abnormalities in organs and thoracic organs.

As a result of this experiment, there were no clinical symptoms or dead animals in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemical test and autopsy findings. As a result, the extract of the present invention did not show a change in toxicity even in rats up to 100 mg / kg, respectively, and the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance of 100 mg / kg or more.

Hereinafter, the preparation examples of the pharmaceutical composition comprising the extract of the present invention will be described. Such formulations are not intended to limit the present invention, but merely to be described in detail.

Formulation Example 1 Preparation of Powder

ABSE1 Extract 20 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

ABSE1 Extract 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation Example 3 Preparation of Capsule

ABSE1 Extract 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 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

ABSE1 Extract 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

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

ABSE Extract 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, and lemon flavor is added, the above components are mixed, and then, purified water is added to adjust the total amount to 100 ml, and then sterilized by filling into a brown bottle. do.

Claims (7)

1. A pharmaceutical composition for the prevention and treatment of diabetes mellitus or diabetic complications containing Amphicarpaea edgeworthii var. Trisperma powder or extract thereof as an active ingredient.
2. The pharmaceutical composition of claim 1, wherein the extract is extracted with water, an organic solvent or a mixed solvent thereof.
3. The pharmaceutical composition of claim 1, wherein the diabetic complication is arteriosclerosis, cerebral infarction, cerebral thrombosis, myocardial infarction, hypertension, hyperlipidemia, or obesity.
4. A dietary supplement for the prevention and improvement of diabetes mellitus or diabetic complications, which contains Amphicarpaea edgeworthii var.trisperma powder or its extract as an active ingredient.
5. The health functional food according to claim 4, wherein the health functional food has any one form of tablet, capsule, powder, granule, liquid, and pill.
6. A dietary supplement comprising Amphicarpaea edgeworthii var. Trisperma powder or extracts thereof which shows the effect of improving blood sugar, improving lipid metabolism and preventing and treating diabetic complications.
7. The food according to claim 6, wherein the food is food, beverage, gum, tea, vitamin complex, or health functional food.

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