KR100656086B1 - alpha-glucosidase inhibitor - Google Patents

Abstract

In the present invention, jinjin (진) mugwort ethanol and organic solvent extracts were prepared, it was confirmed that the extract has excellent alpha glucosidase inhibitory activity in vitro . In in vivo experiments, oral administration of Injin mugwort extract (400 mg / kg) with maltose (2 g / kg) significantly reduced postprandial blood glucose in rats and diabetic rats compared with maltose. In addition, oral administration of Injin mugwort extract to the prepared plain yogurt significantly reduced postprandial blood sugar increase compared to the control group. Therefore, the extract of Injin mugwort has a very excellent effect that can be used to prevent and / or treat diabetes in humans or animals by suppressing a rapid rise in blood glucose levels after eating.

Mugwort, extract, alpha-glucosidase, hypoglycemic

Description

Phosphorus mugwort extract and composition for inhibiting postprandial hyperglycemia containing the same {alpha-glucosidase inhibitor}

1 is a diagram showing the effect of suppressing postprandial blood sugar in the case of administration of jinseng extract (400mg / kg) with maltose in streptozotocin-induced diabetic rats.

2 is a diagram showing the effect of suppressing postprandial blood sugar after ingesting the extract of Injin mugwort in normal rats.

Figure 3 is a diagram showing the effect of suppressing postprandial blood sugar after injin mugwort yogurt intake in normal rats.

The present invention relates to an extract of Injin mugwort having a hypoglycemic effect. More specifically, the present invention contains an extract obtained by extracting Injin mugwort with alcohol (ethanol) as an active ingredient, and the extract of the active ingredient is alpha-glucosidic in the intestine. The present invention relates to a composition which is useful as a hypoglycemic agent that inhibits glucose absorption by the human body by inhibiting day-glucosidase activity. Diabetes causes abnormal physiological metabolic control functions such as carbohydrates, protein, lipid and electrolyte metabolism due to hormone imbalances such as insulin, glucagon and glucocorti-coid. It is characterized by high blood sugar and diabetes, and if the diabetes persists, it causes serious chronic complications including blood circulation disorders. Diabetes destroys pancreas β-cells according to the mechanisms that occur largely. Insulin-dependent diabetes mellitus is classified as insulin-independent diabetes mellitus due to interactions between insulin-dependent diabetes mellitus and genetic causes, insulin resistance, and beta cell dysfunction, which are mainly caused by autoimmune processes. Because of the condition, absolutely dependent on insulin to prevent ketosis and death However, insulin-independent diabetes mellitus, which is called childhood diabetes, occurs in childhood, but the insulin secretion of the pancreas is relatively maintained, but insulin secreted by various reasons, such as obesity, lack of exercise, stress, and aging, can prevent adult diabetes. It is also known as the most common form of diabetes and accounts for over 90% of people with diabetes.

There are about 250 million people with diabetes worldwide, and the best goal of diabetic treatment is to bring the blood sugar of diabetics to normal blood sugar as much as possible, but in practice the normalization of blood sugar is a difficult task, and not only fasting blood sugar, The regulation of blood sugar (postprandial hyperglycemia) is increasingly emphasized (Jenkins, DJA, Wolever, TMS: Starchy foods and glycemic index.Diabetes Care ., 11 , 149-159; Menendez, CM, Stoecker, BJ: The role of diet in . improving glycemic control in Nutrition and diabetes :. Javanovic, L. and Peterson, CM (. ed), Alan R. Liss Inc, New York, p15-36 (1995)) for the regulation of blood sugar diabetes are currently sikhugwa diet The most effective drugs for postprandial hyperglycemia include insulin lispro, insulin analogues, and alpha-glucosidase inhibitors. There, we there is a majority of the carbohydrate in the dietary intake in the form of starch, the starch is absorbed after being digested by alpha amylase and alpha glucosidase. Therefore, the substance having alpha amylase or alpha glucosidase inhibitory activity can be expected post-prandial glycemic control effect. Acarbose, miglitol, and boglibose are used as drugs as alpha-glucosidase inhibitors that inhibit post-prandial hyperglycemia by inhibiting carbohydrate digestive enzyme activity in diabetics. Side effects occur frequently according to users, and therefore, it is necessary to develop safe blood glucose lowering agents with less side effects to improve the quality of life of diabetics and their families by appropriately treating diabetes, which is increasing at home and abroad.

Mugwort is an edible plant native to Korea, and is a food resource rich in protein, fat, fiber, saccharide calcium, phosphorus, vitamins A, B, and C. It has been widely consumed as foods such as herbs, mugwort, porridge, and mugwort rice cakes since ancient times. Among them, Injin is a perennial herbaceous deciduous shrub belonging to the family Asteraceae, Artemisia sacrum subsp. Kitamura, Artemisa sacrorum subsp.manshuria var.vestita Kitamura, etc. In particular, young leaves are called jinjin or heatjigi. Because of their unique fragrance and efficacy, they are traditionally used as edible, medicinal and medicinal herbs in the private sector. Has come. Injin is a young sprout of perennial perennial plant of Asteraceae, and the temper is tasteless, clear heat, moist, and yellowing. It is widely used in acute hepatitis, chronic hepatitis, cirrhosis, liver cancer, and is also used in diseases such as soap, gallbladder stones. Pharmacological action is to promote bile secretion, liver function protection effect, in particular the regeneration of liver cells. Lipid breakdown, coronary artery dilation and blood pressure lowering appear, antipyretic, diuretic, and antibacterial. Ji Ji-joon: Korean Pharmacopoeia and Korean Pharmacopoeia Standards Commentary, Seoul, Korea Medical Index, P638, 1998.) Injin mugwort has the effect of lowering heat and eliminating humidity. Its pharmacological action is known as its pharmacological action, as it inhibits liver damage (Ichiho et al., 2, Korean Society for Food Science and Nutrition, 22nd Autumn Conference, 1998), anticancer effect (Sung-kwon et al., Hakkuk Food Science Association) Conference Abstract, 1998), Antioxidant Activity, Blood Lipid Reduction and Liver Function Improvement Effect (Han Eun-kyung et al., 4, Hakkuk Food and Nutrition Science Spring Conference, 1997), Antimutagenic effect (Cho, Dong-seong et al., Journal of Biotechnology and Biotechnology, 2000) ), Antimicrobial effects (Jeong Mae-hee and Cho Yeon-hee, Injin mugwort, Sacheol mugwort, essential oil components and antibacterial effect of lion foot mugwort) are known. However, there have been no reports on the effects of Injin mugwort on postprandial blood sugar. In addition, processed products using phosphorus have been developed products such as raw mugwort syrup, flakes, pill or extract tea, granulated tea.

Therefore, the present inventors prepared extracts for dozens of herbal medicines for the purpose of developing post-prandial hypoglycemic drugs and measured the inhibitory effect of alpha-glucosidase, and found that the extracts of Injin mugwort had an excellent effect on postprandial blood sugar The present invention was completed.

As described above, the present inventors have tested the pharmacological effects on the extract obtained by extracting with ethanol from jinsui jinja to provide a substance having excellent postprandial glycemic inhibitory effect without any side effects in the living body, the post-prandial blood sugar in normal rats and diabetic rats It was confirmed that the lowering effect was excellent. In other words, the extract of Injin mugwort by the method of the present invention was found to have an effect of significantly reducing blood glucose concentration by inhibiting the activity of the alpha glucosidase enzyme.

Accordingly, it is an object of the present invention to provide a jinjin mugwort extract and a method for producing the same as a pharmacological composition for lowering blood sugar or a health functional food for lowering blood sugar. It is another object of the present invention to provide a composition containing the jinjin mugwort extract as an active ingredient.

EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.

The present invention is effective in inhibiting the digestion and absorption of carbohydrates after meals by controlling the metabolism of carbohydrates in humans and animals, a pharmaceutical composition comprising a jinjin wormwood and its preparation method and a pharmaceutically acceptable carrier using the extract as an active ingredient And diabetes treatment / prevention functional foods.

It is possible to use a known method as a production method of the extract of the present invention. For example, the extract of phosphorus mugwort may be added to solvents such as ethanol, ethanol, hexane, ethyl acetate, butanol, and water after being dried or dried, and then immersion extraction or warm extraction may be performed at room temperature. In the embodiment of the present invention, the extract obtained from the jinjin jinjin in butanol solvent was particularly excellent in hypoglycemic effect.

Injin mugwort extract of the present invention did not show any acute toxicity even when administered orally in large amounts in rats. In addition, since it is designated as a usable food ingredient of the Korea Food & Drug Administration, Injin mugwort extract has no problems such as side effects or toxicity, and can prevent or treat diabetes induction due to stress, excessive intake of fat, excessive intake of sugar and lack of exercise. It could be used as a therapeutic agent or as an active ingredient of functional foods.

The extract of Injin mugwort according to the present invention is considered to be a drug that can be safely administered without substantially any side effects or toxicity, and is formulated into a conventional pharmaceutical preparation according to a conventional method in the pharmaceutical field when the pharmaceutical composition is clinically used. It is available. In addition, the present invention jinjin wormwood extract may be used in the manufacture of functional foods that can lower the blood sugar in solid, liquid or jelly form by mixing in foods such as yogurt, beverages, ice cream, condiments, instant foods or livestock feed, in particular Since yogurt has a tendency to eat after meals, it can be easily used as a functional food for diabetic patients and diabetic prevention, which can prevent a sudden rise in blood sugar levels after eating in normal people or diabetics. The composition or functional food of the present invention may exert synergism by additionally including a known substance or a new substance used to prevent and / or treat diabetes by inhibiting glucose absorption of the human body in addition to the extract of Insulin mugwort if necessary.

Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to Examples. However, the following examples are only for the purpose of illustrating the present invention is not limited to the scope of the present invention.

Example 1: Preparation of Extract

Injin mugwort was purchased from Kyungdong Yangnyeong Market as a dry herb and used as extracting material. Phosphorus mugwort dried medicinal herbs and ethanol (ethanol) were mixed at a ratio of 1: 3 (W / V), respectively, and extracted by chilling for 7 days, filtered using a filter paper filter, and the filtrate was concentrated at 40 ° C. using a vacuum concentrator. The concentrate was re-dissolved in 70% methanol, and then extracted sequentially using four solvents (hexane, ethyl acetate, butanol, and water) according to the polarity, and concentrated at 40 ° C. using a vacuum condenser, and the water layer was freeze-dried. Lyophilization using was used as an extract sample.

Example 2: Screening of alpha-glucosidase inhibitory activity

The small intestinal epithelial layer of male Sprague-Dawley rats fasted for 16 hours was taken, suspended in 0.1 M phosphate buffer (pH 7.0), pulverized three times for 15 seconds by ultrasound, at 4 ° C and 10,000 rpm. The supernatant was used as a crude enzyme solution by centrifugation for 30 minutes. Glucose reagents to be used for the determination of alpha-glucosidase enzyme activity are 125 μg / ml O -phenylenediamine, 5 units / ml peroxidase and glucose oxidase (glucose oxidase) was used to make 0.96 units / ml. Coenzyme solution, 0.1 M phosphate buffer solution (pH 7.0), 2 mM maltose and sample solution were added at a final concentration of 20 ㎍ / ml at 37 ° C for 40 minutes. After incubation, the enzyme was inactivated in a constant temperature water bath (80 ° C., 3 minutes). Transfer 100 μl of the reaction solution to a new 96 well plate, add the same amount of glucose reagent, and then at 37 ° C. for 30 minutes. After incubation and color development, absorbance was measured at 490 nm by addition of 3 N hydrochloric acid (HCl). The inhibitory activity was determined by calculating the relative percent inhibition and 50% inhibition (IC50) compared with the positive control. It was.

Alphaglucosidase Inhibitory Activity of Extracts from Mugwort (microg / ml) extract Injin mugwort (IC ₅₀ ) Bokbunja (IC ₅₀ ) Peony (IC ₅₀ ) Ethanol extract 20.7 30 69 Hexane extract > 50 > 50 > 50 Ethyl acetate extract 18.6 39 16 Butanol extract 11.5 36 75 Water extract > 50 > 50 > 50

It was much better than Bokbunja (Korean Patent Publication No. 2003-0091178) or Peony (Korean Patent Publication No. 2003-0017092), which has been reported to be effective in the past. Particularly, butanol extract of Injin mugwort was the most effective. In addition, the ethanol extract was better than other medicinal extracts.

Example 3: Postprandial Blood Glucose Lowering Effect in Diabetic Rats

Twelve male Sprague-Dawley rats (between 80 and 120 mg / dl of blood sugar) weighed 250-300 g in a single injection of streptozotocin dissolved in 0.1 M citric acid buffer solution into the femoral muscles (75 mg / kg body weight) to cause diabetes. One week later, the animals were fasted for 12 hours and blood was collected from the tail, and blood glucose measured by a blood glucose meter (SUPER GLUCOCARDTM II, KDK) was found to be more than 200 mg / ml, indicating that diabetes was induced. The test animals were similarly divided into two groups (average 266.2 + 4.1 g, Injin mugwort group 269.5 + 4.3 g). In the diabetes-induced control group, maltose was orally administered with 2 g / kg of maltose, and blood glucose was measured at the tail vein at 0, 30, 60, 120, and 180 minutes postprandial. It was broken down into glucose by alpha-glucosidase in the small intestine and absorbed into the blood. After 60 minutes, the blood glucose level increased to 121 mg / dl, but it was administered with maltose. The 400 mg / kg oral administration of Injin mugwort extract with 400 mg / kg inhibited alpha-glucosidase activity, resulting in a significantly lower hypoglycemic effect. Therefore, ingestion of Injin mugwort extracts was significantly associated with postprandial blood glucose in diabetic rats. A synergistic inhibitory effect was shown.

Example 4 Postprandial Blood Glucose Lowering Effect in Normal Rats

Twenty male Sprague-Dawley rats weighing 120-140 g were weighed and divided into two groups with similar mean weights (control: 128.0 + 2.4 g, Injin mugwort group 130.0 + 2.8 g, mean + standard error). The solution administered to the animals was tested by oral administration in the same amount as the diabetic rats. At 0, 30, 60, 120 and 180 minutes after the meal, blood was collected from the tail vein and blood glucose was measured using a blood glucose meter. In the control group, maltose was orally administered with 2 g / kg, and then maltose was broken down into glucose by alpha-glucosidase in the small intestine of the rat and absorbed into the blood. The elevated blood sugar level was 56 mg / dl, but the wormwood butanol extract group administered with maltose inhibited alpha-glucosidase activity and blood glucose levels were 35 mg / dl after 30 minutes. Therefore, the intake of Injin mugwort extract showed a significant effect on suppressing postprandial blood sugar in normal rats.

Example 5 post-prandial blood sugar lowering effect of Injin mugwort yogurt

Sixteen male Sprague-Dawley rats, weighing 120-140 g, were divided into two groups with similar body weights (control group 130.0 + 3.3 g, Injin mugwort group 131.2 + 2.8 g, mean + standard error). Fermented into milk with yogurt using NUC lactic acid bacterium fermenter (ENU Electronics Co., Ltd.), mixed with 20% phosphorus mugwort extract and maltose 2 g / kg

Made yogurt Animals fasted for 12 hours were orally administered with an amount equivalent to 7.0 ml / kg body weight. Blood sugars were measured at the tail vein at 0, 30, 60, 120 and 180 minutes after the meal and measured by Glucotrend. In the case of Injin mugwort yogurt group, the glycemic value was 42 + 3.8 mg / dL at 30 minutes after the meal, which was significantly lower than the control group (61 + 5.1 mg / dl) (P <0.01). Ingestion of yoghurt yogurt significantly lowered postprandial blood glucose increase in normal rats.

As described above, the extract of the present invention Injin mugwort has a very excellent effect of inhibiting alpha-glucosidase enzymatic activity, inhibiting the digestion of carbohydrates after meals, suppressing the rapid rise of blood sugar, and preventing and / or treating diabetes. , Can be easily and safely control the postprandial hyperglycemia by adding to yogurt to be taken a lot after eating, Injin mugwort can be used as the main raw material of the food and drug safety notice notice of the food and beverages The present invention is a very useful invention in the pharmaceutical food industry.

Claims (3)

delete Food for inhibiting postprandial hyperglycemia, which comprises crude extracts extracted from jinseng wormwood with water, methanol or ethanol, or extracts extracted with ethyl acetate and butanol from the crude extracts A pharmaceutical composition for suppressing postprandial hyperglycemia, wherein the crude extract extracted from Ingeria mugwort with water, methanol or ethanol, or the extract extracted with ethyl acetate and butanol from the crude extract as an active ingredient

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