KR20090120738A - Composition for treating diabetes mellitus and obesity containing the powder of poncirus trifoliata's peel - Google Patents

Abstract

PURPOSE: A composition for treating and preventing diabetes containing Poncirus trifoliatas skin is provided to suppress α-glucosidase and reduce body weight and blood sugar. CONSTITUTION: A composition for preventing and treating diabetes contains Poncirus trifoliatas skin powder. The composition suppresses the activation of α-glucosidase. A method for producing Poncirus trifoliatas skin powder comprises: a step of freeze-drying Poncirus trifoliatas skin; a step of pulverizing the Poncirus trifoliatas skin; a step of adding spirit then extracting 24 hours; a step of filtering the extract then adding spirit; and a step of decompress-filtering the extract.

Description

Composition for treating diabetes mellitus and obesity containing the powder of Poncirus trifoliata's peel}

The present invention relates to a composition that can be usefully used for the prevention and treatment of diabetes mellitus containing tanza bark powder as an active ingredient, and more particularly, diabetes mellitus using a significant hypoglycemic effect and weight loss effect of the tanza bark powder It relates to a composition for the prevention and treatment of obesity and improvement of obesity.

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. (Abrams, JJ, Ginsberg, H, et al., Diabetes , 31 , pp 903-910, 1982).

Diabetes mellitus has two types of characteristics. Type 1 diabetes mellitus is caused by a lack of secretion of insulin, a glucose-regulating hormone in the blood, mainly younger people in their 10s to 20s. It is also called juvenile diabetes because it occurs in. Type 2 diabetes mellitus usually occurs 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 that it is caused by genetic factors and environmental factors involved. As a etiology of type 2 diabetes, both pancreatic beta-cells with insulin secretion and target cells with insulin action defects (insulin resistance) are observed.

The most important goal in the treatment of diabetes is to control blood glucose levels as close to normal as possible, and postprandial blood sugar control, together with fasting blood glucose, is important for the prevention and treatment of diabetic symptoms and complications (Jenkins, DJA, Wolever et al., Starchy foods and glycemic index, Diabetes Care , 11 , pp 149-159, 1988; 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., pp 15-36, 1995), Methods of Treatment Drug therapy, diet and exercise therapy.

Oral hypoglycemic agents currently used in type 1 and type 2 diabetics include α-glucosidase inhibitors, sulfonylureas, and biguanides. Glucosidase 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 glucose and blood insulin. α-glucosidase inhibitors promote the secretion of glucagon-like peptide-1 in the small intestine that promotes insulin secretion and suppresses glucagon secretion without causing hyperinsulinemia or hypoglycemia Have advantages (Mooradian, AD, Thurman, JE, Drugs , 57 , pp 19-29, 1999; Baron, AD, Diabetes Research and Clinical Practice , 40 , pp S54-S55, 1998).

The α-glucosidase inhibitors currently used in the clinic include acarbose (acarbose), bogillibose and miglitol (miglitol), and the current marketed acabose as an α-glucosidase inhibitor to diabetic animals Fasting blood glucose and glycated hemoglobin levels decreased (Wright BE, Vasselli JR, Katovich MJ. Phys Behavior, 63, pp 867-874, 1998), and fasting blood glucose levels decreased when ingested acarbose in patients with type 2 diabetes. (Hanefeld M, Fischer S, Julius U, et al. Diabetologia, 39, pp 1577-1583, 1996). In addition, Lebovitz (Lebovitz, HE Diabetes, Rev. 6, pp 132-145, 1998) reported that ingestion of acarbose inhibits post-prandial blood sugar levels and decreases glucose toxicity, thereby reducing fasting blood glucose. Cheonggukjang, a fermented soybean meal, was found to significantly reduce fasting blood glucose levels in animals induced with type 2 diabetes by administering streptozotocin at 5 days of age (Kang MJ, Kim JI, Kwon TW. Food Sci Biotechnology, 12 , pp 544-547, 2003). In a study by Fujita et al. (Fujita H, Yamagami T. Life Science, 70, pp 219-227, 2001), after long-term intake of Touchi water extract, a traditional Chinese fermented soybean meal, to KKAY mice, Fasting blood glucose was also reported to be significantly reduced. In addition, the intake of acarbose in obese and diabetic animals was reported to significantly reduce body weight (Yamamoto M, Jia DM, Fukumitsu KI, Imoto I, Kihara Y, Hirohata Y, Otsuki M. Metabolism, 148, pp 347-354 , 1999).

However, long-term use of α-glycosidase inhibitors may have side effects, such as abdominal bloating and vomiting diarrhea in some patients, which may limit their use (Hanefeld, M., Journal of Diabetes and its Complications , 12 , pp 228-237, 1998). Therefore, researches on blood glucose control substances that have no side effects from natural products are actively conducted.

On the other hand, the average lifespan of erythrocytes is 120 days. In diabetic patients, non-enzymatic and irreversible glycosylation of hemoglobin in erythrocytes continuously occurs, leading to increased glycated hemoglobin levels (Tanaka Y, Uchino H, Shimizu T. European J.). Phar, 376, pp 17-22, 1999: Ahn HS, Lim EY, Kim HR.J Korean Soc Food Sci Nutr, 26 (5), pp 914-919, 1997). Therefore, glycated hemoglobin concentration has been reported to be more meaningful to investigate the long-term glycemic control effect than fasting blood glucose. KKAY mice, a type 2 diabetic animal model, exhibited accumulation of saccharified products as the age of the age increased, and accelerated as the diabetic progressed, thereby increasing the glycated hemoglobin concentration. However, these changes were suppressed when the experimental diet was ingested for a long time, resulting in glycated hemoglobin. The concentration is reduced (Ahn HS, Lim EY, Kim HR. J Korean Soc Food Sci Nutr, 26 (5), pp 914-919, 1997).

Many studies have reported that decreased glycated hemoglobin levels, which indicate long-term glycemic control, can prevent diabetes complications. A 2% reduction in glycated hemoglobin levels in type 2 diabetic patients resulted in a 68% and 70% reduction in retinopathy and neuropathy, respectively (Ohkubo Y, Kishikawa H, Araki E, et al. Diabetes Res Clin Pract, 28, pp. 103-117, 1995). A 1% reduction in glycated hemoglobin levels reduced microvascular complications by 25% after 10 years and a 16% reduction in myocardial infarction risk (United Kingdom Prospective Diabetes Study Group. Lancet 352, pp 837-853, 1998). The importance of glycemic control is shown to reduce vascular complications. In diabetic retinopathy of type 2 diabetic patients, when the blood glycated hemoglobin concentration increased from 9% to 11%, the risk of retinopathy doubled (Klein R, Klein BEK, Moss MD). JAMA 260, pp 2864-1871, 1988).

Poncirus trifoliata ) belongs to the phyto taxonomy of the Raceae family. Tenza is more cold-resistant than other citrus fruits, so it can grow to sub-Gyeonggi, has a distinctive scent and sour taste, and is mainly dried and used as a herbal medicine (Chung HS, Lee JB, Seong JH, Choi JU.Korean Journal of Food Preservation, 11 (3), pp 342-346, 2004). As for the efficacy of the tanza, the functions of anti-inflammatory, anti-intestinal bacteria and anti-cancer were studied (Kim HM, Kim HJ, Park ST. J. Ethnopharmacol , 66, pp 283-288, 1999: Yi JM, Kim MS, Koo). HN, Song BK, Yoo YH, Kim HM.Clinica Chimica Acta , 340, pp 179-185, 2004).

However, there are no studies on the glycemic control effects of tangerine shells.

The present inventors have completed the present invention by confirming that the tanza bark has an improvement effect on the metabolism of the metabolism, has the effect of preventing and treating diabetes and diabetic complications due to the disturbance of the metabolic metabolism, and also has a weight loss effect.

It is therefore an object of the present invention to provide a composition for the prevention and treatment of diabetes and the improvement of obesity.

In order to achieve the above object, the present invention provides a composition for preventing and treating diabetes containing tanza bark powder.

The composition inhibits the activity of α-glucosidase, reduces plasma glucose concentration, and decreases the concentration of blood glycated hemoglobin.

In addition, the present invention provides a composition for improving obesity containing tanza bark powder.

The tanza shell of the present invention not only shows excellent α-glucosidase inhibitory activity, but also slows the digestion rate of carbohydrates after meals, inhibits the rapid rise of blood glucose levels and decreases the concentration of glycated hemoglobin, and together with the effect of reducing weight. Therefore, the composition comprising the same may be usefully used as a pharmaceutical and nutraceutical for the prevention and treatment of diabetes and obesity.

The present invention provides a composition that can be usefully used for the prevention and treatment of diabetes and obesity by containing a tanza bark powder having a hypoglycemic effect.

The tanza shell contained in the composition of the present invention is preferably pulverized and pulverized in accordance with conventional methods known in the art to use the powder.

The content of the powder component depends 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 of time, which may be appropriately selected by those skilled in the art.

The composition of the present invention reduces the fasting blood sugar and glycated hemoglobin, provides the effect of weight loss, and thus can be used as a pharmaceutical composition or dietary supplement composition for the prevention and treatment of diabetes and obesity.

When the composition according to the present invention is used as a pharmaceutical composition, the preferred application amount of the composition depends 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.

In addition, the composition of the present invention may contain one or more components exhibiting the same or similar function together with the tangerine shell. In addition, the composition of the present invention may be prepared by additionally containing one or more pharmaceutically acceptable carriers for administration, saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol and ethanol One or more selected from the group consisting of can be used as a pharmaceutically acceptable carrier. It may also be formulated into aqueous solutions, pills, capsules, granules or tablets, if desired, using other conventional additives such as antioxidants, buffers, bacteriostatics, diluents, dispersants, surfactants, binders or lubricants.

In addition, when using the composition according to the present invention as a nutraceutical composition, the tanger bark may be used alone or in combination with other foods, food additives, may be appropriately used in accordance with conventional methods. The mixing amount of the tanger bark may be appropriately determined depending on the purpose of use, such as health, prevention of disease or therapeutic treatment.

Hereinafter, preferred test examples are presented to help understand the present invention. However, the following test examples are merely provided to more easily understand the present invention, and the content of the present invention is not limited to the test examples.

Reference Example 1 Preparation of Tangerine Peel Extract

Commercially purchased (purchased from: farmer's market in Eomgung-dong, Busan) tanza shells were lyophilized and then ground and used as samples for the experiment. The general components of tanza bark were analyzed by AOAC method, and moisture, crude protein, crude fat, crude ash and total fiber content were measured. Moisture was analyzed by atmospheric pressure drying, crude fat by Soxhlet extraction, crude protein by semi-micro Kjeldhal method, and crude ash by ingestion method (Food Code (2005) , Korea Food Research Institute). Total dietary fiber content was measured by AOAC method developed and modified by Prosky et al. (Prosky, L., et al., Journal-Association of Official Analytical Chemists , 71 , pp1017-1023, 1988). The content of common ingredients was 5.1g of water, 6.8g of fat, 6.3g of protein, 4.0g of ash, and 45.6g of dietary fiber per 100g.

After freeze-drying, 15 g of liquor was added to 100 g of the ground tanza husk crushed and extracted for 24 hours, filtered under reduced pressure with a filter paper (Watman, USA), followed by 5 liters of liquor, followed by extraction for 12 hours. After filtration under reduced pressure), the filtrates were collected and concentrated under reduced pressure at 37 ° C. using a vacuum rotary concentrator to obtain 33.4 g of alcohol extract, and stored at −20 ° C.

[Test Example 1] in vitro ( in vitr o) Determination of α-glucosidase inhibitory activity

Yeast α-glucosidase used in this experiment was purchased from Sigma, Louis, MO, USA, and the enzyme substrate was used as para-nitrophenyl-alpha-D-glucopyranoside. As a control group, acarbose (trade name: Glucoby, Bayer-Korea), which is used clinically, is used as a blood glucose increase inhibitor. After drying the tanza bark extract obtained in Reference Example 1, and dissolved in dimethyl sulfoxide at a concentration of 0.5 mg / ㎖ to prepare a solution, it was an enzyme reaction solution containing yeast (alpha)-glucosidase To the degree of inhibition of α-glucosidase. The measurement was repeated three times, and the results are shown as mean ± standard deviation in Table 1 below.

α-glucosidase inhibitory activity Concentration (mg / ml) Tanza bark extract Akabos 0.50 26.2 ± 1.4% 35.0 ± 0.9%

In Table 1, the inhibitory effect of tanza bark extract and acarbose at concentrations of 0.5 mg / ml was 26.2 ± 1.4% and 35.0 ± 0.9%, respectively, and the components of the tanza bark significantly decreased α-glucosidase. It can be confirmed that the inhibition.

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

In vitro in Test Example 1 ( In vitro) α- glucosidase vivo (In the taengja shells appeared to be excellent in inhibitory activity In vivo ) To measure the effects of diabetes mellitus and diabetic complications, db / db mice of type 2 diabetic animal model were divided into three groups of seven animals each, and the average blood glucose levels of the two groups (Examples 1 and 2) were similar. AIN-93G based diet shown in Table 2 was used for the comparative example, which was separated by the egg mass method, and 5% (Example 1) and 10% of the tanza shells lyophilized to the basic diet, for the experimental groups, Examples 1 and 2, shown in Table 2 below. (Example 2) The diet containing was ingested for 6 weeks. Diet and drinking water freely ( ad libitum ), the temperature and humidity of the breeding room were maintained at 20 ~ 25 ℃, 50 ~ 60%, respectively, the contrast was turned on and off every 12 hours. Animal weight and dietary intake were measured twice and three times a week, respectively.

Diet  Control diet Diet containing tangerine shell powder Example 1 (5%) Example 2 (10%) Cornstarch ¹⁾ 39.8 39.8 39.8 Gelatinized starch 13.2 13.2 13.2 Casein ^{2 )} 20.0 19.7 19.4 Soybean oil ^{3 )} 7.5 6.7 6.3 α-cellulose ^{4 )} 6.3 2.7 0.4 Sucrose ^{3 )} 10.0 8.4 6.8 Mineral mixtures ^{5 )} 3.5 3.5 3.5 Vitamin mixtures ^{6 )} One One One L-cystine ^{2 )} 0.3 0.3 0.3 Choline bitartrate ⁴⁾ 0.25 0.25 0.25 T-butylhydroquinone ^{7 )} 0.0014 0.0014 0.0014 Tangerine peel powder ^{8 )} 0 5 10  ) Daesang, Korea 2) ICN Biochemical, USA 3) Cheil Jedang, Korea 4) Sigma, USA 5) AIN-93G Mineral mix., ICN Pharmaceuticals Inc., U.S.A. 6) AIN-93G vitamin mix., ICN Pharmaceuticals Inc., U.S.A. 7) Fluka Co., U.S.A. 8) Freeze-dried and milled

Animals were sacrificed at 6 weeks of diet. The animals were fasted for 12 hours before sacrifice and suffocated with carbon dioxide gas and blood was collected from the heart with a syringe containing 10 mg of EDTA (ethylene diamine tetra acetic acid). Blood was centrifuged at 3,000 rpm for 15 minutes to collect plasma and stored at -70 ° C.

All results were expressed as mean ± standard deviation. Significance test between control group and tanza bark group was carried out using ANOVA and Tukey's test was used as a post test (α = 0.05).

Test Example 2 Measurement of Dietary Intake and Weight Gain

The weight and dietary intake of the mice after the diet for 6 weeks for the Comparative Example, Example 1 (5% tanza bark extract) and Example 2 (10% tanza bark extract) are shown in Table 3.

Comparative example Example 1 Example 2 Weight (g) 40.45 ± 1.83 ^a 35.72 ± 2.86 ^b 33.87 ± 3.94 ^b Dietary Intake (g / day) 5.22 ± 0.32 ^a 4.64 ± 0.40 ^b 4.21 ± 0.49 ^b  * Mean ± S.D * Significant difference when superscript is different (p <0.05)

As shown in Table 3, the weight of the Comparative Example, Example 1 and Example 2 was 40.45 ± 1.83 g, 35.72 ± 2.86, 33.87 ± 3.94 g, respectively, and the dietary intake of Comparative Example, Example 1 and Example 2 Is 5.22 ± 0.32 g / day, 4.64 ± 0.40 g / day, 4.21 ± 0.49 g / day, respectively, it can be seen that the weight and dietary intake of Example 1 and Example 2 showed a significant difference compared to the comparative example.

Therefore, it can be inferred from the above results that ingestion of tangerine shell may reduce body weight and improve obesity.

Test Example 3 Measurement of Plasma Glucose Concentration Improvement Effect of Tangerine Peel

Glucose using the glucose (glucose) content in the fasting plasma assay kit (assay kit, Sigma, USA) oxidase (glucose oxidase), enzyme method (Raabo E, Terkildsen TC, On the enzymatic determination of blood glucose. Scand J Clin Lab Invest 12: 402, 1960), and the results are shown in FIG.

As shown in FIG. 1, the blood glucose of Example 1 and Example 2 was 391.4 ± 34.9mg / dL, 358.1 ±, after ingesting tanza bark with excellent α-glucosidase inhibitory activity to type 2 diabetic animals for 6 weeks. 40.0 mg / dL significantly reduced fasting blood glucose compared to the comparative example (473.4 ± 41.6 mg / dL) (p <0.05).

Therefore, it can be seen from the results that the long-term intake of the tanza shell of the present invention can significantly reduce fasting blood sugar, and can be effective in improving and treating diabetes mellitus.

Test Example 4 Determination of Blood Glycosylated Hemoglobin Concentration of Tanza Husks

Glycated hemoglobin concentration was measured by column chromatography. 50 μl of hemolyzing reagent (Hemolyzing reagent, BioSystem, Spain) was added to 50 μl of blood, and 50 μl of the hemolytic sample obtained by standing at room temperature for 10 minutes was placed in a column filled with cation-exchange resin. The base wall was washed with 200 µl of washing reagent, 2 mL of the same reagent was drained through the column, and the fraction eluted with 4 mL of eluting reagent was recovered. Then, 12 mL of the eluent was added to 50 μl of the hemolytic sample to prepare total Hb. After measuring the absorbance of the total Hb with and distilled water in the spectrophotometer 415nm as a control, the glycated hemoglobin value was calculated by substituting Equation 1 below, and the results are shown in FIG. 2.

A = absorbance of HbA _1c

B = absorbance of total Hb

As shown in FIG. 2, the results of ingesting the tanza bark into the diabetic animals of type 2 for 6 weeks, the blood glycated hemoglobin concentrations of Examples 1 and 2 were 7.0 ± 0.7% and 6.6 ± 0.5%, respectively. 0.7%), and significantly decreased (p <0.05).

Therefore, it can be inferred from the above results that the tanza shell of the present invention has a glycemic control effect and may be useful for preventing diabetes complications.

1 is a graph showing the blood glucose lowering effect of Comparative Example, Example 1 and Example 2.

Figure 2 is a graph showing the glycated hemoglobin lowering effect of Comparative Example, Example 1 and Example 2.

Claims (5)

A composition for preventing and treating diabetes containing tanza bark powder. The composition for preventing and treating diabetes of claim 1, wherein the composition inhibits the activity of α-glucosidase. The composition for preventing and treating diabetes of claim 1, wherein the composition reduces plasma glucose concentration. The composition for preventing and treating diabetes of claim 1, wherein the composition reduces the concentration of blood glycated hemoglobin. Composition for improving obesity containing tanza bark powder.

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