KR20210042718A - Composition for inhibiting glucose absorption containing Amomum villosum Loureiro as an active ingredient - Google Patents

Abstract

The present invention relates to a food composition for inhibiting glucose absorption containing an Amomum villosum Loureiro extract as an active ingredient, and a food composition for ameliorating gastrointestinal side effects containing an Amomum villosum Loureiro extract as an active ingredient. Accordingly, the gastrointestinal side effects such as abdominal distention, intestinal gas (flatulence, meteorism), and diarrhea which can occur in diabetes can be reduced, while ameliorating type 2 diabetes which is non-insulin-dependent diabetes.

Description

Composition for inhibiting glucose absorption containing Amomum villosum Loureiro as an active ingredient, containing Yangchunsa extract as an active ingredient}

The present invention relates to a composition for inhibiting glucose absorption containing Yangchunsa extract as an active ingredient and a food composition for improving gastrointestinal side effects containing Yangchunsa extract as an active ingredient. It relates to a composition with improved side effects.

Diabetes is a type of metabolic disease in which the secretion of insulin is insufficient or normal function is not performed. It is a disease that excretes glucose from urine and causes various complications due to high blood sugar.

Symptoms of diabetes are polyuria (increased urine volume, frequent urination), Daum syndrome (drinking a lot of water), and weight loss. If chronic hyperglycemia is maintained, development is reduced and infection is easily caused. .

If blood sugar is too high or too low, it can lead to loss of consciousness or coma due to ketoacidemia, hyperosmolar hyperglycemia syndrome, and hypoglycemia, and in severe cases, it can lead to an emergency situation that can lead to death.

Diabetes is a chronic disease that is difficult to cure, and complications can follow, and the representative ones are retinopathy, nephropathy, and neuropathy. In addition, the risk of cardiovascular diseases such as coronary artery disease, peripheral artery disease, and cerebrovascular disease is increased.

Diabetes mellitus causes complications in important parts of the body and increases mortality.

Therefore, to prevent complications due to diabetes, blood sugar control is being emphasized, and blood sugar is controlled through diet, exercise therapy, and drug therapy.

Diabetes is classified into type 1 and type 2, and type 1 diabetes is caused by destruction of pancreatic beta cells by autoimmune reactions, and type 2 diabetes is responsible for insulin secretion disorder due to insulin resistance and pancreatic beta cell malfunction. It is caused by

Here, the pancreatic β-cell is one of the cells of Langerhans Island, and it secretes insulin and promotes the inflow of glucose present in the blood vessels into the cells.

When the function of the pancreatic beta cells decreases, the secretion of insulin decreases, and blood sugar increases due to improper processing of glucose in the blood, leading to diabetes.

On the other hand, insulin resistance refers to the inability of cells to effectively burn glucose due to the lack of insulin function.Insulin receptors decrease due to abdominal obesity, lack of exercise, and excessive caloric intake, or the production of antibodies to insulin, or to insulin receptors. It is presumed to be generated by the generation of antibodies against the disease.

Diabetes treatment includes oral hypoglycemic drugs (insulin secretion stimulator, biguanide, alpha glucosidase inhibition, thiazolidinedione, etc.) administration or insulin injection along with diet and exercise therapy.

Insulin secretion stimulators can be further divided into sulfone urea drugs and non-sulfone urea drugs.

Although sulfone urea is the most widely used drug in the treatment of type 2 diabetes, hypoglycemia may occur as a side effect, so it should be selected according to the characteristics of the patient in consideration of the half-life, daily dose, and number of administrations per day between each sulfone urea agent.

The biguanide family increases insulin sensitivity, inhibits gluconeogenesis in the liver, lowers the rate of sugar production, and increases the absorption and utilization of sugar in the muscles.

However, biguanide drugs may appear as side effects related to the gastrointestinal tract, such as diarrhea, and lactic acidemia (a state in which the concentration of lactic acid (lactic acid) in the body has risen sharply) may appear, so kidney failure, liver disease, alcoholism, heart disease. In case of dysfunction or hypoxia, use should be avoided.

Alpha-glucosidase inhibitors reduce postprandial hyperglycemia by delaying the absorption of carbohydrates in the small intestine. For reference, alpha glucosidase is a type of glucosidase that breaks down carbohydrates.

Thiazolidinedione improves insulin sensitivity in the body and is involved in lipid metabolism and sugar metabolism in muscle or hepatocytes.

Taking thiazolidinedione may cause increased plasma volume and water retention, and may lead to edema or anemia, so it is contraindicated for use in heart failure or active liver disease.

In this way, there are side effects when using drugs, and it is difficult to improve and treat diabetes that requires long-term management.

In particular, acarbose is a type of alpha glucosidase inhibitor and is known to have side effects. The most common side effects are gastrointestinal symptoms such as abdominal pain, diarrhea, and gas frequently, which appear in proportion to the dose. In addition, there are cases where the level of liver function rises, so caution is required. Specifically, it is known that side effects such as abdominal distention, flatulence, meteorism, and diarrhea appear due to the effect of inhibiting pancreatic α-amylase (non-patent literature One). Inhibition of pancreatic alpha-amylase by acarbose increases the amount of undegraded starch in the colon, and the intestinal bacteria ferment the starch, releasing gas and low molecular weight substances (low-molecular weight). molecular-weight substances). It is reported that such abnormal fermentation is related to the side effects mentioned above (Non-Patent Document 2).

Accordingly, the applicant of the present invention studied natural substances that can reduce the side effects that may occur in diabetic patients, and as a result, using Amomum villosum Loureiro, side effects that may occur in insulin-independent (type 2) diabetic patients, specifically Found a composition capable of improving gastrointestinal side effects symptoms and completed the present invention.

Chiasson, J. L., Josse, R. G., Gomis, R., Hanefeld, M., Karasik, A., Laakso, M., & Group, S.-N. T. R. (2002). Acarbose for prevention of type 2 diabetes mellitus: the STOP-NIDDM randomised trial. Lancet, 359(9323), 2072-2077. doi: 10.1016/S0140-6736(02)08905-5. Dehghan-Kooshkghazi, M., & Mathers, J. C. (2004). Starch digestion, large-bowel fermentation and intestinal mucosal cell proliferation in rats treated with the alpha-glucosidase inhibitor acarbose. Br J Nutr, 91(3), 357-365. doi: 10.1079/BJN20031063.

The problem to be solved in the present invention is to provide a food composition and a pharmaceutical composition containing as an active ingredient a substance capable of reducing gastrointestinal side effects that may occur in the process of treating a type 2 diabetes patient.

In addition, the problem to be solved in the present invention is to provide a food composition and a pharmaceutical composition containing a natural substance, not a chemical synthetic substance, as an active ingredient.

The present invention for solving the above problems is a technical feature to provide a food composition for inhibiting glucose absorption containing Yangchunsa extract as an active ingredient.

In addition, the present invention for solving the above problems is a technical feature to provide a food composition for improving gastrointestinal side effects containing Yangchunsa extract as an active ingredient.

In addition, the present invention for solving the above problems is a food composition for improving gastrointestinal side effects containing Yangchunsa extract as an active ingredient, and the gastrointestinal side effects include abdominal distention, intestinal gas (flatulence, meteorism) and diarrhea. It is a technical feature to provide a composition of any one or more of (diarrhea).

In addition, the present invention for solving the above problems is a food composition for improving gastrointestinal side effects containing Yangchunsa extract as an active ingredient, the Yangchunsa extract at a concentration of 250 to 500 mg/kg based on rats. It is a technical feature to provide a composition for oral administration.

The food composition for inhibiting glucose absorption containing the Yangchunsa extract as an active ingredient and the food composition for improving gastrointestinal side effects containing the Yangchunsa extract as an active ingredient according to the present invention can occur in diabetes while improving type 2 diabetes, a non-insulin dependent diabetes mellitus. It can improve gastrointestinal side effects such as abdominal distention, flatulence, and diarrhea.

1 is a graph showing the result of alpha-glucosidase inhibitory activity
2 is a graph showing the result of alpha-amylase inhibitory activity
3 is a graph showing the results of sucrase inhibitory activity
4 is a graph showing the results of maltase inhibitory activity
5 is a graph showing the results of glucoamylase inhibitory activity
6 is a graph showing the results of a sucrose loading test
7 is a graph showing the amylose loading test result

Terms and words used in the present specification and claims conform to the technical idea of the present invention based on the principle that "the inventor can appropriately define the concept of terms in order to describe his or her invention in the best way." It should be construed as a meaning and concept, but not limited to a conventional or dictionary meaning.

Therefore, the embodiments described in the present specification and the configurations shown in the drawings are only the most preferred embodiments of the present invention, and do not represent all the technical spirit of the present invention, and thus various equivalents that can replace them at the time of application It should be understood that there may be water and variations.

COD (砂仁) means the overgrown antler four (綠殼砂Amomum villosum Loureiro var . Xanthioides TLWu et Senjen) or Amomum villosum chunks of ripe fruit or seeds (陽春砂. Amomum villosum Loureiro) over the ginger (Zingiberaceae) Thus, in the present invention, the fruit of Yangchunsa was used as a material.

Yangchunsa is mainly produced in Fujian, Guangdong, Guangxi, and Yunnan, and ankaksa is mainly produced in Indochina Peninsula areas such as Cambodia, India, Laos, Myanmar, Thailand, Vietnam, and some areas of Guangxi and Yunnan in China. China, v24:347-356.2000). Due to the different production areas, in ancient times, Nokgaksa was referred to as “Hokgaksa-in”.

Example. Preparation of Yangchunsa extract

Yangchunsa ( Amomum villosum ) was checked for authenticity in the herbal science classroom at Wonkwang University's Oriental Medicine College, and looking at the extraction process, 100g of Yangchunsa was heated with 900 ml of tertiary distilled water for 3 hours to extract under reflux, filtered, and the filtrate was concentrated under reduced pressure Freeze-dried to obtain 9.25 g of Yangchunsa extract.

Experimental Example 1. Rat intestinal α-glucosidase Inhibition assay ( In vitro )

1-1. Experimental process

After adding 100 mg of rat-derived intestinal acetone powder to 3 ml of 0.9% sodium chloride solution (NaCl solution), sonication in an iced water bath 12 times for 30 seconds, then centrifugation at 10,000 × g, 4℃ for 30 minutes Separated. Only the supernatant was used immediately for assay or was used while being stored at -20°C. 100 µl of a rat-derived alpha-glucosidase solution was poured over 50 µl of the sample solution and left to stand at 37°C for 10 minutes. 50 µl of 5 mM pNPG solution was added, reacted at 37° C. for 15 minutes, and absorbance was measured at 405 nm using an ELISA reader to analyze the rat-derived alpha-glucosidase inhibitory activity.

1-2. Experiment result

As shown in Fig. 1, the inhibitory activity of Yangchunsa extract against alpha-glucosidase derived from rats was 31.99±6.79%, 48.85±4.75%, and 62.58± at concentrations of 1, 3, and 5mg/ml, respectively. It was found to be 6.69%, showing a significant concentration-dependent inhibitory effect. In addition, as shown in FIG. 1, acarbose, known as an existing alpha-glucosidase inhibitor, also exhibited a concentration-dependent inhibitory effect.

For reference, alpha-glucosidase exists on the brush border of the small intestine, and plays a role in absorbing D-glucose by decomposing oligosaccharides and disaccharides. Alpha-glucosidase inhibitors reduce postprandial blood sugar (Krentz & Bailey, 2005).

_{The IC 50} for the inhibition of alpha-glucosidase of the extract of Yangchunsa according to the present invention was 3.287mg/ml as shown in Table 1 below.

Experimental Example 2. Porcine pancreatic α-amylase inhibition assay ( In vitro )

In the case of acarbose, not only has an alpha-glucosidase inhibitory effect but also has an inhibitory effect on alpha-amylase, so abdominal distention and intestinal gas (flatulence, meteorism) , Diarrhea, and the like are known to occur (Chiasson et al., 2002). Accordingly, the effect of Yangchunsa extract on alpha-amylase was confirmed.

2-1. Experimental process

200 µl of sample solution was added to 300 µl of Porcine-derived pancreatic α-amylase solution at a concentration of 1U dissolved in 0.02M sodium phosphate buffer (pH 6.9 with 0.006M sodium chloride) and incubated at 25°C for 10 minutes. To this solution, 500 µl of a 1% starch solution pre-incubated at 25°C for 10 minutes was added and reacted at 25°C for 10 minutes. The reaction was stopped by adding 1 ml of a 1% DNS solution dissolved in 30% Rochelle salt, followed by treatment in a boiling water bath for 5 minutes, then cooled to room temperature, and 10 ml distilled water was added. The reaction solution of the DNS solution and the sugar decomposed from the substrate by alpha-amylase was measured for absorbance at 540 nm using an ELISA reader.

2-2. Experiment result

As shown in Figure 2, in the group treated with the extract of Yangchunsa, Porcine-derived alpha-amylase did not show inhibitory activity against α-amylase. These results are similar to the inhibitory effect of acarbose on the activity of alpha-glucosidase, but suggest that Yangchunsa extract can be a candidate for the development of a new substance that can reduce its side effects.

Experimental Example 3. Rat intestinal glucose oxidase assay ( In vitro )

3-1. Experimental process

Rat-derived intestinal acetone powder (Sigma S9765) was used as the enzyme, and maltose, sucrose, and starch (Junsei) were used as substrates. After adding 100 mg of rat-derived intestinal acetone powder to 3 mL of 0.9% sodium chloride solution (NaCl solution, Junsei), sonication in an iced water bath 12 times for 30 seconds, then 10,000×g, 30 minutes at 4℃ Centrifuged. The separated supernatant was immediately used in the experiment.

As for the measurement method, 50 µl of a sample was added to 100 µl of rat alpha-glucosidase solution on a 96 clear plate, and then left in an incubator at 37° C. for 10 minutes. According to each experimental method, 50 µl of 100mM maltose, 200mM sucrose, or 1% starch solution was added, and then reacted at 37°C for 30 minutes. Between the reactions for 30 minutes, add 1 mL of a solution of a mixture of Glucose oxidase/peroxidase reagent (Sigma G3660) and O-Dianisidine reagnt (Sigma D2679) into a 2 mL tube (Tube, Eppendorf), and then stand in an incubator at 37°C for 5 minutes to raise the temperature to 37. After adjusting to °C, take 200ul of intestinal acetone powder, sample, and substrate solution mixed reagent from the rat that reacted for 30 minutes and add it to a solution of 1mL Glucose oxidase/peroxidase reagent and O-Dianisidine reagnt, and then in an incubator at 37°C for 10 minutes. The second reaction was carried out. After adding 1 mL of 12N sulfuric acid to each 2 mL tube (tube, Eppendorf) to stop the reaction, add 200 ul each to a 96 clear plate, and measure the absorbance at 540 nm using an ELISA reader to measure rat intestinal glucose oxidase (maltose, sucrose, glucoamylase) inhibitory activity was analyzed.

3-2. Experiment result

The inhibitory effect of the extract of Yangchunsa on sucrase, maltase, and glucoamylase of alpha-glucosidases was investigated.

Sucrase is an enzyme that decomposes sucrose, a disaccharide, into glucose and fructose, which are monosaccharides, 1, 3, 5mg/ml Yangchunsa extract as shown in FIG. In the treated group, the inhibitory effects on sucrase activity were 39.01±5.96%, 53.68±2.21%, and 93.21±3.78%, respectively, showing an inhibitory effect similar to that of acarbose. This result is thought to be helpful in suppressing the increase in blood sugar due to the intake of sucrose, a major added sugar in food.

Maltase is an enzyme that breaks down maltose, a disaccharide, into two molecules of glucose. As shown in Figure 4, in the group treated with 1, 3, 5mg/ml Yangchunsa extract, the inhibitory activity against maltase was 32.87±1.86%, 34.62±6.06%, and 58.36±1.30%, respectively. These results suggest that Yangchunsa extract will help inhibit the activity of maltase, one of the disaccharide-degrading enzymes.

Glucoamylase (glucoamylase) is one of the alpha-glucosidases (α-glucosidases), as an enzyme having glucan 1,4-alpha-glucosidase activity (glucan 1,4-α-glucosidase activity), shown in Figure 5 As shown, in the group treated with 1, 3, 5mg/ml Yangchunsa extract, the inhibitory activities were 20.61±2.49%, 23.05±5.24%, and 56.07±4.26%, respectively. These results suggest that Yangchunsa extract is effective in inhibiting glucoamylase activity.

_{As shown in Table 1, IC 50} values for inhibition of sucrase, maltase, and glucoamylase activity were 2.117, 4.263, and 4.890 mg/ml, respectively.

Experimental Example 4. Sucrose and amylose Loading test ( In vivo )

As an extension of the experiment to confirm the inhibitory effect of Yangchunsa extract on α-glucosidase activity in vitro , a sucrose loading test was performed using SD rats in vivo.

4-1. Experimental process

After fasting the experimental animals for at least 20 hours before the experiment, the body weight was measured and divided into 10 animals by group (control group, positive control group, test group), and then sucrose calculated with a dose of 2.0 g/kg-body weight relative to the body weight. ), amylose and Yangchunsa extract (0.25g/kg-bw, 0.5g/kg-bw) are precisely weighed and dissolved in 1 mL of distilled water. In the case of the control group, 1 mL of distilled water was prepared, and in the case of the positive control group, acarbose was calculated as 5.0 mg/kg-bw (Glucobay, Bayer Korea) and weighed. To make a solution.

Prepared control (distilled water 1mL), test group (0.25g/kg-bw, 0.5g/kg-bw sample + 2.0 g/kg-body weight concentration of sucrose and amylose-containing distilled water 1 mL}, positive control {acarbose 5.0 mg/kg-bw + 2.0 g/kg-body weight of sucrose and 1 mL of distilled water containing amylose} Samples were administered orally at 1 ml/ml using a zone for oral administration. , After oral administration, blood is collected from the tail vein for 30 minutes, 60 minutes, and 120 minutes, and the change in blood glucose concentration of venous blood is measured with a blood glucose kit (Caresens II).

4-2. Experiment result

2.0g/kg sucrose was mixed with 5mg/kg acarbose (positive control) and Yangchunsa extract at a concentration of 250, 500mg/kg and administered orally, and blood glucose was measured after 0.5, 1, and 2 hours. As a result, as shown in Fig. 6, the blood sugar in the acarbose-administered group after 30 minutes was 95.67±12.88mg/dL, 250, 500mg/kg Yangchunsa extract-administered group was 155.17±16.36 and 150.33±13.17mg/dL, respectively It showed a significant reduction effect (p<0.05) compared to the control group (183.17±13.50mg/dL). In addition, after 60 minutes, blood glucose was also significantly decreased (p<0.05) compared to the control group. These results are thought to be the result of in vivo confirmation of the inhibitory effect on the activity of α-glucosidase of Yangchunsa extract verified in vitro.

To investigate the effect of Yangchunsa extract on pancreatic α-amylase activity under in vivo conditions, 2.0g/kg amylose was used as a positive control, 5mg/kg acarbose and 250, 500mg. As a result of measuring blood glucose 0.5, 1, and 2 hours after oral administration by mixing with /kg concentration of Yangchunsa extract, blood glucose 30 and 60 minutes after acarbose administration was 73.33±5.12, respectively, as shown in FIG. , 80.0±8.88mg/dL, significantly decreased compared to the control group (114.17±23.11, 117.83±17.15mg/dL, respectively), indirectly suggesting that α-amylase activity was inhibited.

However, in the group administered with Yangchunsa extract, there was no significant difference in blood glucose compared to the control group, and thus the inhibitory activity of alpha-amylase was low even under in vivo conditions, corresponding to the in vitro results (Fig. 1B).

Claims (6)

Food composition for inhibiting glucose absorption containing Yangchunsa extract as an active ingredient.
Food composition for improving gastrointestinal side effects of diabetic patients containing Yangchunsa extract as an active ingredient.
The method according to claim 2,
The gastrointestinal side effects are abdominal distention, intestinal gas (flatulence, meteorism), and diarrhea (diarrhea), characterized in that any one or more of the gastrointestinal side effects improvement food composition for diabetic patients.
The method according to claim 2,
The Yangchunsa extract is a food composition for improving gastrointestinal side effects of diabetic patients, characterized in that oral administration at a concentration of 250 to 500 mg/kg based on rats.
A pharmaceutical composition for preventing or treating gastrointestinal side effects of diabetic patients containing Yangchunsa extract as an active ingredient.
The method of claim 5,
The gastrointestinal side effects are abdominal distention, intestinal gas (flatulence, meteorism) and diarrhea (diarrhea), characterized in that any one or more of the gastrointestinal side effects prevention or treatment pharmaceutical composition for diabetic patients.

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