KR101350826B1 - Composition for the prevention and treatment of diabetes mellitus comprising extracts or fractions of eremochloa ophiuroides as an active ingredient - Google Patents

Abstract

The present invention relates to pharmaceutical compositions for preventing or treating diabetes mellitus, comprising an extract of centipede grass (Eremochloa ophiuroides) as an active ingredient. The present inventors verified that an extract from centipede grass, a fraction thereof, and an active fraction isolated from the fraction effectively adjusted the blood glucose concentration. Therefore, the pharmaceutical compositions of the present invention can be useful as a composition for preventing or treating diabetes mellitus, a functional food for preventing or treating diabetes mellitus, a composition for lowering and adjusting the blood glucose level, and a health food for lowering and adjusting the blood glucose level.

Description

Composition for the prevention and treatment of diabetes mellitus comprising extracts or fractions of Eremochloa ophiuroides as an active ingredient}

The present invention relates to the use of Centipede grass ( Eremochloa ophiuroides ) relates to a composition for preventing and treating diabetes containing the extract or fractions thereof as an active ingredient.

Recently, Korea also shows a very desirable phenomenon in terms of physical development due to rapid economic growth, improved living standards, and westernization.In the case of adults, the disease pattern is caused by stress caused by excessive high calorie food intake, lack of exercise and industrial society. Increasingly, the disease is becoming more westernized, especially in adult diseases. Representative adult diseases include liver disease, high blood pressure, diabetes, obesity, hyperlipidemia and the like.

Diabetes, one of the typical adult diseases, is a non-communicable chronic disease. Insulin secreted by the pancreatic β cells is insufficient or does not function properly. Therefore, glucose is not used as an energy source in the body and remains high in the blood. It is a disease that is excreted. Diabetes can be broadly classified into insulin dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM) according to its pathogenesis and treatment of symptoms. In Korea, more than 95% of diabetics are NIDDM patients. Diabetes is not a big disease in itself but rather a complication caused by long-term disease, such as diabetic neuropathy, retinopathy, cataract, nephropathy, etc. This is a social problem because patients can not only live a normal life but can have fatal consequences.

Currently used diabetic drugs are roughly divided into oral hypoglycemic agents and insulin injections. In general, insulin injections for insulin-dependent diabetics without pregnancy in the body, gestational diabetes patients, and insulin-independent diabetes patients whose oral hypoglycemics are poorly controlled are given adequate insulin injections, despite the combination of diet and exercise therapy. Insulin-independent diabetic patients who do not have blood sugar control are generally allowed to take oral hypoglycemic agents, and oral hypoglycemic agents may be classified into sulfonylurea drugs and biguanide drugs.

Sulfonylurea-based drugs such as glipizide, gliclazide, gliquidone, glibenclamide, and chlorpropamide have been shown to promote insulin secretion in the pancreas. However, it cannot be used for insulin-dependent diabetics who have no insulin secretion in the pancreas and cannot be used for women of childbearing age due to concerns about insulin-independent diabetes patients with relatively reduced insulin secretion in the pancreas, birth defects, miscarriage, and stillbirth. There are disadvantages. In addition, since most sulfonylurea-based drugs are metabolized in the liver and excreted into the kidney, they should be used with caution in patients with hepatic and renal function disorders.

Although biguanide-based drugs such as metformin have no clear mechanism of action, they have not been shown to increase insulin secretion in the pancreas, and have a lower hypoglycemic effect than sulfonylurea-based drugs. low. However, due to the high incidence of side effects of the digestive system, nausea, vomiting, diarrhea and rash appear at the beginning of treatment, and cause lactic acidosis and cause life-threatening side effects. have.

Therefore, currently used sulfonylurea-based or biguanide-based drugs have the above disadvantages or side effects, and therefore, there is a need to develop a safer hypoglycemic agent with less side effects and more desirable to treat diabetic diseases that are increasing at home and abroad. There is.

As described above, since the serious effects of chemically synthesized antidiabetics and the drug effects due to the development of drug resistance have been declining, it is necessary to avoid chemosynthetic antidiabetics and find new substances with new antidiabetic activity from natural products. Many studies are in progress.

Sentipedgrass ophiuroides ; Centipede grass; Nanji turf) is a late-turf turf that is a perennial grass that crawls and forms a turf. Belongs to Poaceae, scientific name is Eremochloa ophiuroides (Munro) Hack. It is mainly distributed in China, East Asia, Indochina, Northeastern USA, Mesoamerica and the Caribbean. It is well adapted to various soil conditions, but it is particularly resistant to moist and acid soil and low fertility soil. As a morphological feature, the leaf has a length of 15 ~ 30 ㎜, a width of 2 ~ 4 ㎜, a flat shape with a mid vein, no hairs except a collar part, and a leaf tip round. The stem is a branching type stolons with a very firmly sheath (sheath). The flower part has a three to five inches long fossil flower (raceme), which has a purple color and a slightly flat, spikelet arranged in two lines. I have. It is used as a lawn or turf grass and is used as a forage for its leafiness and good taste.

Sentipedgrass (warm turf grass) has been researched for the purpose of preventing and treating obesity, preventing and treating cancer, preventing skin aging, skin whitening and the like. However, there have been no specific and systematic studies on which pharmacological effects among the various components of centipedegrass have been reported. In particular, there are no reports regarding the therapeutic efficacy of diabetic diseases.

Therefore, while the present inventors are studying natural plant extracts having excellent anti-diabetic activity, the extracts extracted from Sentedgrass, fractions thereof, and active fractions separated from the fractions can be effectively used as new diabetes therapeutic agents by effectively controlling blood glucose levels. The present invention has been completed by revealing that there is.

An object of the present invention is to provide a composition for the prevention and treatment of diabetes mellitus containing Centipede grass ( Eremochloa ophiuroides ) extract or a fraction thereof as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the prevention and treatment of diabetes containing Centipede grass (Centipede grass, Eremochloa ophiuroides ) extract as an active ingredient.

In order to achieve the above object, the present invention provides a functional food for preventing or improving diabetes containing centipede glass extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for controlling blood sugar drop containing centipede glass extract as an active ingredient.

In order to achieve the above object, the present invention provides a health food for controlling blood sugar drop containing centipede glass extract as an active ingredient.

Centipedgrass extract, fractions thereof, or active fractions separated from the fractions of the present invention can be usefully used as a novel diabetic prevention or treatment composition, or as a functional food by effectively controlling blood glucose concentrations as non-toxic natural products.

1 is a schematic diagram showing a process of preparing an extract of Sentipedgrass, fractions thereof, and active fractions separated from the fractions of the present invention.
FIG. 2 is a diagram showing the amount of fat accumulation in FIG.
Figure 3 is a diagram showing the degree of fat accumulation produced by the glucose metabolism process of fat cells by the centipede glass ethyl acetate- methanol fraction.
Figure 4 is a picture confirming the hypoglycemic activity in type 2 diabetic mice by the centifed glass ethyl acetate- methanol fraction.
Figure 5 is a picture confirming the results of the anti-diabetic experiment in type 2 diabetes mice by centifed glass ethyl acetate- methanol fraction.
6 is a diagram confirming the change in blood glucose by oral glucose experiment in type 2 diabetes mice.
FIG. 7 shows the results of glucose oral loading experiments using the doses of the centifed glass ethyl acetate-methanol fraction at 1 mg / kg and 10 mg / kg.
8 is a diagram showing the results of the intraperitoneal sugar load test 1 mg / kg and 10 mg / kg dose of the centifed glass ethyl acetate-methanol fraction.
Figure 9 is a diagram confirming the change in blood glucose by intraperitoneal administration of the centifed glass ethyl acetate- methanol fraction and then oral glucose loading test.
Figure 10 is a figure confirming the change in blood glucose by oral glucose tolerance test anti-diabetic activity in the streptozotocin (STZ) type 1 diabetes induction model by centrifugal ethyl acetate- methanol fraction.

Hereinafter, the present invention will be described in detail.

The present invention relates to the use of Centipede grass ( Eremochloa ophiuroides ) provides a composition for preventing and treating diabetes containing the extract as an active ingredient.

The centipedlass extract is preferably, but not exclusively, prepared by a process comprising the steps of:

1) extracting Sentipedgrass with an extraction solvent;

2) cooling the extract of step 1) and filtering; And

3) Concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the centipedegrass of step 1) may be used without limitation such as grown or commercially available. The centipedegrass may be a leaf, a stem, or a root, but leaves are preferably used.

In the above production method, the extraction solvent of step 1) is preferably water, alcohol or a mixture thereof, and the alcohol is preferably C ₁ To C ₂ lower alcohol is preferably used. As the lower alcohol, ethanol or methanol is preferably used, and methanol is more preferably used, but not limited thereto. As the extraction method, it is preferable to use shaking extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added by 2 to 10 times the total amount of the dried centipedgrass, more preferably by 10 times, but not limited thereto. In addition, the extraction temperature is preferably 50 to 120 ° C, more preferably 40 to 45 ° C, but is not limited thereto. The extraction time is preferably 2 to 5 days, but is not limited thereto. In addition, the number of times of extraction is preferably 1 to 3 times, but is not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The reduced pressure concentration is preferably concentrated at 20 ~ 60 ℃ and preferably concentrated at 45 ~ 55 ℃ is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto. The above-mentioned freeze-drying is preferably performed at -50 to -100 ° C, and drying is preferably performed at -70 ° C, but is not limited thereto.

In the above method, the centipedes grass extract is preferably extracted with hot water, but is not limited thereto.

The present invention provides a composition for the prevention and treatment of diabetes containing the organic solvent fraction prepared by extracting the centipedes glass extract with an organic solvent as an active ingredient.

The present invention provides a composition for the prevention and treatment of diabetes comprising the active fraction obtained by separating the fraction of centipedegrass extract by column chromatography as an active ingredient.

The fraction of the centipedlass extract or the active fraction isolated from the fraction is preferably, but not necessarily, prepared by a process comprising the steps of:

1) preparing an organic solvent fraction by adding an organic solvent to the centipede glass extract; And

2) Performing column chromatography on the fraction of step 1) to produce an active fraction.

In the above method, the organic solvent in step 1) is preferably, but not limited to, n-hexane or ethyl acetate.

In the above method, the organic solvent fraction is preferably an ethyl acetate fraction obtained by extracting the centifed glass methanol extract with n-nucleic acid and then removing the soluble layer with ethyl acetate.

In this method, the column chromatography of step 2) is carried out by column chromatography using a filler selected from the group consisting of silica gel, Sephadex, RP-18, polyamide, Toyopearl and XAD resin to obtain an active fraction Can be separated and purified. The column chromatography can be carried out several times by selecting an appropriate filler as necessary.

In the above method, the ethyl acetate fraction of the centifed glass extract of step 2) is additionally adsorbed to column chromatography, and methanol is used as elution solvent. desirable. Specifically, we adsorbed the ethyl acetate fraction of centifedgrass to Flex-Colum ^™ (3 cm × 25 cm) and Toyoperarl HW-40 (75 μm) column chromatography and adsorbed 20, 50 or 70%. Using methanol eluting solvent, ethyl acetate-20% methanol, ethyl acetate-50% methanol and ethyl acetate-70% methanol active fractions were obtained.

In order to analyze the fat conversion of glucose from adipocytes by the centifed glass ethyl acetate-methanol fraction, the present inventors investigated the ethyl acetate-50% and 70% methanol fractions in adipocytes using a 3T3-L1 Preadipocyte cell line. As a result of measuring glucose uptake effect, the level of fat accumulation in adipocytes was low in the group treated with 100 ㎍ / ml of ethyl acetate-50% methanol fraction based on the group treated with low concentration of insulin. It was increased by more than 300% (see Fig. 2 and 3), Cetidegrass extract fractions confirmed the hypoglycemic activity in type 2 diabetic mice ( db / db mice) as a result of the centipedigrass ethyl acetate-50% and 70% methanol fraction 10 days treatment group compared to the diabetes-induced group showed a 70-80% hypoglycemic effect, and by stopping the sample processing The blood sugar level of the mice was elevated again (see FIG. 4), and the glucose oral loading test (OGTT) analysis of the centifed glass ethyl acetate-methanol fraction showed that the mice treated with the centipedigrass ethyl acetate-methanol fraction It was confirmed that the sudden increase in blood sugar was reduced by about 25% compared to the comparison group, and after 150 minutes, the degree of blood sugar drop was increased by about 60% compared to the comparison group (see FIGS. 5 and 6). In the glucose oral loading test with different doses of acetate-methanol fractions, the blood glucose level increased sharply after 60 minutes in the group treated with the tifedigrass ethyl acetate-50% methanol fraction at 40% at high concentrations, It was confirmed that at a low concentration is maintained at 60% level (see Fig. 7), centifed glass ethyl acetate-methanol Intraperitoneal tolerance test (IPGTT) analysis of different doses of water showed high and low concentrations of the centipedigrass ethyl acetate-50% methanol fraction in comparison with the control group. After the time, the blood glucose level was increased by about 80% (see Fig. 8). After the intraperitoneal administration of the centifed glass ethyl acetate-methanol fraction, the glucose oral loading test analysis showed that the control group was passed 4 hours and 30 minutes later. Blood glucose was maintained above 400 mg / dL, but the blood sugar of the group treated with the centipedigrass ethyl acetate-50% methanol fraction dropped below 250 mg / dL. This was about 60% higher than the control group (see Fig. 9), and antidiabetic activity was analyzed in Streptozotocin (STZ) -type 1 induction model by the centifed glass ethyl acetate-methanol fraction. As a result, the group treated with the centipedigrass ethyl acetate-methanol fraction can be seen that the blood sugar drops to the same level as the normal group after 4 hours and 30 minutes. However, the control group maintained 150 mg / dL higher than the normal group even after 6 hours, and 6 hours after the administration of glucose, the group treated with the centipedigrath ethylacetate-methanol fraction lowered the blood sugar by 100% compared to the control group ( See FIG. 10).

Therefore, by confirming that the extract extracted from Sentipedgrass, fractions thereof, and active fractions separated from the fractions effectively regulate blood glucose levels, the composition for preventing and treating diabetes, and the functional food for preventing or improving diabetes, and the pharmaceutical for controlling blood sugar drop It can be usefully used as a red blood composition and health food for controlling blood sugar drop.

In the pharmaceutical composition for preventing and treating diabetes of the present invention, the diabetes is preferably type 1 diabetes or type 2 diabetes, but is not limited thereto.

The present invention provides a pharmaceutical composition for controlling blood sugar drop, which contains the centipedegrass extract as an active ingredient.

Since the pharmaceutical composition of the present invention has little toxicity and side effects, it is a composition that can be used safely even when taken for a long time.

The pharmaceutical composition of the present invention may further contain commonly used excipients, disintegrants, sweeteners, lubricants, flavors and the like, and may be formulated into tablets, capsules, powders, granules, suspensions, Syrups, and other liquid preparations.

Specifically, the pharmaceutical compositions of the present invention can be formulated for oral administration, for example, as tablets, troches, lozenges, aqueous or aqueous suspensions, prepared powders or granules, emulsions, hard or soft capsules, It is formulated into elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, celluloses or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, and disintegrants such as stearic acid Magnesium stearate, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained.

In addition, the pharmaceutical composition of the present invention can be administered orally or parenterally, and it is preferable to select subcutaneous injection, intravenous injection, intramuscular injection, or intra-thoracic injection injection method for parenteral administration. In order to formulate the formulation for parenteral administration, the active fraction of the centipede glass extract, fraction or fraction thereof of the present invention is mixed with water with a stabilizer or a buffer to prepare a suspension, which is formulated into a unit dosage form of ampoule or vial.

The dosage of the active ingredient according to the present invention is appropriately selected depending on the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, the age, sex and condition of the patient, and severity of the disease to be treated. The extract of the present invention may be administered to an adult in an amount of 0.0001-500 mg per 1 kg of body weight per day, preferably in an amount of 0.001-100 mg per one kg of body weight.

The present invention relates to the use of Centipede grass ( Eremochloa It provides a functional food for preventing or improving diabetes containing ophiuroides ) extract as an active ingredient.

In addition, the present invention provides a functional food for preventing or improving diabetes containing the organic solvent fraction prepared by extracting the centipedes glass extract with an organic solvent as an active ingredient.

In another aspect, the present invention provides a functional food for preventing or improving diabetes containing the active fraction of the centifed glass extract additionally separated by column chromatography as an active ingredient.

In the health food of the present invention, the diabetes is preferably diabetes, type 1 diabetes or type 2 diabetes, but is not limited thereto.

The present invention relates to the use of Centipede grass ( Eremochloa ophiuroides ) Provides a health food for controlling blood sugar drop containing the extract as an active ingredient.

The health food of the present invention may be added to the centipede glass extract, its fraction or the active fraction isolated from the fraction as it is, or may be used together with other food or food ingredients, and may be suitably used according to a conventional method.

There is no particular limitation on the kind of the food. Examples of the food to which the centipedegrass extract, the fraction thereof, or the active fraction isolated from the fraction can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gum, ice cream , Various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples, Comparative Examples and Preparation Examples.

However, the following Examples, Experimental Examples, Comparative Examples and Preparation Examples illustrate the present invention concretely, and the contents of the present invention are not limited by Examples, Experimental Examples, Comparative Examples and Preparation Examples.

< Example  1> Sentiped Grass  Preparation of extract

<1-1> Sentiped Grass  Preparation of methanol extract

The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. Leaf samples were stored at -80 ° C. 5 kg of Centipede grass leaf was added with 20 L of 100% methanol (MeOH), ground with a blender, immersed at room temperature for 3 days, and then concentrated under reduced pressure at 50 캜 to obtain 210 g of a centipede glass methanol extract.

<1-2> Sentiped Grass  Preparation of ethanol extract

The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. Leaf samples were stored at -80 ° C. 5 kg of centipedegrass leaves were added with 20 L of 100% ethanol (EtOH), ground with a blender, immersed in the liquid at room temperature for 3 days, and concentrated under reduced pressure at 50 DEG C to obtain 203 g of a centipede glass ethanol extract.

<1-3> Sentiped Grass  Preparation of water extract

The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. The collected leaf samples were stored at -8 ° C. 20 ℓ of water was added to 5 ㎏ of centipedegrass, and the mixture was ground with a blender and then immersed at room temperature for 3 days. The extract was concentrated under reduced pressure at 50 캜 to obtain 198 g of centipede water extract.

<1-4> Sentiped Grass Heat number  Preparation of extract

The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. The collected leaf samples were stored at -8 ° C. 40 g of Centipedgrass leaves were added to 40 L of water and soaked at 90 DEG C for at least 6 hours to obtain 87 g of Centipedgrass hydrothermal extract.

< Example  2> Sentiped Grass Fraction  Produce

The centifed glass methanol extract obtained in Example <1-1> was fractionated according to polarity using an organic solvent.

<2-1> n- Hexane Fraction  Produce

First, as shown in FIG. 1, the concentrated concentrate under reduced pressure of 100% MeOH extract of centifed grass was suspended in water, followed by extraction with hexane (N-Hexane), which is a low polar solvent.

<2-2> Ethyl acetate Fraction  Produce

The water layer of hexane extraction of Example <2-1> was again partitioned with ethyl acetate (EtOAc).

<2-3> Water Fraction  Produce

In Example < 2-2 > in the above Example, fractionation with n-hexane and ethyl acetate (EtOAc) was carried out and finally a soluble fraction with water was obtained. The solvent-extracted fractions were concentrated under reduced pressure and dried to obtain an ethyl acetate fraction (21 g).

< Example  3> Sentiped Grass  Production of active fractions

<3-1> ethyl acetate-20% methanol Fraction  Produce

The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum ^TM (3 cm x 25 cm) using 20% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. An active fraction of 9.41 g of ethyl acetate-20% methanol was obtained.

<3-2> ethyl acetate-50% methanol Fraction  Produce

The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum ^TM (3 cm x 25 cm) using 50% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. An active fraction of 5.74 g of ethyl acetate-50% methanol was obtained.

<3-3> ethyl acetate-70% methanol Fraction  Produce

The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum ^TM (3 cm x 25 cm) using 70% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. Ethyl acetate - 1 g of active fraction with 70% methanol was obtained.

<3-4> Ethyl acetate-30% and 70% methanol Fraction  Component analysis

The contents of maysin and its derivatives were analyzed in the components of the fractions of the centifedgrass extract, partial fractions of the ethyl acetate-30% and 70% methanol fractions. Specifically, 20 μl of sample solution was injected into Agilent Technologies 1200 series HPLC and a mobile phase consisting of 1% formic acid (solution A) and 100% methanol (solution B) was used. The mobile phase is eluted with a 50:50 linear gradient at 100: 0 (solution A: B) in a combined step and 30 minutes, followed by a 50:50 linear gradient at 50:50 for 60 minutes. I was. The flow rate was 0.5 ml / min and the eluted compound was detected at a wavelength of 360 nm. As a result, the contents of eluted macine and its derivatives are shown in Table 1 below.

Compound (% of powder mg) 50% 70% Luteolin 0.1938 1.6864 Isoorientin 3.1703 13.9925 Rhamnosylisoorientin (with Orientin) 1.8745 13.6534 Derhamnosylmaysin 20.2102 9.4434 Maysin 14.0661 19.2835 Sum 39.5151 58.0593

< Experimental Example  1> Sentiped Grass  Ethyl acetate-methanol In the fraction  Fat conversion analysis of glucose by fat cells

Adipocyte lines (3T3-L1 Preadipocyte cell line) were used to determine whether ethyl acetate-50% and 70% methanol fractions enhance glucose uptake by adipocytes. Specifically, the fat cell line was purchased from the American Type Culture Collect (ATCC), put the adipocytes (3T3-L1) to 1 × 10 ⁶ in a ⁶ well plate, so that the cells can grow to 90% Incubated for 24 hours in a CO ₂ incubator. Then, the culture medium was discarded and another medium to help differentiation into adipocytes was added to induce differentiation into adipocytes for 48 hours. The composition of the adipocyte differentiation medium was 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 10 nm / ml insulin, 1 μM dexamethasone ( dexamethasone) is a DMEM medium. Then, when 3T3-L1 cells are induced to differentiate into adipocytes, the crude extract of the centipedegrass, a negative control group, ethyl acetate-50 in each adipocytes in a medium containing low concentration of insulin is added. % And 70% methanol fractions were treated. Glucose was introduced into the adipocytes and the accumulated amount of fat produced through the process of glucose metabolism was confirmed through oil-red (Oil Red) -O staining. The oil-red-O staining solution is a reagent that specifically stains only fats in adipocytes, so that glucose can be broken down and biosynthesized, so that the amount of fat produced in each sample can be compared and analyzed. The solution dyed red using iso-propanol, a dye extraction solution, was measured at 490 nm after extraction.

As a result, when the low concentration of insulin was treated, the accumulation of fat in adipocytes increased by about 300% in the group treated with 100 μg / ml of ethyl acetate-50% methanol fraction. . This ethyl acetate-50% methanol fraction increased the adipocytes to enter glucose into the cells (Fig. 2 and 3).

< Experimental Example  2> Sentiped Grass  Ethyl acetate-methanol In the fraction  Hypoglycemic analysis of experimental animals

Cetipedgrass extract fractions were confirmed to have hypoglycemic activity in type 2 diabetic mice ( db / db mice). C57BL / 6 mice were used as a normal group, and C57BLKS / J- db / db mice were used as a diabetic induction group. Both test animals were imported from Japan's SLC Co., Ltd. through the Central Test Animal Co., Ltd., and the age of import was 7 weeks. After the animals were brought into the laboratory of the laboratory, they were allowed to acclimate for one week while maintaining the temperature of 22 ° C. and the humidity of 50%. Since the blood was collected from the tail at the time of over 9 weeks of age, blood glucose was checked, and the test for diabetic activity was performed from 14 weeks of age when the blood glucose was over 350 mg / dL. Groups of blood glucose levels exceeding the standard (350 mg / dL) were randomly mixed and grouped. The normal and induced groups received oral corn oil daily, and the remaining groups received reagents and samples for each treatment group. It was mixed with corn oil and orally administered daily. Oral dosages were set to 200-300 μl per horse. The reagents and samples were administered continuously for 10 days from the start date, and then the sample administration was stopped for 2 days to see if there was any change in mouse blood glucose, and the change in blood glucose was observed when the sample was processed again.

As a result, it was confirmed that the group treated with the centipedigrass ethyl acetate-50% and 70% methanol fraction for 10 days showed a blood glucose lowering effect of 70-80% compared to the diabetic group, and the mouse was stopped by stopping the sample treatment. As the blood sugar level of the rising again, ethyl acetate-50% and 70% methanol fractions were confirmed to have a significant effect on the blood sugar drop (Fig. 4).

< Experimental Example  3> Sentiped Grass  Ethyl acetate-methanol In the fraction  by Glucose  Oral Load Experiment ( Oral Glucose Tolerance test , OGTT ) analysis

The glucose oral loading test is a widely used method for diagnosing diabetes, and is one of methods for diagnosing diabetes by orally administering glucose in a fasting state to check blood glucose. The experimental animals Central Laboratory Animal Co., Ltd. as a C57BLKS / J- db / db mice imported from Japan SLC Inc. centimeters Phedi glass ethyl acetate through the use in this experiment - then with methanol fractions finished diabetes experiments 2 weeks, wherein After two weeks of acclimation, it was used again in OGTT. The progression of the experiment was to fast the mice for 18 hours to maintain an empty stomach, and the samples (50% or 70% fraction extracts; 10 mg / kg body weight) were mixed with corn oil and pretreated by oral administration 30 minutes prior to glucose administration. Glucose was orally administered at a weight of 2 g / kg, and the change in blood glucose was confirmed six times in 30 minutes.

As a result, the mice treated with the centipedigrass ethyl acetate-methanol fraction reduced the sudden increase in blood glucose by about 25% compared with the control group, and after 150 minutes, the blood glucose drop was increased by about 60% compared to the control group. I could confirm it. It was confirmed that the centifed glass ethyl acetate-methanol fractions glycemic control (Figs. 5 and 6).

< Experimental Example  4> Sentiped Grass  Ethyl acetate-methanol Fraction  Dosage Differently  Glucose oral loading experiment Oral Glucose Tolerance test , OGTT )

C57BL / 6 mice were used as a normal group, and C57BLKS / J- db / db mice were used as a diabetic induction group. Both test animals were imported from Japan's SLC Co., Ltd. through the Central Test Animal Co., Ltd., and the age of import was 7 weeks. After the animals were brought into the laboratory of the laboratory, they were allowed to acclimate for one week while maintaining the temperature of 22 ° C. and the humidity of 50%. Before starting the experiment, fasted for 12 hours to maintain an empty stomach. Thirty minutes prior to administration of glucose, samples (clude or 50% or 70% fraction extracts; 1 mg / kg or 10 mg / kg body weight) were mixed with corn oil and pretreated by oral administration. Glucose was orally administered at 2 g / kg body weight, and then the change in blood glucose total 6 times in 30 minutes was confirmed.

As a result, it was confirmed that the group treated with the centipedigrass ethyl acetate-50% methanol fraction maintained a rapid increase in blood glucose after 60 minutes compared with the comparative group at a high concentration of 40% and a low concentration of 60% (FIG. 7). ).

< Experimental Example  5> Sentiped Grass  Ethyl acetate-methanol Fraction  Dosage Differently   Intraperitoneal sugar load test ( Intraperitoneal tolerance test , IPGTT ) analysis

C57BL / 6 mice were used as a normal group, and C57BLKS / J- db / db mice were used as a diabetic induction group. Both test animals were imported from Japan's SLC Co., Ltd. through the Central Test Animal Co., Ltd., and the age of import was 7 weeks. After the animals were brought into the laboratory of the laboratory, they were allowed to acclimate for one week while maintaining the temperature of 22 ° C. and the humidity of 50%. Before starting the experiment, fasted for 12 hours to maintain an empty stomach. Thirty minutes prior to administration of glucose, samples (clude or 50% or 70% fraction extracts; 1 mg / kg or 10 mg / kg body weight) were dissolved in 10 mM KOH and pretreated by intraperitoneal administration. Glucose was intraperitoneally administered at a weight of 2 g / kg, after which a total change in blood glucose was confirmed 6 times every 30 minutes.

As a result, when comparing the group treated with high concentration and low concentration of centipedigrass ethyl acetate-50% methanol fraction with the comparison group, it was confirmed that the blood sugar drop degree is increased by about 80% after 2 hours (Fig. 8).

< Experimental Example  6> Sentiped Grass  Ethyl acetate-methanol The fraction  After intraperitoneal administration Glucose  Oral Load Experiment ( Oral Glucose Tolerance test , OGTT ) analysis

Diabetes-induced mice used C57BLKS / J- db / db mice. The experimental animals were imported from SLC, Japan through the Central Experimental Animal Co., Ltd., and the age was 7 weeks of age. After the animals were brought into the laboratory of the laboratory, they were allowed to acclimate for one week while maintaining the temperature of 22 ° C. and the humidity of 50%. Before starting the experiment, fasted for 12 hours to maintain an empty stomach. Thirty minutes before the administration of glucose, the sample (clude or 50% or 70% fraction extract; 1 mg / kg or 50 mg / kg body weight) was dissolved in 10 mM KOH and pretreated by intraperitoneal administration, and glucose was 2 g / kg body weight. oral administration in kg. The amount to be administered was set to 200 μl each. A total of 9 changes in blood glucose were confirmed for 5 hours.

As a result, after 4 hours and 30 minutes, the blood sugar of the control group was maintained at 400 mg / dL or more, but the blood sugar of the group treated with centipedigrass ethyl acetate-50% methanol fraction dropped to 250 mg / dL or less. This increased about 60% of the level of hypoglycemia compared to the control (Fig. 9).

< Experimental Example  7> Sentiped Grass  Ethyl acetate-methanol In the fraction  Streptozotocin caused by STZ In the Type 1 Diabetes Induction Model Anti-diabetic  Activity analysis

An experiment was conducted to determine whether the centipedes grass extracts had antidiabetic activity in a streptozotocin (STZ) -type 1 induction model using diabetes induced ICR mice. The experimental animals were imported from SLC, Japan through the Central Experimental Animal Co., Ltd., and the age was 7 weeks of age. After the animals were brought into the laboratory of the animal laboratory, a period of acclimation was given for 1 week while maintaining the temperature of 22 ° C and 50% humidity. Type 1 diabetes was induced by lowering insulin secretion by specifically destroying only pancreatic beta cells using STZ. STZ was dissolved in 50 mM citric acid buffer (pH 4.5) to induce type 1 diabetes through intraperitoneal administration, and injected three times at two-day intervals at a weight of 80 mg / kg. Three days later, the blood glucose was checked, and mice with blood glucose greater than 350 mg / dL were randomly mixed and used for the experiment. Before starting the experiment, fasted for 12 hours to maintain an empty stomach. Samples (50% or 70% fraction extracts; 20 mg / kg body weight) were mixed with corn oil and pretreated for oral administration 30 minutes prior to glucose, followed by oral administration of glucose at 2 g / kg body weight. The blood glucose check confirmed the change in blood sugar total 10 times in 6 hours.

As a result, the group treated with the centipedigrass ethyl acetate-methanol fraction can confirm that the blood sugar drops to the same level as the normal group from 4 hours and 30 minutes. However, the control group maintained 150 mg / dL higher than the normal group even after 6 hours, and 6 hours after the administration of glucose, the group treated with the centipidigras ethylacetate-methanol fraction lowered blood glucose by 100% compared to the control group ( 10).

<Preparation Example 1> Preparation of a medicament for preventing and treating diabetes containing centipedegrass extract as an active ingredient

&Lt; Formulation Example 1 > Preparation of pharmaceutical preparation

<1-1> Manufacture of Powder

Example < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract

Lactose 1.5 g

Talc 0.5 g

The above ingredients were mixed and filled in an airtight container to prepare powders.

<1-2> Preparation of tablets

Tablets containing the centipedegrass extract of the present invention as an active ingredient were prepared as shown in Table 3 below.

Constituent Content (parts by weight) Sentipedragas extract 250 Lactose 175.9 Potato starch 180 Colloidal silicic acid 32 10% gelatin solution 5 Potato starch 160 talc 50 Magnesium stearate 5

250 parts by weight of the extract of the present invention, 175.9 parts by weight of lactose, 180 parts by weight of potato starch and 32 parts by weight of colloidal silicic acid. 10% gelatin solution was added to the mixture, followed by pulverization and passed through a 14-mesh sieve. This was dried, to which 160 parts by weight of potato powder, 50 parts by weight of talc and 5 parts by weight of magnesium stearate were added, and the resulting mixture was purified.

<1-3> Preparation of capsules

Example < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract

Corn starch 5 g

4.9 g of carboxycellulose

After mixing the above components to prepare a powder, the powder was filled in a hard capsule according to a conventional preparation method of a capsule to prepare a capsule.

&Lt; 1-4 > Preparation of injection

Example < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract

Sterile sterilized water for injection

pH adjuster

(2 ml) per ampoule according to the usual injection preparation method.

<1-5> Manufacture of liquid agent

Example < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract

10 g per isomer

5 g mannitol

Purified water quantity

Each component was dissolved in purified water in accordance with a conventional method for producing a liquid agent, and the lemon flavor was added in an appropriate amount, followed by mixing the above components. Then, purified water was added to adjust the total amount to 100, and the solution was filled in a brown bottle and sterilized to prepare a liquid preparation.

&Lt; 1-6 > Preparation of syrup preparation

Syrup containing the centipedegrass extract of the present invention as an active ingredient was prepared as shown in Table 2.

Constituent Content (parts by weight) Sentipedragas extract 2 saccharin 0.8 Party 25.4 glycerin 8 Spices 0.04 ethanol 4 Sorbic acid 0.4 Distilled water 60

Preparation Example 2 Preparation of Health Foods Containing Centipedgrass Extract as Active Ingredient

<2-1> Production of flour food

0.5-5.0 parts by weight of the centipedglass water extract of Example <1-3> was added to wheat flour, and the mixture was used to prepare bread, cake, cookies, crackers and noodles.

<2-2> Production of soups and gravies

0.1 to 5.0 parts by weight of the centipedglass water extract of Example <1-3> was added to the soup and the juice to prepare a health promotion meat product, noodle soup and juice.

<2-3> Preparation of ground beef

10 parts by weight of the centipede water extract of Example <1-3> was added to the ground beef to prepare ground beef for health promotion.

<2-4> Production of Dairy Products

5-10 parts by weight of the centipedglass water extract of Example <1-3> was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

&Lt; 2-5 >

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The centipede water extract of Example <1-3> was concentrated under reduced pressure in a vacuum concentrator, dried by spraying and dried in a hot air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The above-prepared cereals, seeds and centipede water extract of Example <1-3> were prepared in the following proportions.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

The extract (3 parts by weight) of Example <1-3>,

(0.5 part by weight),

(0.5 parts by weight)

Preparation Example 3 Preparation of Functional Drink Containing Centipedgrass Extract as Active Ingredient

<3-1> Production of health drinks

(5%) of the centipede glass extract of the present invention was homogeneously mixed with a mixture of liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5% And packaged in small containers such as glass bottles and plastic bottles.

<3-2> Preparation of vegetable juice

Vegetable juice was prepared by adding 5 g of centifedgrass extract of the present invention to 1,000 ml of tomato or carrot juice.

<3-3> Production of fruit juice

1 g of centifedgrass extract of the present invention was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

Claims (14)

Centipede grass (Centipede grass, Eremochloa ophiuroides ) water, C ₁ to C ₂ low alcohol or a mixed solvent extract thereof comprising as an active ingredient a pharmaceutical composition for preventing and treating diabetes.
delete The pharmaceutical composition for preventing and treating diabetes of claim 1, wherein the lower alcohol is ethanol or methanol.
[Claim _2] The pharmaceutical composition for preventing and treating diabetes of claim 1, wherein the water of Centipedgrass, C ₁ to C ₂ lower alcohol, or a mixed solvent extract thereof is extracted with hot water.
A pharmaceutical composition for preventing and treating diabetes comprising the fraction prepared by additionally extracting centipedes grass extract with n-hexane or ethyl acetate as an active ingredient.
delete 6. The pharmaceutical composition for preventing and treating diabetes of claim 5, wherein the fraction is an ethyl acetate fraction obtained by extracting the centifed glass methanol extract with n-nucleic acid and removing the soluble layer with ethyl acetate.
6. The pharmaceutical composition for preventing and treating diabetes of claim 5, wherein the fraction comprises an active fraction obtained by additionally extracting the ethyl acetate fraction of the centipedegrass methanol extract with methanol as an active ingredient.
9. The pharmaceutical composition for preventing and treating diabetes of claim 8, wherein the methanol is 20% to 70% by weight methanol in the total aqueous solution.
10. The pharmaceutical according to any one of claims 1, 3 to 5 or 7 to 9, wherein the diabetes is type 1 diabetes or type 2 diabetes. Composition.
A functional food for preventing and ameliorating diabetes, comprising the water of sentigraphgrass of claim ₁ , a lower alcohol of C ₁ to C ₂ or a mixed solvent extract thereof, a fraction of claim 5, or an active fraction of claim 8 as an active ingredient.
12. The functional food for preventing and improving diabetes of claim 11, wherein the diabetes is type 1 diabetes or type 2 diabetes.
A pharmaceutical composition for controlling blood sugar drop, comprising the water of Centipegrass of claim ₁ , a lower alcohol of C ₁ to C ₂ or a mixed solvent extract thereof, a fraction of claim 5 or an active fraction of claim 8 as an active ingredient.
Claim 1 of the sentimental glass of water, C ₁ to C ₂ lower alcohol or a mixed solvent extract thereof, a fraction of claim 5 or an active fraction of claim 8 as an active ingredient for controlling blood sugar drop.

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