KR20130055395A - Composition for inhibiting ptp1b containing prunellae, artemisia extracts - Google Patents

Abstract

PURPOSE: A mixture of a Prunella vulgaris extract and an Artemisia argyi herb extract is provided to suppress PTP1B(protein tyrosine phosphatase B1), and to prevent or treat diabetes. CONSTITUTION: A composition for suppressing PTP1B contains a Prunella vulgaris extract and/or an Artemisia argyi herb extract in a ratio of 1:1. A drug composition contains the composition for suppressing PTP1B as an active ingredient. A health food composition contains the composition for suppressing PTP1B as an active ingredient. A method for preparing the composition for suppressing PTP1B comprises: a step of extracting Prunella vulgaris and Artemisia argyi herb with water and filtering; a step of concentrating the liquid; and a step of drying the concentrate. [Reference numerals] (AA,EE) Control group; (BB,GG) Artemisia argyi herb; (CC,FF) Prunella vulgaris; (DD,HH) Mixture of Prunella vulgaris and Artemisia argyi herb

Description

Composition for inhibiting protein tyrosine dephosphorylase enzyme comprising haejucho, leafy leaf extract as an active ingredient {COMPOSITION FOR INHIBITING PTP1B CONTAINING PRUNELLAE, ARTEMISIA EXTRACTS}

The present invention relates to a composition comprising the edible fruit, leaf extract as an active ingredient, and more specifically, the main symptoms of diabetes by inhibiting protein tyrosine dephosphatase B1 (hereinafter referred to as PTPB1) from the edible fruit, leaf extract The present invention relates to a substance having a hypoglycemic effect on phosphorus hyperglycemia, a method of extracting the same, and a pharmaceutical composition for preventing and treating diabetes and a hypoglycemic agent comprising the same as an active ingredient, and a health food composition.

Diabetes mellitus is a disease in which glucose is not absorbed into the blood cells, resulting in high blood sugar, which causes glucose to escape into the urine. After eating, glucose is not absorbed into the cells, resulting in an imbalance in energy metabolism. At the same time, because a large amount of water is urinated, water intake increases, causing various complications such as abnormal kidney function, or excessive blood sugar increase, which is a metabolic disease that causes life by shock or coma. Diabetes is a type 1 diabetes (insulin-dependent) caused by the lack of insulin in the pancreas and insulin, but type 2 diabetes occurs due to decreased insulin activity or weakened sensitivity in peripheral tissues resulting in insulin resistance resulting in problems with blood sugar control. It is divided into diabetes (insulin-independent). Type 1 diabetes is called insulin-dependent diabetes because it controls blood sugar when injected with insulin, and is often congenital. On the other hand, type 2 diabetes is called insulin-independent diabetes because it does not control blood sugar by insulin injection, but because blood sugar is controlled by a hypoglycemic agent or diet, 95% or more of the diabetic patients. Recently, serious obesity has been caused by changes in diet and lack of exercise, and type 2 diabetes patients are increasing. Most of the diabetic patients in Korea are type 2 diabetic patients, and if diabetes persists for a long time, blood lipid concentrations increase in addition to the increase in blood sugar, which leads to complications such as cardiovascular diseases such as fatty liver, arteriosclerosis, and coronary heart disease, diabetic kidney and retinal diseases. It is urgent to develop a treatment for it.

Currently, insulin resistance is recognized as the most important pathological mechanism of type 2 diabetes, which means that even if the insulin is sufficient in the body, sugar is not properly utilized, and various blood glucose lowering agents are used to treat it. Glucose enhancers can be divided into sulfonylurea, biguanide, and alpha-glucosidase inhibitors, depending on the mechanism of action and the point of action of the drug. Thiazolidinedione (TZD) -based drugs that activate (PPAR-γ, Peroxisome Proliferator activated receptor γ) are also being developed. However, the oral hypoglycemic agent alone does not drop blood sugar effectively, and most drugs have side effects. Therefore, it is urgent to develop a safer diabetes drug, and a drug made of a natural extract alleviates these side effects and supplements the efficacy of drugs recently. Is getting.

Recently, as association between type 2 diabetes and PTPB1 has been revealed, its inhibitor has emerged as a therapeutic agent for diabetes. In mice with the PTPB1 gene removed, insulin resistance was eliminated and type 2 diabetes was not induced or obesity was suppressed. Also, PTPB1 inhibitors were found to be resistant to blood sugar and overcome insulin resistance in rats with type 2 diabetes. It has been reported to be effective against type 2 diabetes. (J. Med. Chem. 42, 3199-3202, 1999; J. Biol. Chem. 275, 10300- 10307, 2000)

Prunellae is a perennial herb that belongs to Purnellae Herba and is the ground part of Lamiaceae, Dume Lamiaceae, and Lamiaceae. It is used as a Chinese medicine in China, and it is widely used in Chinese medicine and folk medicine as a medicine for pulmonary tuberculosis, jaundice, high blood pressure, and uteritis. It contains prunellin, ursolic acid, caffein acid, rutin, hyuperoside and water-soluble inorganic salts as its ingredients, and it is known to have diuretic, astringent, and hemostatic effects. However (Korean Patent Registration 849611) is not known to be effective in diabetes.

Artemisia argyi Herb is a plant belonging to the family Asteraceae (Compositae) and has been used a lot as a herbal medicine since ancient times. Dongbogam describes that there are oriental pharmacological effects such as warm hemostasis, acid dehumidification, sweat dehumidification, diarrhea, landscaping, ophthalmology and hemostasis. Isocoumarin, coumarin, and diterpentaxone are known chemical compounds of the young leaf, and eupatilin, a flavonoid component, is known as an antiallergic effect (Japanese Laid-Open Patent Publication No. 59-155314). Although it has been found to be useful as a disease treatment agent (Korean Patent Registration 127777), it is not known to be effective in diabetes.

Recently, the invention of the anticancer effect of haegocho is actively progressing (Korean Patent Registration 849611), but it is not known that it is effective in diabetes. Eupatilin (eupatilin), a component of the lobe, has been known as an anti-allergic effect (Japanese Patent Publication No. 59-155314) and has recently been shown to be useful as a gastrointestinal disease treatment agent in Korea (Korean Patent Registration 127777). Yes is not known.

It has been reported that PTPB1 inhibitors have an effect on type 2 diabetes, such as hyperglycemia and overcoming insulin resistance in type 2 diabetes model mice. (J. Med. Chem. 42, 3199-3202, 1999; J. Biol. Chem. 275, 10300- 10307, 2000)

An object of the present invention to inhibit the enzyme activity of PTP1B to prevent and treat diabetes, hypoglycemic effect, haejucho, leaf extract, pharmaceutical composition using the extract as an active ingredient, health food composition and its manufacturing method do.

The present invention has a composition for inhibiting PTP1B obtained from hachichos and leaf extracts. Preferably, the vinegar extract and leaf extract are in a ratio of 1: 1, which is effective in preventing and treating diabetes and lowering blood sugar. Moreover, the composition is a pharmaceutical composition and a health food composition which use this composition as an active ingredient. An extraction filtration step of preparing and extracting the composition for inhibiting PTP1B, adding water to at least one pulverized product of vinegar and leaflets; Concentrating step of concentrating the filtrate obtained in the extraction filtration step; And a method for producing a composition for inhibiting PTP1B consisting of a drying step of drying the concentrate obtained in the concentration step, preferably the drying step is a configuration of the present invention that the spray drying method.

Ingestion of haejucho, leafy mixed extract can increase the effect of insulin and delay the absorption of glucose by preventing PTP1B enzyme involved in dephosphorylation during insulin signaling of body tissues, thereby preventing or improving diabetes. . Therefore, the extract obtained from the edible vinegar of the present invention, as a natural extract, because it has the effect of lowering blood sugar, preventing and treating diabetes as a natural extract, it can be used as a herbal medicine, health food, etc. containing it as an active ingredient.

1 to 6 show the manufacturing process of the hawaii and leaf extract powder, corresponding to Examples 1 to 6,
Figure 7 is a result and graph of the PTP1B inhibitory activity of the extract of Hagocho, leaf leaf,
FIG. 8 is a graph of blood glucose levels measured when oral administration of hyphae and lobules to mice induced with diabetes with streptozotocin. FIG.

The present invention relates to a health food or pharmaceutical composition effective in the prevention and treatment of diabetes, hypoglycemic composition consisting of hachicho, leaf extract. The mixing ratio of the edible fruit and leaf extract of the present invention is preferably 1: 1, and the extraction, filtration, concentration, and drying methods may be used with conventional methods. It is particularly preferable that the drying step is spray drying. Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the examples.

Example  One And seconds , Leaf extract powder  Manufacture process ( 1 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. The vinegar was purchased from Dong-Eui Herbal Herbs Co., Ltd. as a dry medicinal herb, and extracted with 20 times of water, extracted at 95 ° C. for 2 hours, filtered using a filter cloth and a mesh, and concentrated at 60 ° C. using a vacuum concentrator. The concentrate was spray dried using a spray dryer to obtain an extract. The leaf extracts were also obtained in the same manner as in the preparation of the extract of Hachocho extract. The upper leaf extract powder and leaf extract extract powder 1: 1.

Example  2 And seconds , Leaf extract powder  Manufacture process( Figure 2 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. The vinegar was purchased from Dong-Eui Herbal Herbs Co., Ltd. as a dry medicinal herb, and extracted with 20 times of water, extracted at 95 ° C. for 2 hours, filtered using a filter cloth and a mesh, and concentrated at 60 ° C. using a vacuum concentrator. The leaf extracts were also obtained in the same manner as in the preparation of the extract of Hachocho extract. The herbaceous leaf extract and leaf extract were mixed 1: 1 to be spray dried using a spray dryer to powder.

Example  3 And seconds , Leaf extract powder  Manufacture process( 3 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. The vinegar was purchased from Dong-Eui Herbal Herbs Co., Ltd. as a dry medicinal herb, extracted with 2 times the water and extracted at 95 ° C. for 2 hours, and then filtered using a filter cloth and a mesh. The leaf extracts were also obtained in the same manner as in the preparation of the extract of Hachocho extract. The herbaceous leaf extract and leaf extract were mixed 1: 1 and concentrated at 60 ° C. with a reduced pressure concentrator, and then spray dried using a spray dryer to powder.

Example  4 And seconds , Leaf extract powder  Manufacture process( Figure 4 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. The vinegar and young leaf were purchased from Dong-Eui Herbal Medicine Co., Ltd. as a dry medicinal herb, mixed 1: 1, and ultrasonically extracted at 95 ° C. for 2 hours by adding 20-fold water, followed by filtration using a filter cloth and a mesh. The filtrate was concentrated at 60 ° C. with a vacuum condenser, and then spray dried using a spray dryer to powder.

Example  5 And seconds , Leaf extract powder  Manufacture process( 5 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. The vinegar and leafy leaves were purchased from Dong-Eui Herbal Herbs Co., Ltd. as a dry herb, pulverized and mixed 1: 1. 20 times of water was added thereto, followed by stirring extraction at 95 ° C. for 2 hours, followed by filtration using a filter cloth and a mesh. The filtrate was concentrated at 60 ° C. with a vacuum condenser, and then spray dried using a spray dryer to powder.

Example  6 And seconds , Leaf extract powder  Manufacture process( Figure 6 )

In order to obtain an extract having a PTP1B inhibitory effect was carried out as follows. Hagicho and young leaf were purchased from Dongeui Oriental Herbal Medicine Co., Ltd. as a dry herb, and mixed 1: 1. 20 times of water was added thereto, followed by stirring extraction at 95 ° C. for 2 hours, followed by filtration using a filter cloth and a mesh. The filtrate was concentrated at 60 ° C. with a vacuum condenser, and then spray dried using a spray dryer to powder.

Test Example  1 of the extract PTP1B  Measurement of inhibitory activity

After immobilizing the substrate to be dephosphorylated by Phosphatase reaction on the protein chip, the natural material was evaluated by measuring the degree of dephosphorylation of the substrate by adding buffer solution and phospatase necessary for the enzyme reaction.The PTP1B protein chip was manufactured by Proteogen Co., Ltd. It was purchased and used.

In order to induce self phosphorylation of the substrate, ATP 100uM was treated and reacted at 30 ° C. for 1 hour, followed by overnight incubation at 4 ° C. to fix the substrate on the chip. Washing the protein chip with 0.5% tween in PBS buffer and blocking the reaction with 3% BSA for 1 hour to prevent non-specific reaction, and then treated each sample and treated with PTP1B at a concentration of 1ng / ul to induce dephosphorylation. In order to determine whether PTP1B is dephosphorylated, a phosphate-specific antibody is treated and reacted for 1 hour, and then a secondary antibody (anti-IgG-Cy5) capable of detecting the antibody is reacted to react with PTP1B. Activity was measured. The results are shown in FIG.

Na ₃ VO ₄ used as a control showed 70.68% of inhibitory activity, whereas the results of 84.04% and 85.67% of inhibitory activity were measured, respectively. PTP1B inhibitory activity was shown.

Test Example  2 By streptozotocin administration  Diabetes Induction and Blood Sugar Identification Test

Four-week-old male C57BL7 mice were supplied from Korean Biolink, followed by one week of laboratory purification, and five-week-old mice were used for the experiment.

To induce diabetes in the mice, streptozotocin was dissolved in 0.1 M citrate buffer (pH4.5) and administered intraperitoneally at a concentration of 60 mg / kg, followed by second administration of streptozonosine in the same manner after 3 days of curing. Pleurotus eryngii and lobe were orally administered twice a day at a concentration of 100 mg / kg. Blood glucose was measured from blood shed in the tail vein of the mouse at 3 days intervals after the second streptozotocin administration. The results are shown in FIG.

Claims (7)

Composition for inhibiting protein tyrosine dephosphatase 1B (PTP1B), characterized in that consisting of one or more of the vinegar extract and leaf extract.
The composition for inhibiting PTP1B according to claim 1, wherein the edible extract and the leaf extract are mixed in a ratio of 1: 1.
The composition for inhibiting PTP1B according to claim 1 or 2, wherein the composition for inhibiting PTP1B is effective in lowering blood sugar, preventing and treating diabetes.
A pharmaceutical composition containing the composition for inhibiting PTP1B according to any one of claims 1 to 2 as an active ingredient.
A health food composition containing the composition for inhibiting PTP1B according to any one of claims 1 to 2 as an active ingredient.
An extraction filtration step of extracting and filtering water by adding water to at least one pulverized leaf of the vinegar and the leaves;
Concentrating step of concentrating the filtrate obtained in the extraction filtration step; And
Method for producing a composition for inhibiting PTP1B, characterized in that consisting of a drying step of drying the concentrate obtained in the concentration step.
According to claim 6, The drying step is a method for producing a composition for inhibiting PTP1B, characterized in that the spray drying method.

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