KR101336068B1 - Anti-diabetes composition comprising oriental herbal extracts and fractions - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of obesity or diabetes comprising a fraction obtained by extracting or fractionating an alcohol extract having 1 to 4 carbon atoms of the composition containing ephedra, bale, rhubarb as an organic solvent. The composition is excellent in weight loss and body fat reduction effect without the toxicity in vivo, and has the effect of inhibiting insulin resistance, and medicines for the prevention and treatment of obesity or diabetes and health functional food for the prevention and improvement of obesity or diabetes Can be used as

Description

Anti-diabetes composition comprising oriental herbal extracts and fractions}

The present invention relates to a composition for the prevention and treatment of diabetes comprising the fraction obtained by extracting or fractionating the extract of the composition comprising ephedra, bale, rhubarb with an organic solvent as an active ingredient.

There are many changes in eating habits due to recent economic growth and lifestyle changes. In particular, busy modern people are gaining weight and obesity due to high calorie diets such as fast food and low exercise. Overweight refers to body mass index (BMI, obesity index divided by the square of body weight) between 25 and 30, and obesity is associated with body mass index> 30.

Two-thirds of US adults are overweight or obese, and overweight increases blood pressure and cholesterol levels, increasing the incidence of various adult diseases, including heart disease. The effects of obesity on various diseases are so severe that it is known that 80% of diabetics worldwide and 21% of heart disease are caused by obesity. In addition, the incidence of cervical cancer, kidney cancer, breast cancer, such as various types of cancer are obesity and associated, directly or indirectly, and the case of obese or overweight sleep apnea, osteoarthritis, gallstones also higher than normal (Stein CJ, Colditz GA. The epidemic of obesity. J Clin Endocrinol Metab 2004; 89: 2522-2525; Kopelman PG, Obesity as a medical problem, Nature 2000 Apr 6; 404 (6778): 635-43.).

It is difficult to treat obesity in any one way because it is caused by a combination of various factors such as genetic, environmental, social, and mental factors, rather than a disease caused by one cause. Current methods of treating obesity include diet, exercise, and behavioral therapy, as well as methods of correcting lifestyle, drug therapy, and surgical treatment. Although aggressive efforts should be made to correct lifestyle habits before medication or surgical treatment, it is not easy to correct lifestyle habits, and there is a limit to the weight that can be reduced by lifestyle correction alone. Therefore, in many cases, lifestyle correction and medication are necessary.

Although many anti-obesity agents are being developed every year for the treatment of obesity, there are not many obesity agents available at present, and most of them are limited to digestive or appetite-suppressing agents. In the case of digestive or appetite control agents, they are classified as psychotropic drugs because of habit, and digestion inhibitors show side effects such as diarrhea and constipation. By 2010, the US FDA approved long-term obesity treatment drugs include sibutramine (Reductil), which suppresses the reuptake of norepinephrine and serotonin, and lipase, which is secreted by the pancreas and digestive system (orlistat; Xenical), which has the effect of inhibiting lipase (Yanovski SZ, Yanovski JA, Obesity, N Engl J Med 2002; 346: 591-602).

However, sibutramine has been withdrawn in the US and Korea in October 2010 due to the risk of cardiovascular events such as myocardial infarction and stroke, which are common with side effects such as elevated blood pressure, insomnia, dry mouth, and dizziness. In addition, Orlistat has been reported to have side effects such as diarrhea, fatty liver, loss of gold, etc. In the case of low fat intake compared to Westerners, the effect of the drug is not clear and its use is limited (Kim, Journal of Obesity, 1998; 7 (4): 287-92.).

Therefore, there is a need for an anti-obesity preventive and therapeutic agent that can replace the obesity treatment drug, which has yet to be effective in inhibiting obesity and has been confirmed for long-term safety.

On the other hand, diabetes is a disease in which glucose present in the blood is discharged through urine, and is one of chronic degenerative diseases that does not cure fundamentally. Due to changes in diet and lack of exercise, a major change has occurred in the energy metabolism of the human body, and chronic degenerative diseases such as diabetes are increasing. In Korea, the prevalence of diabetes is known to reach 5-10% and shows a continuous increase. In the United States, diabetes has increased six-fold over the last 40 years, and this increase is expected to reach 26 million patients by 2050.

The etiology of type 2 (insulin-independent) diabetes, which accounts for more than 95% of diabetes, is known to be a combined disorder of two causes: insulin secretion disorder and insulin resistance. Diabetes mellitus is a chronic hyperglycemia due to this complex disorder.

Insulin secretion disorder refers to a situation in which an appropriate amount of insulin is not secreted from the beta cells of the pancreas according to the blood glucose level, which includes both a quantitative decrease in insulin secreting beta cells and a functional distribution disorder. Insulin resistance is a condition in which secreted insulin reaches the target organs in the bloodstream, but the insulin action and sensitivity of the target cells are lowered. It is generally considered to be a signal transduction disorder after cell membrane receptor binding, , Obesity, decreased physical activity, and hyperglycemia or dyslipidemia. Insulin resistance requires higher levels of insulin to overcome resistance, and conversely, hyperglycemia caused by insufficient insulin secretion may worsen insulin resistance again.

Currently, the treatment of insulin-independent diabetes mellitus, diet therapy, exercise therapy, sulfonurea drug, biguanide drug, α-glucosidase inhibitor, insulin, etc. are used. Linidin- or thiazolidinidione-based drugs, insulins with various action times have been developed and applied in clinical practice. However, these therapeutic agents have a problem of causing low side effects or many side effects such as liver dysfunction, hypoglycemia, lactic acidosis and the like. Therefore, it is necessary to reduce the side effects, improve the metabolic disorders, and the like, and to develop an excellent diabetes treatment agent for long-term use.

Herba Ephedrae is an anti-inflammatory and antipyretic analgesic effect, which acts as a dual action of hypertension and hypertension, hypertension and elevation of blood pressure, diuretic action, It has been known to have anti-inflammatory, anti-inflammatory and anti-allergic properties, and is known to have anti-inflammatory properties such as cold and upper respiratory infections, pneumonia, bronchitis, bronchial asthma, pertussis, hypotension, heart disease, mucocutaneous disease, nephritis and edema, . In addition, as a constituent drug of mah-hong-tang, it is used when there is a headache due to a cold of sunbath (solar disease), fever, but it does not like to breathe and does not sweat, and arthralgia accompanied by fever, back pain, acute period of rheumatism It is used for asthma (王 本 祥 主 編. 现代 中药 药理 與 clinical. 天津: 天津 科技 飜 譯 Publishing Co., 2004, ISBN 7-5433-1801-6, 1448-1460 .; 王 浴 生, 武 文 龍 · ュ 春春生 主 編. 中 薬 药理 应用(Second Edition), Beijing: People's Health Publications, 2000, ISBN 7-117-03057-7 / R. 3058, 1105-1119.).

Thallus Laminariae seu Eckloniae is a medicinal herb that has a thyroid function improving action, a hypotensive action, an anticoagulant action, a cardiovascular action, a lipid improvement action, a prevention and inhibition action of arteriosclerosis, an anticancer action, a thyroid gland, hypertension, diabetes, It is used for diseases, epidemic hemorrhagic fever, cataracts, dry cough (dry cough, dry cough) (Wang Xiang Xiang, Hyundai Xicheng Pharmaceuticals and Clinics, Tianjin: Tianjin Science and Technology Publishing Co., 2004, ISBN 7-5433-1801-6, 910-914. ; 王 浴 生, 武 文 龍 · 春春生 主 編. 中药 药理 应用 (2nd edition), Beijing: People's health publication, 2000, ISBN 7-117-03057-7 / R. 3058, 684-688.).

Radix et Rhizoma Rhei is an antioxidant that is used for the treatment of diarrhea, liver protection, pancreatic secretion, bile secretion, antimicrobial action, antiviral action, antipyretic and anti-inflammatory action, There is a double regulatory action of metabolism improvement, hemostasis, and immune function. Acute infectious fever and constipation, epidemic hemorrhagic fever, rapid growth obstruction, various acute pesticide poisoning, acute pancreatitis high fever, acute cholecystitis, upper gastrointestinal bleeding, It is used for necrotizing enterocolitis, diabetic nephropathy, dyspepsia and constipation, chronic gastritis and peptic ulcer, hyperlipidemia, pregnancy hypertension (王 本 祥 主 編. 现代 中药 药理 與 clinical. 天津: 天津 科技 飜 譯 clinic, 2004, ISBN 7-5433 Beijing, People's Health Publishing Company, 2000, ISBN 7-117-03057-7 / R. 3058, No. 1, pp. -1801-6, 115-123. 70-80.).

The inventors of the present invention have found that, in the previous studies, mahwang, baguette and rhubarb are each slightly effective in suppressing obesity, but when the mahwang, wrapping and rhubarb are mixed at a certain ratio, the respective substances act synergistically, , Which is a more effective inhibitor of obesity. In addition to the synergistic action, the bale minimizes adverse effects of the ephedrine and rhubarb, such as ephedrine alkaloid, which is a major component of epidrine, or adenosine, which is a major component of rhubarb . Wherein the composition comprising mahwang, bales and rhubarb has a weight ratio of 10% to 50%: 20% to 50%: 20% to 50% of mahwang, Side effects and the like (Patent Registration No. 10-0888068).

As described above, due to the limitations of drugs developed for the treatment of obesity, there is an urgent need for development of drugs with a high mechanism of suppressing obesity and having new side effects with low side effects. Therefore, the present inventors believe that herbal medicine may be an alternative. I thought and studied. Continuing studies with ephedra, bales and rhubarb, the fractions of the ethanol extract of the composition were significantly more effective in inhibiting obesity, such as preventing weight gain, without liver toxicity at a low dose of 1/5 compared to hot water extracts. The present invention was completed by confirming that it has an effect of inhibiting insulin resistance and is effective in treating diabetes due to insulin resistance.

An object of the present invention is to provide a composition for the prevention and treatment of obesity or diabetes comprising the fraction obtained by extracting or fractionating the ethanol extract of the composition comprising ephedra, bale and rhubarb (GGEx18) with an organic solvent.

Another object of the present invention is a health function for the prevention and improvement of obesity or diabetes comprising the fraction obtained by extracting or fractionating the ethanol extract of the composition containing ephedra, bales and rhubarb (GGEx18) with an organic solvent as an active ingredient To provide food.

In order to achieve the above object, the present invention relates to a composition comprising a composition comprising 10% to 50%: 20% to 50%: 20% to 50% by weight of a raw material herb, A pharmaceutical composition for the prevention and treatment of obesity comprising an extract obtained by extracting Kang Ji-Hwang 18 (GGEx18) with an alcohol having 1 to 4 carbon atoms and then extracting or fractionating with an organic solvent as an active ingredient.

The present invention also provides a health functional food for the prevention and improvement of obesity or diabetes, including a pharmaceutical composition for the prevention and treatment of obesity or diabetes, including a pharmaceutically acceptable carrier, and a food supplement. do.

Hereinafter, the present invention will be described in more detail.

The technical terms and scientific terms used in the present invention can be construed as meaning ordinary meanings understood by those of ordinary skill in the art without departing from the scope of the present invention.

In the present invention, a composition comprising 10% to 50% of 20% to 50% of 20% to 50% of mahwang, wrapping and rhubarb is referred to as Gyeongsanggang Jangjwan 18 (GGEx18).

The composition according to the present invention can be obtained as follows. GGEx18 is pulverized and placed in an Erlenmeyer flask equipped with a condenser and extracted with a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. More preferably, 70% ethanol is used. The extraction method can be impregnated or warmed at room temperature, more preferably by hot water extraction (1.5 L x 6) for 2 hours. The extract is filtered and concentrated using a rotary vacuum evaporator. The ethanol concentrate was successively fractionated with dichloromethane, ethyl acetate and butanol, and then treated with sodium sulfate and filtered to concentrate each fraction. The dichloromethane fraction (CF), ethyl acetate fraction (EF) and ethyl acetate fraction (IF), butanol fraction (BF) and water fraction (HF) except ethyl acetate and water layer are obtained.

The composition of the present invention may include one or more known active ingredients having a prophylactic and therapeutic effect of obesity or diabetes, together with the fraction fractionated with an organic solvent from an ethanol extract of GGEx18.

The present invention may be provided as a pharmaceutical composition further comprising suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions.

The composition of the present invention can be administered orally or parenterally at the time of clinical administration and can be used in the form of a general pharmaceutical preparation. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrating agent and a surfactant is usually used. Solid preparations for oral administration are prepared by mixing tablets, pills, powders, granules and capsules. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances and preservatives may be included in addition to water and liquid paraffin, which are commonly used simple diluents. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and gelatin can be used.

The dosage unit may, for example, comprise 1, 2, 3 or 4 times the individual dose or may comprise 1/2, 1/3 or 1/4 times the individual dose. The individual dosages preferably include the amount in which the active drug is administered in one dose, which usually corresponds to the full, half, 2/3 or 1/4 times the daily dose. The effective dose of the composition of the present invention is 100 to 300 mg / kg, preferably 140 to 270 mg / kg, and 1 to 6 times a day May be administered.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of obesity or diabetes.

The composition of the present invention can be added to a dietary supplement for the purpose of preventing and improving obesity or diabetes. When the composition of the present invention is used as a food additive, the composition can be added as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the composition of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like . The ratio of the natural carbohydrate is generally about 0.01 to 0.4 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The composition of the present invention has no toxicity or side effects in vivo, is excellent in safety, and is excellent in the effect of reducing body weight and reducing body fat. Thus, it can be used as a medicine for prevention and treatment of obesity and a health functional food for prevention and improvement of obesity. In addition, by reducing the insulin resistance to reduce the blood glucose and insulin when diabetes is excellent, it can be used as a medicine for the prevention and treatment of diabetes and health functional food for the prevention and improvement of diabetes.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a separation diagram of a composition comprising mahwang, wrapping, and rhubarb.
FIG. 2 is a graph showing changes in body weight change in an animal model during the experimental period.
FIG. 3 is a graph showing weight gain observed in an animal model during an experimental period. FIG.
FIG. 4 is a graph showing a change in the efficiency of the mouse diet. FIG.
FIG. 5 is a view showing the weight of mouse adipose tissue. FIG.
FIG. 6 is a photograph of mouse adipose tissue after hematocylin-eosin staining. FIG.
FIG. 7 is a graph comparing sizes of adipocytes in the mouse adipose tissue of FIG. 6. FIG.
Fig. 8 is a photograph of mouse liver tissue stained with hematocylin-eosin staining. Fig.
FIG. 9 is a graph comparing damage levels in mouse liver tissue. FIG.
10 is a graph comparing the concentrations of AST and ALT in mouse serum.
11 is a diagram comparing the concentration of glucose in mouse serum.
12 is a graph comparing the concentration of insulin in mouse serum.

Hereinafter, preferred embodiments, experimental examples and production examples are provided to facilitate understanding of the present invention. However, the following Examples, Experimental Examples and Preparation Examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the Examples, Experimental Examples and Preparation Examples.

Example  1: Preparation of Experimental Material

① The composition of Gyeongsin Kang Ji-Hwan 18 (GGEx18) was purchased from Busan, South Korea (Composition ratio: 40% for mahwang, 40% for packing, 20% for rhubarb) .

② 1 kg of ground GGEx18 was placed in a Erlenmeyer flask equipped with a cooler, and 1.5 L of 70% ethanol (EtOH) was added, followed by hot extraction (1.5 L × 6) for 2 hours.

(3) The extract was filtered and concentrated using a rotary vacuum evaporator.

④ The 70% ethanol concentrate (306.48 g) was dissolved in water (H ₂ O), poured into a separatory funnel, shaken with dichloromethane (CH ₂ Cl ₂ ) and equilibrated (the remaining 70% EtOH concentrate was stored).

(5) The dichloromethane layer (lower layer) was collected, treated with sodium sulfate, filtered and concentrated, and ethyl acetate (EtOAc) was added to the water layer and then equilibrated.

(6) The ethyl acetate layer (upper layer) was collected, treated with sodium sulfate, filtered and concentrated, and butanol (BuOH) was added to the water layer and then shaken and equilibrated.

⑦ Butanol layer (upper layer) and water layer (lower layer) were separately collected and treated with sodium sulfate, followed by filtration and concentration to obtain 48.71 g and 194.4 g (FIG. 1).

(IF), butanol fraction (BF) and water fraction (HF) except ethyl acetate and water layer in the fractionation of ethyl acetate were obtained as shown in FIG. 1 Respectively.

As a positive control substance, a composition obtained by concentrating GGEx18, which is a conventional invention substance, with hot water extraction (GG18) or ethanol (EGG18) (1.5 L each for 6 times and 2 hours) was used in the experiment.

Experimental Example  1. Animal test design to identify anti-obesity and anti-diabetic effects

Experimental Example 1 -One. Preparation of experimental animals

As a public animal, 8-week-old C57BL / 6N male mice supplied from Goyonggi province, South Korea were used. Eight groups per group were randomly divided into groups according to body weight range and used for the experiment. The incubation environment was 21 ± 2 ℃ for temperature, 55 ± 5% for humidity, 15 ~ 17 times / hour for ventilation, 150 ~ 300 lux for illumination and 12 hours for light (light: 06:00, light: 18:00) And maintained in a SPF (specific pathogen free) state during the experiment. Solid feed (Harlan, USA) and water were fed free and watered.

Experimental Example 1 -2. Experimental group  And administration method

Eight male males were randomly assigned to receive a 50 mg / kg dose for each fraction and a 250 mg / kg dose for 9 weeks. To check for obesity and diabetes, the C57BL / 6N group, a normal control group, was fed with low fat, and no drug was given to the control group of the high fat group. (Table 1).

Group Treatment Number Sex Normal Low fat 8 male High fat + extract or fraction (mg / kg BW) Control 0 8 male CF 50 8 male EF 50 8 male IF 50 8 male BF 50 8 male HF 50 8 male EGGEx18 250 8 male GGEx18 250 8 male

Experimental Example  2. Effect on weight

Experimental Example  2-1. Effect on weight gain

The body weight was measured twice a week during the 9 - week experimental period to confirm the effect on body weight gain.

As shown in FIG. 2 and FIG. 3, in the high fat feed control group, the body weight was increased by about 40% as compared with the general feed control group. On the contrary, in the group treated with 250 mg / kg of hydrothermal extract of Kang Shin Jang, the weight gain was about -23.18%, -37.14% in the BF group, -27.32% in the CF group, 30.24%, -26.74% in the IF group, -17.52% in the HF group, and it was confirmed that the composition had an effect of suppressing weight gain. Particularly, in the case of the fraction-administered group, it was confirmed that it could act as a preventive and therapeutic agent for obesity because it had a better effect of inhibiting weight gain, even though it was 1/5 of the capacity of the hot-water extract of the present invention.

Experimental Example  2-2. In food efficiency  Effect

To determine whether the test substances were related to appetite, body weight was measured twice a week, feed intake was measured once a week for 9 weeks, and the dietary efficiency was calculated.

Dietary efficiency was calculated by dividing the final weight gain by the total feed intake by substituting into the formula below.

Feed efficiency ratio (FER%) = weight gain (g) / feed intake (g) × 100

As indicated by the formula of diet efficiency, the low body weight gain despite the high intake of diets may be considered to have an obesity modulating effect. Therefore, the food efficiency can be used as a measure of obesity, the smaller the value of the food efficiency can be said to have an obesity control effect.

As shown in FIG. 4, about 50.68% of the dietary efficiency was higher in the high-fat feed control group than in the general feed control group, and it was confirmed that the high fat diets resulted in obesity. In contrast, in the GG18 group treated with 250 mg / kg of hydrothermal extract of the former inventor, Kang Ji-hwan Kang Ji-hwan, the dietary efficiency was 22.76% lower than that of the high fat feed control group. In the fraction BF group, 34.22%, CF group 26.24% 25.08% in group, 23.08% in IF group and 14.18% in HF group. Particularly, in the case of the fraction-administered group, it was confirmed that even though it is 1/5 of the capacity of the hot-water extract of the present invention, the fraction has a lower dietary efficiency than that of the hot-water extract of the present invention.

Experimental Example  3. Measuring Fat Tissue Weight Change

The mice of Experimental Example 2 were slaughtered and then weighed to measure the peripheral white adipose tissue (EAT), the white adipose tissue around the abdominal wall (RAT), the inguinal subcutaneous adipose tissue (IAT) and the brown adipose tissue (BAT) .

As shown in FIG. 5, the weight of four adipose tissues was significantly increased in the high-fat feed control group compared with the general feed control group, indicating that obesity occurred. On the contrary, EAT, RAT, and BMI were significantly higher in GG18 group and BF group, CF group, EF group, IF group, and HF group treated with 250 mg / kg of hydrothermal extract of the former invention, IAT and BAT weight were all decreased. Particularly, in the case of the fraction-administered group, the weight of the fat tissue was lower than that of the hot-water extract of the present invention, which was 1/5 of the conventional hot-water extract.

Experimental Example  4. Histology of Adipose Tissue speculum

The fatty tissue isolated from the mouse of Experimental Example 2 was fixed in 10% phosphate-buffered formalin for one day. The formalin was washed with water for 12 hours or more, and then washed with 60% ethanol for 1 hour and 70% 1 hour, 1 hour in 80% ethanol, 1 hour in 90% ethanol, 1 hour in 95% ethanol, and 1 hour in 100% ethanol. Xylen was permeabilized three times per hour for 1 hour and then infiltrated 2 times for 1 hour to paraffin. After embedding, the slices were sectioned to a thickness of about 3 쨉 m. Fat tissues were laid on the slides and dried, followed by hematoxylin-eosin (H & E) staining. The slide was drained and the mounting solution (Sigma, MO, USA) was dropped to allow permanent preservation. Photographs of the tissues and the number and size of adipocytes were analyzed using an image analysis system (Image Pro-Plus, MD, USA).

As shown in FIGS. 6 and 7, it can be seen that the size of the fat cells of the high-fat diet control group was significantly increased (137%) as compared with that of the general diet control group. In contrast, in the fraction-treated group, the fat cells were statistically significantly smaller than those in the high-fat feed control group in the BF group, -37%, -30%, and -41% I can confirm that it is gone. In particular, it was found that the fraction-treated group had a better anti-obesity effect even at a low dose of 1/5 compared to the hot-water extract administered group (-25%).

Experimental Example  5. Histology of Liver Tissue speculum

The liver tissue was treated in the same manner as in Experimental Example 3 to observe the fat content in liver tissue. As shown in FIG. 8 and FIG. 9, in the case of the high fat feed control group, the fat content in the liver tissue was increased by about 600% as compared with that in the general feed control group. On the other hand, in the fraction-treated group, -86% in the BF group, -86% in the CF group, 86% in the EF group, -86% in the IF group, and -86% It can be confirmed that it is effectively suppressed.

Experimental Example  6. Impact on liver function

For the purpose of biochemical analysis of mouse blood, mice were fasted for 12 hours and then anesthetized with diethyl ether. Then, 1 ml of blood was collected from the abdominal vein and centrifuged in a high speed centrifuge (Micro 12, (AST) and alanine aminotransferase (ALT) were measured using a blood biochemical analyzer (Selectra 2, Vitalab, Netherlands) after centrifugation at 5,000 rpm for 10 minutes. Were measured.

AST and ALT are indicators of hepatic abnormality, and the increase of the enzyme indicates that there is liver damage.

As shown in FIG. 10, there was no statistically significant increase or decrease in AST in all groups. The ALT increased in the high-fat diet control group compared to the general diet control group, whereas the total extract and fraction administration group , Respectively. Through this, it was confirmed that the whole extracts and fractions of Kyungshingangjihwan showed the effect of treating obesity and diabetes without damaging liver tissue.

Experimental Example  7. Diabetes improvement effect

Serum glucose and insulin blood levels were measured in the same manner as in Experimental Example 6.

As shown in Figure 11 it can be seen that the fasting blood glucose in the high fat feed control diabetes induced more than 190 mg / dL. In the group administered with 250 mg / kg of the whole extract of Kyungshingang Jihwan, there was no statistically significant change compared to the control group of the high fat diet, but in the group administered with the fraction group, BF, CF, EF, IF at 50 mg / kg, It was found to be reduced by blood glucose in the group fed the diet.

In addition, as shown in FIG. 12, the fasting insulin level was significantly increased in the high fat feed control group compared to the normal feed control group, and it was confirmed that type 2 diabetes due to insulin resistance occurred. On the other hand, the group treated with the fraction-treated group BF, CF, EF and IF at 50 mg / kg showed a statistically significant decrease in insulin levels in the group fed the normal feed.

Through the above results, it was confirmed that the fractions of the gangsinjigi can effectively act in the prevention and treatment of diabetes.

Experimental Example  8. Indicator Component Analysis

For each fraction, the ratio of ependrine to sennoside A was analyzed (Table 2).

Ephedrine results Sennoside A result HF: 0.26% HF: 0.07% BF: 4.06% BF: 0.16% IF: 1.0% IF: 0.14% EF: 0.71% EF: 3.33% CF: 0.91% CF: 0.1% EGG18: 1.30% EGG18: 0.48% GG18: 0.81% GG18: 0.02%

Hereinafter will be described a pharmaceutical composition comprising the composition of the present invention and a preparation example of a health functional food, the present invention is not intended to limit this but only to be described in detail.

Formulation example  One. Sanje  Produce

Fraction of Kangshin Kang Ji Hwan 2 g

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

100 mg of the fractions of Kangshin Kang Ji Hwan

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3. Preparation of capsules

100 mg of the fractions of Kangshin Kang Ji Hwan

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

100 mg of the fractions of Kangshin Kang Ji Hwan

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ .2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

Formulation example  5. Liquid  Produce

100 mg of the fractions of Kangshin Kang Ji Hwan

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example  6. Manufacture of health food

1 g of fraction of Kangshin Kang Ji Hwan

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7. Health drink  Produce

1 g of fraction of Kangshin Kang Ji Hwan

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (19)

delete delete delete delete delete delete delete 50% (v / v%) of Kyungshingang Jihwan 18 (GGEx18) prepared by mixing ephedra, bales and rhubarb in a weight ratio of 10% to 50%: 20% to 50%: 20% to 50% based on the weight of the raw herbal medicine ) To 80% (v / v%) extracted and concentrated with ethanol, and fractions obtained by dividing it in the order of dichloromethane, ethyl acetate, butanol and water; pharmaceutical composition for the prevention and treatment of diabetes comprising as an active ingredient Composition. delete The composition of claim 8, wherein the fraction is a dichloromethane fraction. 9. The composition of claim 8, wherein the fraction is an ethyl acetate fraction. The composition of claim 8 wherein the fraction is a butanol fraction. The composition of claim 8, wherein the fraction is an intermediate fraction excluding the ethyl acetate layer and the water layer of the fraction fractionated with ethyl acetate. The composition of claim 8, wherein the diabetes is insulin independent diabetes due to insulin resistance. 50% (v / v%) of Kyungshingang Jihwan 18 (GGEx18) prepared by mixing ephedra, bales and rhubarb in a weight ratio of 10% to 50%: 20% to 50%: 20% to 50% based on the weight of the raw herbal medicine ) To 80% (v / v%) extracted with ethanol and concentrated, dichloromethane, ethyl acetate, butanol and fractions obtained by separating the system in the order of water; preventive and improved health of diabetes comprising as an active ingredient Nutraceutical. The composition of claim 10, wherein the dichloromethane fraction comprises 0.5 to 3 wt% of ephedrine and 0.05 to 0.3 wt% of senosides A as indicators. The composition of claim 11, wherein the ethyl acetate fraction comprises 0.3 to 1.5 wt% of ephedrine and 1.5 to 6 wt% of sennoside A as indicators. The composition of claim 12, wherein the butanol fraction comprises 2 to 8% by weight of ephedrine and 0.05 to 0.3% by weight of senosides A as indicators. The composition of claim 13, wherein the intermediate fractions other than the ethyl acetate layer and the water layer contain 0.5 to 2 wt% of ephedrine and 0.05 to 0.3 wt% of sennoside A as indicators.

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