KR100473529B1 - Composition comprising an extract of sungisan crude drug complex as an effective ingredient for preventing and treating diabetes - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of diabetic herbal medicine complexes, that is, extracts of hubac, rhubarb and fat, as an active ingredient and excellent antidiabetic activity, the composition of the present invention suppresses weight gain and effectively prevents blood sugar. It does not show hepatotoxicity while lowering, and thus can be used as a medicament and dietary supplement useful for the prevention and treatment of diabetes mellitus and its various complications.

Description

COMPOSITION COMPRISING AN EXTRACT OF SUNGISAN CRUDE DRUG COMPLEX AS AN EFFECTIVE INGREDIENT FOR PREVENTING AND TREATING DIABETES}

The present invention relates to a composition for preventing and treating diabetes mellitus, which contains pure acidic herbal medicines, that is, extracts of hubac, rhubarb and fruit, as active ingredients, and has excellent antidiabetic activity.

Recently, the food culture of Korean people has been influenced by Western civilization, such as the United States, so that the rapid transition from low-fat diet to high-fat diet is steadily increasing in children's obesity population. In the case of excessive food intake, the amount of exercise is significantly reduced, the situation is aggravating the problem of obesity.

Syndrome X refers to a condition in which hypertension, hyperlipidemia, obesity (particularly the abdominal region) and insulin resistance are simultaneously present, not a specific disease but a combination of a series of related risk factors that often develop simultaneously. (Susan AE et al ., Diabetes Care 21: 1637-1643, 1998). Therefore, people with syndrome X are at high risk of developing diabetes or diseases of the circulatory system. As the obesity population increases, the incidence rate of adult diseases characterized by X syndrome is steadily increasing, so appropriate measures are required.

Type 2 diabetes (Adult Diabetes, Type 2 diabetes) accounts for over 90% of diabetic patients and is a disease that is more likely to develop after 40 years of age, especially in obese people, characterized by insulin resistance. People with insulin resistance develop hyperinsulinemia because they regulate high blood sugar to normal blood sugar by releasing too much insulin. Early physiological adaptation to insulin resistance is hyperinsulinemia caused by excessive proliferation of β cells, but it does not release enough insulin to overcome the resistance of peripheral tissues to insulin, resulting in a repeated cycle of hyperglycemia and hyperinsulinemia. Eventually resulting in necrosis of β cells, insulin deficiency, severe hyperglycemia, weight loss, etc. (Defronzo RA, Diabetes Rev. 5: 177-269. 1997; Polonsky KS et al ., New Engl. J. Med . 334: 777-783, 1996; Reaven GM, Physiol. Rev. 75: 473-486, 1995).

For the treatment of type 2 diabetes, clinicians have mainly used sulfonylurea drugs that promote insulin secretion for the purpose of lowering blood sugar (Lebovitz HE, Diabetes Rev. 7: 139 ~ 153, 1999). Since insulin levels of patients with insulin resistance are not inferior to those of normal subjects, administration of drugs that promote insulin secretion may lower blood sugar at the beginning of treatment, but it is a long-term benefit. . Indeed, when sulfonylurea-based drugs are administered for a long time to lower blood sugar of type 2 diabetic patients, they gain weight and do not show any effects over time. Therefore, insulin injections are often required. have.

For the purpose of supplementing these shortcomings of sulfonyl urea drugs, three drugs of troglitazone (troglitazone, rosiglitazone, pioglitazone), which are called insulin resistance improving agents or insulin sensitizers, have recently been developed and attracted attention from Sein. Day C., Diabetes Med . 16: 1-14, 1999). Unlike the sulfonyl urea drugs, these drugs are known to be effective in improving insulin resistance by helping insulin work in muscle and adipose tissue without affecting insulin secretion. In terms of efficacy, there are some improvements compared to existing drugs, but the first triglitazone released in 1997 as a drug of this type is severely toxic to the liver and is currently forbidden to be sold. Rosiglitazone and pioglitazone ( pioglitazone is a drug that has been released recently (May and July 1999, respectively) and has not yet secured long-term safety. Other oral hypoglycemic agents include biguanide metformin (Bailey CJ et al ., New Engl. J. Med. 334: 574 ~ 579. 1996) and acarbose, a drug that delays glucose absorption in the small intestine (acarbose) (Lebovitz HE, Diabetes Rev. 6: 132 ~ 145, 1998) are used, but in the former, there is a risk of side effects such as lactic acidosis, and in the latter, diabetes is not well controlled after meals. There are limitations for use in patients (Bailey CJ, Biochem. Pharmacol . 58: 1511-1520, 1999; DeFronzo RA, Ann Intern. Med . 131: 281-303, 1999).

On the other hand, the research direction of new drug development using natural products in Korea has been focused on clarifying their chemical effects and identifying chemical structures by separating active substances from single herbal or folk medicines, but the results are very small. Many prescriptions described in traditional medicines are effective for various diseases and are still used by many domestic and foreign Chinese doctors and the general public. Identifying the medicinal effects of unilateral herbal medicines and separating the active ingredients from them into drugs (for example, several antibiotics, lovastatin, which is used for treating hyperlipidemia, and taxol, which is an anticancer agent), are also meaningful, but they are actually a single herbal medicine in Korea. There is no case of developing diabetes treatment from folk medicine). However, if it is possible to evaluate the efficacy of the prescriptions made of several Chinese medicines, reveal the mechanisms, and finally derive new prescriptions from them, this method would be more competitive in competition with leading foreign pharmaceutical companies.

Therefore, the present inventors conducted a intensive study to develop a preventive and therapeutic agent for diabetes using natural products, and extracts of the herbal extracts of narcotic herbs, rhubarb, and larvae in ob / ob mice of type 2 diabetes with obesity and hyperinsulinemia The present invention has been completed by revealing that it can be usefully used for the prevention and treatment of diabetes due to the suppression of weight gain and excellent blood glucose lowering effect and no liver toxicity.

An object of the present invention is to provide a pharmaceutical composition and dietary supplement showing the effect of preventing and treating diabetes and its complications using natural products.

In accordance with the above object, the present invention provides a composition for the prevention and treatment of diabetes mellitus and thereby complications, including pure acidic herbal compound combinations, that is, extracts of hunch, rhubarb and fruit.

Hereinafter, the present invention will be described in detail.

Type 2 diabetes is a steadily increasing disease due to the development of modern society and the westernization of diet. It is closely related to insulin resistance and causes a number of fatal complications due to persistent hyperglycemia. Therefore, improvement of insulin resistance and regulation of hyperglycemia to normal blood glucose in type 2 diabetes are the primary treatment goals.

To this end, the present invention provides a composition for the prevention and treatment of diabetes mellitus containing a pure herbal acid (생) herbal combinations, that is, extracts of hubak, rhubarb and fruit as an active ingredient.

The composition for preventing and treating diabetes of the present invention comprises 0.5 to 50% by weight of the extract of pure acid herbal medicine combination based on the total weight of the composition.

Each herbal medicine of the present invention includes all the herbal medicines derived from the same plant, and the composition ratio of the individual compositions is not limited thereto, but preferably, the weight composition ratio of thick, rhubarb, and fat fruit is about 1-10: 1 to 1. 8: 1, more preferably about 3-7: 2-3: 1, most preferably 2.5: 1: 1.

After extracting all the herbal extracts of the present invention, the seeds are crushed and the roots or roots are chopped and then about 1 to 15 times the volume of the herbal medicine, preferably about 5 to 10 times the water, lower alcohol or A mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably 1: 1 to 1: 3, at about 20 to 100 ° C., preferably at about 70 to 100 ° C. for about 1 hour to 2 days, Preferably it can be obtained by extraction methods such as ultrasonic extraction, reflux extraction in about 2 hours to 1 day.

The present invention provides a composition for the prevention and treatment of diabetes mellitus and its complications, including water extracts obtained in the extraction process, lower alcohol extracts and soluble extracts according to a mixed solvent thereof.

Machilus thumbergii is a dried bark of a magnolia and a plant of a walnut tree, or a bark of thick branches. It is plank-shaped or semi-tubular, and the surface is grayish white or grayish brown. write. Muscle stiffness, cramps, bloating of the abdomen, infiltration, antibacterial, soothing, healthy. However, no antidiabetic activity has been reported.

Rhubarb (大黃, Rheum undulatum Linne ) is a perennial plant, removed from the roots and burdock-shaped roots of 6 to 7 years old, and then chopped and dried as it is. In oriental medicine, it has been used as an anti-inflammatory laxative since BC and has been used in various prescriptions. Main components are anthraquinone derivatives, glycosides and tannins thereof. If you eat a small amount of health (健 위 作用) indicates a large amount of intake as a laxative (緩 下 劑) is good for constant constipation or indigestion, and folk medicine is also used for burns. When medicinal is used to take a month to take raw and strong, and when you use ripe, the effect is gentle and char-dried is strong hemostasis. However, no antidiabetic activity of rhubarb has been reported. In general, the golden gate rhubarb contains a lot of sennoside-type components showing the lower action, so in order to exclude the non-specific blood sugar lowering action by the lower action, the present invention uses the lower rhubarb action than the golden moon rhubarb do.

Poncirus trifoliata Rofinesque ( Poncirus trifoliata Rofinesque ) is an immature fruit of citrus fruits such as tangerines, stirrups and summer citrus fruits, cut in half or dried, and has a thick fruity and bitter taste. Dried ripe fruit is called "perception" and is equivalent to fruit in terms of efficacy. Its main ingredient contains isosakuranin, naringin, citriforlioside, poncirin, etc. Obesity swelling, expectoration, news, small intestine, sexual harassment, and ulcerative swelling脹滿), gastrointestinal lethargy, indigestion, allergic constitution, gastric gas removal, uterine sewage, visceral laxity. However, no antidiabetic activity has been reported.

In addition, Donggibogam Sogalmun is a small and medium, characterized by a thin, spontaneous sweat and a good stool without the stool coming out of it. Although it has been used in the description, the anti-diabetic activity of the extract of pure herbal medicine herbal extracts composed of these herbs will also be identified for the first time in the present invention.

When the pure acid extract of the present invention is administered to ob / ob mice of type 2 diabetes model, dietary and drinking water intake is reduced and dose-dependent weight gain is suppressed. Compared to the diabetic control group, the pure acid group showed a dose-dependent hypoglycemic effect while maintaining a low blood sugar level from the first week of the sample administration, and the urine glucose excretion was also reduced. In addition, pure Gi acid not only controls blood glucose steadily throughout the administration period by reducing blood glycated hemoglobin level by more than 1.0%, but also lowers insulin and C-peptide levels in a dose-dependent manner, thereby improving hyperinsulinemia and insulin resistance in ob / ob mice.

Pure acid also has the effect of significantly reducing the levels of total cholesterol, HDL, LDL, free fatty acids, triglycerides, etc. in ob / ob mice to improve insulin resistance by regulating lipid metabolism.

As such, the pure acid extract of the present invention exhibits a weight-inhibiting effect and a pronounced hypoglycemic activity in ob / ob mice of type 2 diabetes animal model. The hypoglycemic activity of narcotic acid improved the hyperinsulinemia, insulin resistance and blood lipid levels that are characteristic of ob mice. In addition, since the enzyme activity of ALT and AST, which are indicators of liver toxicity, belongs to the normal range in the pure acid-administered group, the pure acid of the present invention confirms that it is a very safe substance that does not exhibit liver toxicity.

As confirmed above, a composition comprising pure acid extract, which has been proven to have very high human safety due to its excellent anti-diabetic activity and no hepatotoxicity, is useful for the prevention and treatment of diabetes mellitus and its various complications. Can be used. Here, but not limited to, complications due to diabetes may be a disease due to the increase in blood concentration of triglycerides, cholesterol, high density lipids.

The composition of the present invention containing pure acid extract as an active ingredient can be administered orally or parenterally during clinical administration and can be used in the form of general drug replacement therapy.

That is, the composition may be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used, may be used. It is prepared by. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, and the like. (Sucrose) or lactose (Lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium, stearate and talc are also used. Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized replacement treatments, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The compositions of the present invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

Typical daily dosages of pure acid extracts of the present invention range from 100 to 1,000 mg / kg body weight, preferably from 200 to 500 mg / kg body weight, and may be administered once or in divided doses. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex, weight of the patient and the severity of the patient, and thus the dosage should in any way be within the scope of the invention. It is not intended to limit.

The composition containing the pure acid extract of the present invention may be used in various forms, such as drugs, foods and beverages for the prevention and treatment of diabetes mellitus and its complications. Examples of the food to which the pure acid extract of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health supplements.

Pure acid extract of the present invention itself has little toxicity and side effects, so can be used with confidence even for long-term administration for the purpose of prevention.

The amount of pure acid extract contained in the food or beverage may be added at 0.5 to 50% by weight, preferably 10 to 20% by weight of the total food weight, and the food health beverage composition may be 0.5 to 10 g based on 100 ml, Preferably it can be added in the ratio of 1-5 g.

The health beverage composition of the present invention contains the above ingredients in the ratios indicated and may contain various flavors or natural carbohydrates and the like as additional ingredients, as in conventional beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (taumartin), stevia extracts (e.g., Rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.) are advantageously used. Can be. The proportion of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of about 0-20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Preparation of Pure Acid Extract

To prepare extracts of Sugisan (SGis) herbal medicines, 10 times the amount of water was added to 10.0 g of rhubarb, 8.0 g of rhubarb, and 4.0 g of fermented soybeans, which were purchased from the herbal medicine distributor Yerang Building Co., Ltd. It was heated to reflux for 2 hours at a temperature of ℃. The extract obtained therefrom was lyophilized using a lyophilizer (FN-5N, EYELA, Japan) to use 100 g of a pure acid herb composite powder obtained by drying as a sample.

Example 2 Analysis of Physiological Changes Caused by Administration of Pure Acid

(2-1) Laboratory Animals and Diet

In order to investigate the antidiabetic activity of the extract of the pure herbal extract, the male C57BL / 6J ob / ob mice (The Jackson Laboratory) were used as the diabetic control group and the experimental group, and the C57BL / 6J black mouse (the central experimental animal) was used as the normal control group (lean). The following experiment was carried out using the control). ob / ob mice are genetic hyperglycemic obesity mutants that appear when the recessive gene ob on the sixth chromosome becomes homozygous (ob / ob). Obesity is seen at three weeks of age, after which the weight gain is noticeably increased to 80 g, about three times the normal mouse weight. Ob / ob mice develop insulin resistance with increasing body fat and continue to have hyperglycemia and hyperinsulinemia, but blood sugar levels and insulin levels usually drop to normal levels in the later stages of obesity. In the early stages of obesity with hyperglycemia and hyperinsulinemia, pancreatic hypertrophy due to the hyperplasia of pancreatic β cells is remarkable.

After 6 weeks of ob / ob mice were acclimated to the laboratory environment for 1 week, the pure acid extract prepared in Example 1 was administered at 200 or 400 mg / kg dose for 9 weeks. The experimental animals were bred in animal kennels maintained at a humidity of 50% and a temperature of 24 ~ 26 ℃, and water was supplied for free intake.

Six rats were divided into four groups based on blood glucose and body weight before administration of pure gisan extract as follows: Normal control rats and ob / ob mouse diabetic rats were fed with powdered mice only (Samyang). The administration group was supplied with a mixture of pure acid extract to feed to 200 mg or 400 mg per kg body weight of the mouse in two doses. Sample dose was determined by examining the amount of food for 3 days before starting the administration, and the sample volume was changed every two weeks according to the weight and diet.

All experimental results were expressed as the mean value ± S.E. and statistical processing was performed by the Student t-test to determine the significance based on p <0.05.

(2-2) Diet and Drinking Water Intake Changes

Table 1 shows the changes in dietary and drinking water intake after administration of pure Gisan extract. In the diet and drinking water intake, the diabetic control group was significantly increased compared to the normal control group, whereas the pure acid group was significantly decreased in the 200 mg and 400 mg groups compared to the diabetic control group. In particular, drinking water intake showed a dose-dependent decrease of 45% and 52% in the pure acid group compared to the diabetic control group, respectively. This indicates that the pure acid extract has an activity to improve the symptoms of diabetes mellitus, Dagal.

Normal control Diabetes control Pure acid treatment group  200 mg 400 mg Dietary Intake (g / mouse / day) 3.0 ± 0.3 4.7 ± 0.3 ⁺⁺ 3.5 ± 0.2 ^* 3.7 ± 0.4 ^* Drinking water intake (ml / mouse / day) 4.7 ± 0.1 7.9 ± 0.4 ⁺⁺ 4.4 ± 0.4 ^** 3.8 ± 0.8 ^**

(2-3) weight change

1 is a graph showing the weight change due to the administration of pure acid extract. As shown in FIG. 1, the diabetic control group gained 21.6 g of body weight for 9 weeks, while the body weight of 200 mg group was 12.6 g, 9.2 g group of 400 mg group, and 5.3 g of normal group. As the percentage of weight gain in each group, the diabetic control group increased 59% compared to the weight before sample administration, and the normal control group increased 23%, and the 200 mg group increased 38% and the 400 mg group increased 25%. Dependently, it showed the activity of inhibiting weight gain, and especially, the 400 mg administration group showed weight gain inhibition activity of the normal control level. These results suggest that pure acid extract inhibits weight gain by inhibiting the polyphagia symptoms that are characteristic of ob / ob mice (Table 1).

(2-4) change in fat weight

Table 2 shows the results of comparing fat weight to body weight of each group after the administration of pure acid extract. Compared with the normal control group, the fat weight of the diabetic control group was significantly increased, and the fat weight of the pure acid group was not significantly different.

Normal control Diabetes control Pure acid treatment group   200 mg  400 mg Body weight (g) 27.5 ± 1.9 57.1 ± 2.4 ⁺⁺ 49.5 ± 1.5 ^** 44.9 ± 0.8 ^** Fat content (g) 0.43 ± 0.06 3.26 ± 0.29 ⁺⁺ 3.38 ± 0.25 3.25 ± 0.13 Fat hypertrophy index (%) 1.59 ± 0.28 5.72 ± 0.44 ⁺⁺ 6.82 ± 0.40 ^** 7.24 ± 0.22 ^**

(2-5) height change

Table 3 is a result of comparing the weight of the kidney to the weight of each group after the administration of pure acid extract. The weight of the kidney was significantly increased in the diabetic control group compared to the normal control group, but the dose was not dose-dependent but significantly decreased in the diabetic control group compared to the diabetic control group. Values did not show any difference between the diabetic control group and the pure acid group.

Normal control Diabetes control Pure acid treatment group 200 mg 400 mg Body weight (g) 27.5 ± 1.9 57.1 ± 2.4 ⁺⁺ 49.5 ± 1.5 ^** 44.9 ± 0.8 ^** Kidney Weight (mg) 154 ± 13 196 ± 22 ⁺ 158 ± 28 ^** 154 ± 4 ^** Renal hypertrophy index (%)  0.56 ± 0.02 0.34 ± 0.03 ⁺⁺ 0.32 ± 0.05 0.34 ± 0.01

Example 3 Analysis of Biochemical Changes Caused by Administration of Radical Acid Extract

(3-1) Blood Body Extraction and Treatment

Blood collection for blood marker analysis was performed at 3 pm every week. Plasma obtained by collecting blood through the orbital vein without anesthesia and then centrifuging at 5,000 rpm for 10 minutes was used in the following experiment.

(3-2) Hypoglycemic Effect of Pure Radix Extract

In order to measure the hypoglycemic effect of the extract of pure acid, the plasma obtained in the intraorbital vein weekly as shown in Example (2-1) using glucose-oxidation method (glucose-oxidation method, Trinder, Sigma) Concentrations were measured (DS Young, Editor: Effects of drugs on clinical laboratory tests, 3rd Ed. AACC Press , Wsshing, 1990; Bergmeyer HU et al ., Methods of Enzymatic analysis 2nd Ed .: 1205-1212, 1974).

As a result, as shown in Figure 2, the diabetic control group maintained a high blood sugar of 250,300 mg / 내 throughout the experimental period and showed a significant difference compared to the normal control group. On the other hand, naive acid treated group showed a hypoglycemic activity compared to the diabetic control group from 1 week of administration, and maintained blood glucose similar to that of the normal control group throughout the administration period and showed dose-dependent hypoglycemic activity. Considering the pre-administration period, the 200 mg group showed a significant difference compared to the diabetic control group except the 4, 5 and 6 weeks of administration, and the 400 mg group was significantly different from the diabetic control group except the 4 weeks. Difference was shown. At 15 weeks of age (the last week of administration), the blood glucose level of the diabetic control group was 247 mg / dL, whereas the 200 mg and 400 mg pure basal groups were 129 and 117 mg / dL, respectively, with dose-dependent doses of 48% and 53%, respectively. Phosphorus blood sugar drop rate was shown, indicating the activity of dropping the blood sugar level to a level lower than 177 mg / dl of the normal control group used in the experiment.

(3-3) Serum Glycosylated Hemoglobin Concentration by Pure Acid Extract

In order to investigate the change in blood glycated hemoglobin concentration by pure acid extract, the blood glycated hemoglobin level was obtained using the cation-exchange method (Glycated hemoglobin HbA1 kit, Sigma) in whole blood collected from mice at the last week of sample administration. Was measured (Boil G. et al ., Diabetes 29: 272, 1980).

Glycated hemoglobin is a measure of long-term changes in blood glucose, and as shown in FIG. 3, the diabetic control group had an average glycated hemoglobin level of 4.65%, higher than that of the normal control group, 3.58%, whereas the pure group was 3.45% and 3.41%, respectively. There was a dose-dependent significant difference compared to the diabetic control group and the glycated hemoglobin level was lowered to the normal control level. These results are consistent with the results of hypoglycemic activity of the pure acid extract shown in Figure 2, which is a 1.20% reduction in glycated hemoglobin level compared to the diabetic control. Considering that it is common to determine that the hypoglycemic activity of the sample is clear when administration of the glycated hemoglobin level is more than 1% after 9 weeks of administration, it can be seen that the pure acid extract effectively regulates blood glucose.

(3-4) Changes in Blood Insulin Concentration by Pure Acid Extract

Plasma obtained from the orbital vein at the last week of sample administration was measured using an insulin radioimmunoassay kit (Coat-A-Count Insulin, DPC).

As shown in FIG. 4, the insulin level after the end of the administration was 1.46, 75.58, 25.68 and 14.36 μIU / ml in the order of the normal control group, the diabetic control group, and the 200 mg and 400 mg pure acid groups. The diabetic control showed hyperinsulinemia, which is insulin resistant with increasing blood sugar and body fat, and continues to show hyperglycemia and hyperinsulinemia, consistent with the previously reported characteristics of ob mice, 200 mg and 400 In the mg-naive acid group, the insulin level was lowered by 64% and 81%, respectively, compared to the diabetic control group, and it was determined that narcotic extract regulates blood glucose by improving hyperinsulinemia and insulin resistance, which are characteristic of ob mice.

(3-5) Changes of C-peptide Concentration in Blood by Extracts

Plasma obtained from the orbital vein at the last week of sample administration was measured for blood C-peptide concentration using a double antibody C-Peptide kit (EURO DPC).

As shown in FIG. 5, the C-peptide level after the end of the administration was 0.09, 0.70, 0.27 and 0.13 ng / ml in the order of the normal control, diabetic control, 200 mg and 400 mg pure acid administration group. C-peptide levels in blood can be used to accurately determine insulin production and secretion. C-peptide levels, like insulin concentrations, showed a dose-dependent decrease in pure acid-administered groups.

(3-6) Changes in Urinary Glucose Excretion by Pure Acid Extract

In the last week of sample administration, urine glucose levels were measured from glucose from each group of animals using the glucose-oxidation method (Trinder, Sigma).

As shown in Table 4, the urine glucose concentration was 717.6 mg / dl in the diabetic control group, a 35-fold increase compared to the normal control group (19.6 mg / dl), while the pure acid group was 16.3 and 19.1 mg / dl, compared with the normal control group. Similar values were observed with changes in blood glucose levels.

Normal control Diabetes control Pure acid treatment group 200 mg 400 mg Urinary glucose (mg / dl) 19.6 ± 3.2 717.6 ± 144.3 ⁺⁺ 16.3 ± 2.0 ^** 19.1 ± 2.1 ^**

(3-7) Changes in Serum Lipid Concentration by Pure Acid Extract

Lipid metabolism disorders are closely related to type 2 diabetes, especially high free fatty acids and triglycerides in blood are closely related to insulin resistance in skeletal muscle and adipose tissue. Therefore, the change of blood lipid concentration was measured to investigate the effect of pure acid extract on lipid metabolism and the resulting insulin resistance improvement.

Total cholesterol levels of cholesterol in the plasma obtained from the orbital vein was -E kit using (Cholesterol E-Kit, pharmaceutical gel) measurements every 3 weeks, and serum HDL cholesterol level HDL bikbeok isopropyl ^TM spin kit (BIG BUCK HDL ISOSPIN ^TM kit, Sigma) (Barr DP et al ., Am J Med 11: 480, 1951). In addition, the blood triglyceride concentration was measured using a GPO-Trinder kit (Sigma), and the concentration of nonesterified fatty acid (NEFA) in the blood was measured using an acyl-CoA oxidation method ( It was measured using the acyl-CoA oxidase method, NEFAZYME-S, Eiken (PNM Demacker. et al ., J Clin. Chem . 28 (8): 1765, 1982).

As shown in Table 5, the total cholesterol, LDL, and free fatty acid levels of the diabetic control group were significantly increased compared to the normal control group, whereas the pure acid group received total cholesterol levels at the 200 mg and 400 mg doses, respectively. The levels of 32% and 30% and free fatty acids were reduced by 26% and 16%, respectively. In addition, triglyceride levels were significantly lower than the diabetic control group by 27% in the 200 mg administration group, and improved HDL and LDL levels. Compared with the pure acid group, the 200 mg group showed more effective blood lipid level improving activity than the 400 mg group.

Normal control Diabetes control Pure acid treatment group   200 mg 400 mg Total Cholesterol (mg / dl) 142 ± 11 258 ± 20 ⁺⁺ 177 ± 11 ^** 183 ± 15 ^** HDL (% total cholesterol) 50.7 ± 3.7 47.7 ± 3.6 52.9 ± 4.0 ^* 49.2 ± 3.9 LDL (% of total cholesterol) 35.5 ± 4.5 44.2 ± 4.6 ⁺⁺ 38.5 ± 4.5 ^* 40.5 ± 4.4 Triglycerides (mg / dl) 97.0 ± 9.7 104.9 ± 13.0 76.1 ± 13.1 ^** 94.3 ± 12.4 Free fatty acid (μEq / ℓ) 1813 ± 95 1962 ± 166 ⁺ 1470 ± 164 ^** 1663 ± 133 ^*

(3-8) Changes in Total Blood Cholesterol Concentration by Pure Acid Extract

6 is a result of observing the effect of the pure acid extract on the total cholesterol level in the blood. As shown in Table 5, the pure acid extract showed a significantly lowering of total cholesterol level, and the pure acid group was shown to exhibit distinct activity from 3 weeks of administration, and the total cholesterol level was 150 mg / ㎗ throughout the administration period. In contrast, the diabetic control group had high total cholesterol levels of 250 mg / dL in the last week of the experiment.

(3-9) Blood ALT and AST Enzyme Activity

In order to confirm the hepatotoxicity of pure acid extract, the two representative enzymes, ALT (alanine transaminase) and AST (aspartate transaminase) enzyme activity, which are found in the blood during liver damage, were measured by UV ratio method from UV rate method, Asan Pharmaceutical).

As shown in Table 6, the diabetic control group showed increased activity in both enzymes compared to the normal control group, whereas the pure acid group showed low enzymatic activity at the normal control level at both doses, confirming that the pure acid extract was not hepatotoxic. Can be.

Normal control Diabetes control Pure acid treatment group 200 mg 400 mg AST (IU / ℓ) 4.4 ± 0.6 22.6 ± 6.2 ⁺⁺ 7.3 ± 1.3 ^** 6.3 ± 1.1 ^** ALT (IU / ℓ) 2.9 ± 0.9 32.8 ± 10.0 ⁺⁺ 6.2 ± 1.3 ^** 4.4 ± 0.8 ^**

Hereinafter, an example of the preparation of a pharmaceutical composition containing the pure organic acid herb extract of the present invention as an active ingredient will be described, but the composition of the present invention is not limited thereto.

Preparation Example 1 Preparation of Powder

According to the preparation method of powder in the Pharmacopoeia General Formulation, it is prepared in the following component content per packet.

Pure Acid Extract (Dry Powder) ㆍ ········ 300 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Talc 10 mg

Preparation Example 2 Preparation of Tablet

According to the preparation method of tablets in the pharmacopeia formulation, it is prepared in the following component content per tablet.

Pure Acid Extract (Dry Powder) ㆍ ········ 300 mg

Corn starch ㆍ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium stearate 2 mg

Preparation Example 3 Preparation of Capsule

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Pure Acid Extract (Dry Powder) ········ 300 mg

Corn starch ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium stearate 2 mg

Preparation Example 4 Preparation of Injection

According to the preparation method of injectables in the Pharmacopoeia General Formulation, it is prepared in the following component contents per ampule (2 ml).

Pure Acid Extract (Dry Powder) ㆍ ······· 300 mg

Sterile distilled water for injection ㆍ ·············

pH regulator ㆍ ··················

Preparation Example 5 Preparation of Liquid

According to the method for preparing a liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of the liquid formulation.

Pure acid extract (dry powder) ㆍ ·········· 1 g

Heterosaccharides ··············· 10 g

Mannitol ·················· 5 g

Purified water ㆍ ···················

As described above, the pure acid of the present invention suppresses blood glucose increase caused by diabetes and lowers the elevated levels of total cholesterol, HDL, and triglycerides, thereby exhibiting excellent antidiabetic activity and without liver toxicity, thereby preventing diabetes and its complications. It can be usefully used for prevention and treatment.

1 is a result of examining the effect of the pure acid extract of the present invention on the weight gain of ob / ob mice,

●: normal control ○: diabetes control

△: 200 mg pure acid administration group □: 400 mg pure acid administration group

2 is a result of examining the effect of the pure acid extract administration of the present invention on the plasma glucose concentration change of ob / ob mice,

●: normal control ○: diabetes control

△: 200 mg pure acid administration group □: 400 mg pure acid administration group

Figure 3 is a result of examining the effect of the pure acid extract administration of the present invention on the blood glycated hemoglobin concentration change in ob / ob mice,

○: glycated hemoglobin value-: mean value

4 is a result of examining the effect of the pure acid extract administration of the present invention on the blood insulin concentration change of ob / ob mice,

○: insulin value-: mean value

5 is a result of examining the effect of the pure acid extract administration of the present invention on the blood C-peptide concentration change of ob / ob mice,

○: C-peptide value-: mean value

Figure 6 is a result of examining the effect of the pure acid extract administration of the present invention on the change in total cholesterol concentration in blood of ob / ob mice.

Claims (5)

A pharmaceutical composition for preventing and treating diabetes mellitus and its complications, which is prepared by using the extract of the groin, rhubarb and fruit herb combinations as an active ingredient and a conventional pharmaceutically acceptable carrier. The method of claim 1, Complications due to diabetes is a pharmaceutical composition, characterized in that due to the increase in blood levels of triglycerides, cholesterol, high density lipids. The method of claim 1, Pharmaceutical composition, characterized in that the herbal extract is contained in 0.5 to 50% by weight relative to the total weight of the composition. The method of claim 1, Pharmaceutical composition, characterized in that the weight composition ratio of the thick foil, rhubarb and fruit room is 1-10: 1-1-10: 1-10. delete