KR20060018937A - Pharmaceutical composition comprising the extract of agrimonia pilosa ledeb. for treating or preventing diabetic complication, anti-oxidative effect and improving lipid metabolism - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for antioxidant, lipid metabolism improvement and prevention and treatment of complications caused by diabetes, including Seonchocho extract, the sunchocho extract of the present invention reduces the concentration of plasma total cholesterol and triglycerides, excellent antioxidant capacity It can be used to improve the lipid metabolism and prevent and treat diabetic complications due to oxidative stress and medicines and functional foods.

Complications from Diabetes, Oxidative Stress, Seonhakchocho Extract

Description

Pharmaceutical composition comprising the extract of Agrimonia pilosa Ledeb. for treating or preventing diabetic complication, anti-oxidative effect and improving lipid metabolism}             

1 is a graph of the antioxidant activity of L-ascorbic acid as a sunchocho extract and a control by the hydrogen donating ability by DPPH (1,1-diphenyl picrylhydrazyl) method,

Figure 2 is a result of measuring the TBARS (thiobarbituric acid-reactive substances) levels of the diabetic animal model in the control group and Seonhakcho extract administration group,

Figure 3 is a result of measuring the superoxide dismutase (SOD) levels of the diabetic animal model in the control group and Seonhakcho extract administration group,

4 is a result of measuring the catalase (catalase) level of the diabetic animal model in the control group and Seonhakcho extract administration group,

5 is a result of measuring the plasma triglyceride and total cholesterol concentration of the diabetic animal model in the control group and Seonhakcho extract administration group.

The present invention relates to a pharmaceutical composition containing an antioxidant effect, Agrimonia pilos a Ledeb. Extract as an active ingredient, improving lipid metabolism and preventing and treating complications caused by diabetes.

Diabetes mellitus is a disease in which hyperglycemia occurs because cells in the body cannot use sugar due to reduced insulin secretion or reduced insulin function. In diabetes, hormonal imbalances, including insulin, are characteristic of hyperglycemia due to abnormal physiological metabolic control functions such as carbohydrate-containing protein, lipid, and electrolyte metabolism (Gonuth, SM, Ann. Intern. Med. , 79 , pp812-822, 1973) The persistence of these hyperglycemia symptoms can lead to blood circulation disorders, retinal damage, nerve cell damage, renal failure, and vascular complications. And serious chronic complications (Kannel, WB and McGee, DL JAMA, 241 , pp 2035-2038, 1979).

In particular, the incidence of cardiovascular diseases such as atherosclerosis, cerebral infarction, cerebral thrombosis and myocardial infarction is higher in diabetic patients than normal patients (Fuller, JH, Lancet, 1 , pp1373-1376, 1980). Diabetes patients have a high risk of death from coronary artery disease or cerebrovascular disease, which are often caused by hypertension, hyperlipidemia, obesity, etc. (Hu-Beom Bum. The Korean Nutrition Society. Abstract, pp. 15-18, 1984). It has been reported that 67% of people with type 2 diabetes have more than one or more types of lipid metabolism (Harris, MI Diabetes Care , 23 , pp754-758, 2000). Patients with type 2 diabetes have abnormal lipid metabolism with increased triglyceride and cholesterol levels and decreased HDL-cholesterol (Goldberg, RB Diabetes Care, 4 , pp561-572, 1981), which is a complication of diabetic coronary artery disease. (Reaven, JW Am. J. Med., 83 , pp 31-40, 1987).

Diabetes complications are known to be caused by elevated blood glucose levels and free radicals caused by diabetes (Kazuyuki Hayashi, Masakazu Haneda, Daisuke Koya et al. Diabetes Res. Clin. Pract., 52 , pp85-96, 2001: Joshua P Klein and Stephen G Waxman.The Lancet Neurology , 2 (9) , pp548-554, 2003). In the diabetic state, the production of free radicals is increased by hyperglycemia such as glucose autooxidation and non-enzymatic protein glycosylation, which is very susceptible to oxidative damage caused by various free radicals. The Korean Diabetes Association ( 1998, Korea Medicine). Hydrogen peroxide produced under normal conditions can be removed by intracellular antioxidant systems and antioxidant enzymes, but lipid peroxidation is increased in abnormal conditions such as diabetes (Wills ED Biochem J. , 99 , pp667-675, 1965). Shweta Bhatiaa, et al., Clinical Biochemistry , 36 , pp 557-562, 2003). Sustained hyperglycemia during diabetes can increase the production of oxygen free radicals through autooxidation and glycosylation of non-enzymatic proteins.

In patients with diabetes, free radicals are one of the major mechanisms involved in the development of chronic complications. Diabetes patients are prone to free radical production, which increases the level of lipid peroxidation in liver tissue, heart muscle and serum, and is likely to cause cardiovascular diseases such as stroke and myocardial infarction (Joshua P Klein and Stephen G Waxman, The Lancet Neurology , 2 (9) , pp 548-554, 2003). In diabetic patients, the incidence of vascular diseases such as coronary heart disease and atherosclerosis is known to be 2 to 6 times higher than that of non-diabetic patients. Cardiac circulatory complications are the leading cause of death in diabetic patients. In addition, diabetic eyes, like other organs, are susceptible to metabolic disorders caused by diabetes. Increased non-enzymatic glycosylation, oxidation of proteins and lipids, and reduced glutathione in diabetes are harmful substances. It reduces the skin's resistance to and causes clouding and cataracts of the lens (Yarat A., et al., Free Rad. Biol. Med., 31 (9) , pp 1038-1042, 2001).

Agrimonia pilos a Ledeb. Is a perennial herb from the Rosaceae family. Its ingredients include agrimonin, agrimonolide, tannin, sterol, organic acid, saponin, and astringent hemostasis and ripening core. It is known to have the effect of 强 心), branch office, hemostasis, and health (健胃) (Bo Bo-seop and Shin Mingyo, Hyangjedae, Yeonglimsa, pp 636-637, 1998). In addition, researches on anti-aging cosmetics (Japanese Patent No. 10,226,617), anti-cancer effect ( ACS Symp. Ser. 662 , pp245-259, 1997), whitening effect (Japanese Patent No. 8,175,957) containing sunhakcho extract Although it has been reported, none of the above documents discloses or teaches the anti-oxidant effect of Seonhakchocho extract and the prevention or treatment of diabetes complications by improving lipid metabolism.

Accordingly, the present inventors prepared the Seonhakcho extract having excellent antioxidant activity, and as a result of measuring blood lipid composition and antioxidant enzyme activity, the present invention was completed by confirming that the lipid metabolism improvement and antioxidant activity were excellent in improving diabetes complications. It was.

It is an object of the present invention to provide a composition for antioxidant, lipid metabolism improvement and prevention and treatment of diabetes complications containing sunhakcho extract having antioxidant activity as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for improving lipid metabolism and preventing and treating complications of diabetes containing Agrimonia pilos a Ledeb. Extract as an active ingredient.

The present invention also provides an antioxidant pharmaceutical composition containing Seonchocho extract as an active ingredient.

The lipid metabolism includes metabolism of blood cholesterol and triglyceride in blood.

In addition, the complications of diabetes mellitus and cataracts, skin symptoms caused by coronary heart disease, arteriosclerosis, cardiovascular diseases such as myocardial infarction, skin infections, microangiopathy, neuropathy, skin reactions with therapeutic drugs, cataracts, diabetes Diseases such as retinopathy, diabetic nephropathy and diabetic neuropathy.

The extract includes water extracted from C ₁ to C ₄ lower alcohols such as water, ethanol, methanol, or a mixed solvent thereof.

Hereinafter, the present invention will be described in detail.

Seonchocho extract of the present invention can be obtained as follows.

By purchasing seonhakcho of the present invention, freeze-dried by grinding about 2 to 15 times the seonhakcho sample weight, preferably from C ₁ to C _4, such as about 5 to 10 times by volume up to the water and methanol, ethanol, butanol, A lower alcohol polar solvent or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably 70 to 100% methanol at room temperature for about 1 hour to 1 day, preferably 6 hours to Using extraction methods such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for 12 hours, preferably hot water extraction to separate the extract from the residue by filtration under reduced pressure, and then the obtained residue at 300 seconds at the start of extraction In polar solvents of water and C ₁ to C ₄ lower alcohols such as methanol, ethanol, butanol and the like, or about 1: 0.1 thereof, in volumes of water up to about 2 to 10 times, preferably about 3 to 5 times, by weight A mixed solvent having a mixing ratio of 1 to 10, preferably 70 to 100% methanol using an extraction method such as hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction for about 1 to 5 hours at room temperature Preferably, the hydrothermal extract and then filtered under reduced pressure and concentrated to obtain the extract of Seonhakcho.

In addition, conventional fractionation processes can also be carried out (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis, 3rd Ed. , Pp 6-7, 1998).

The present invention provides a pharmaceutical composition for the improvement of lipid metabolism and the prevention and treatment of diabetes complications containing the Seonhakcho extract obtained by the above manufacturing process as an active ingredient.

The present invention also provides an antioxidant pharmaceutical composition containing the Seonhakcho extract obtained by the manufacturing process as an active ingredient.

Seonhakcho extract obtained by the above method significantly reduces the total cholesterol concentration and the concentration of triglycerides in the plasma bar shows an improvement effect of lipid metabolism.

The pharmaceutical composition for improving the antioxidant effect, lipid metabolism and prevention and treatment of diabetic complications of the present invention comprises 0.1 to 50% by weight of the Seonchocho extract based on the total weight of the composition.

The pharmaceutical composition comprising Seonchocho extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of Seonchocho extract of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

Pharmaceutical compositions comprising Seonhakcho extract according to the present invention, respectively, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions according to a conventional method. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising wood vinegar include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations 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. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of Seonchocho extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, Sunchocho extract of the present invention is preferably administered at 0.0001 to 100mg / kg, preferably at 0.001 to 100mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

Seonchocho extract of the present invention can be administered to various mammals such as mice, mice, livestock, humans. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a health functional food comprising the Seonhakcho extract and food acceptable food supplements exhibiting the effects of antioxidant effect, improvement of lipid metabolism and prevention and treatment of diabetes complications. Examples of the food to which the Seonhakcho extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to foods or beverages for the purpose of antioxidant effects, lipid metabolism improvement and prevention and treatment of diabetes complications. At this time, the amount of the Seonchocho extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5g, preferably 0.3 to 1g based on 100ml have.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the Seonhakcho as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. 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 (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates 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, Seonhakcho extract of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, such as flavoring agents, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the sunhakcho natural fruit juice of the present invention and may contain a fruit flesh for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of suncho vinegar of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Example 1. Preparation of Seonhakchocho extract

Sunhakcho of the present invention was used in the form of dried leaves from Kwangmyeong Herbal Medicine Co., Ltd. (Korea) was added to 7g of 95% methanol to 200g of sunhakchocho extract for 24 hours, and then filtered under reduced pressure with a filter paper (Watman, USA) Then, the filtrate was collected and concentrated under reduced pressure at 37 ° C. using a vacuum rotary concentrator to obtain 29.6 g of methanol extract of Sunhakcho, which was used for experiments while storing at −20 ° C.

Experimental Example 1. In vitro ( In vitro Of antioxidant activity

Seonhakcho vitro (In vitro) method DPPH (1,1-diphenyl picrylhydrazyl) for measuring the antioxidant activity (Blois, MS: Antioxidant determination by the use of a stable free radical, Nature, 26, 1199-1200, 1958) the Was used.

The DPPH method is reduced by physiologically active substances that exhibit antioxidant activities such as tocopherol, ascorbate, flavonoid compounds, aromatic amines, Maillard browning products, and peptides. It is a method to measure the antioxidant effect by hydrogen donating ability according to the degree of dark purple discoloration. In this case, L-ascorbic acid was used as a control, and 2 x 10 ^-4 M DPPH was added to each sample of Seonchocho methanol extract obtained in the above example, and the absorbance was measured at 517 nm. The results are shown in Table 1.

Free radical scavenging capacity according to treatment concentration (%) Treatment concentration (mg / ml) L-ascorbic acid Seonhakcho methanol extract 0.5 92.0 80.5

As a result, Seonchocho methanol extract showed 80.5% at a concentration of 0.5 mg / ml, and L-ascorbic acid used as a control showed 92.0% free radical scavenging ability ( FIG. 1 ). That is, since cell damage caused by free radicals is known to be closely related to the occurrence of diabetes mellitus and complications of diabetes mellitus, it can be confirmed that Methanol extract of Sunhakcho, which exhibits a strong antioxidant effect, may show an improvement in diabetes complications.

Reference Example 1. Preparation of Type 2 Diabetes Animal Model and Sample

1-1. Laboratory Animals and Diet

In order to measure the effects of in vitro diabetes and diabetic complications of Sunhakcho, which were shown to have excellent in vitro antioxidant activity in Experimental Example 1, four-week-old db / db mice, the type 2 diabetes animal model, were measured. The control group was separated by the ovary method to have similar mean blood glucose levels, and the control group was treated with AIN-93G basal diet (see Table 2 below), and the group containing the Seonchocho extract extract containing 0.5% lyophilized Seonchocho for 6 weeks. Ingested. The diet and drinking water were allowed to be ad libitum freely. The temperature and humidity of the feeding room were maintained at 20 ~ 25 ℃ and 50 ~ 60%, respectively, and the lighting was turned on and off every 12 hours. Animal weight and dietary intake were measured three times a week.

Dietary composition of the control group Furtherance Control Diet (%) Cornstarch ¹⁾ 39.7 Casein ²⁾ 20.0 Dextrinized starch 13.2 Sucrose ³⁾ 10.0 α-cellulose ⁴⁾ 5.0 Mineral mixtures ⁵⁾ 3.5 Vitamin mixtures ⁵⁾ 1.0 L-cystine ⁵⁾ 0.3 Choline bitartrate ³⁾ 0.25 Tert-Butylhydroquinone 0.0014 Soybean oil ⁶⁾ 7.0 ¹⁾ Target company, Korea ²⁾ Murry goulburn Co., Australia ³⁾ Samyang, Korea ⁴⁾ Sigma, USA ⁵⁾ ICN Biochemical, USA ⁶⁾ CheilJedang, Korea

As a result of measuring body weight and dietary intake, the control group and Sunhakcho group weighed 38.2 ± 2.4g and 37.5 ± 4.1g, respectively. 4.9 ± 0.3 g / day, 5.1 ± 0.6 g / day did not show a significant difference (see Table 3 below).

Body weight and dietary intake of control group and sunhakcho group in diabetic animal model Control Three hundred seconds Body weight (g) ¹⁾ 38.2 ± 2.4 ²⁾ 37.5 ± 4.1 Dietary Intake (g / day) 4.9 ± 0.3 5.1 ± 0.6 ¹⁾ Weight after 6 weeks of dietary intake ²⁾ Mean ± SD

1-2. Sampling

Animals were sacrificed at 6 weeks of diet. The animals were fasted for 14 hours before sacrifice and suffocated with carbon dioxide gas and blood was collected from the heart with a syringe containing 10 mg of EDTA (ehylene diamine tetra acetic acid). Blood was centrifuged at 3,000 rpm for 15 minutes to collect plasma and stored at -70 ° C. Liver tissue was extracted and stored at -70 ° C.

1-3. Statistical analysis

All results were expressed as mean ± standard deviation, and significance test between control group and Seonchocho extract administration group was performed using Student's t-test (α = 0.05).

Experimental Example 2 Determination of Lipid Peroxidation Levels of Sunhakcho Extract

2-1. TBARS measurements

The measurement of liver TBARS (thiobarbituric acid-reactive substances) content was measured using thiobarbituric acid (Ohkawa H., Ohisi N., and Yagi K. Anal Biochem , 95 , pp351, 1979). The content of malondialdehyde (MDA) reacting with TBA) was measured, and 1,1,3,3, -tetramethoxypropane (TMP) was used as a control. In addition, the protein content of the homogenate was measured using the modified Lowry's method (Marklund S. and Marklund G. Eur J Biochem , 47 , pp469, 1974). TBARS content in the liver is shown in FIG. 2 .

As a result, the TBARS content (0.94 ± 0.21 nmol MDA / mg protein) of liver tissues of Sunhakcho group was shown as the control (1.27 ± 0.27 nmol MDA / mg protein). In other words, ingestion of Seonhakcho extract may reduce the oxidative stress of diabetic rats by inhibiting lipid peroxidation of liver, thereby contributing to the prevention of tissue damage and diabetic complications caused by diabetes.

2-2. Superoxide dismutase (SOD) activity measurement

SOD activity was measured at 420 nm using the method of Marklund S. and Marklund G. Eur J Biochem , 47, pp469, 1974 using color development by automatic oxidation of pyrogallol in alkaline state. . SOD 1 unit (unit) was the amount of enzyme required to inhibit the autoxidation of galrol blood for 1 minute and up to 50%, the results are shown in Fig.

As a result, the total SOD activity of the liver was significantly higher than that of the control group in 11.3 ± 1.03 U / mg protein and 8.13 ± 0.59 U / mg protein in the control group (P <0.05). In other words, the decrease in lipid peroxide concentration of liver tissues of Sunhakcho group and the increase of SOD activity that primarily breaks down free radicals into hydrogen peroxide (H ₂ O ₂ ) indicate that the active ingredient of Sunhakchocho extract can protect diabetic animals from peroxidative damage. Indicates that it can.

2-3. Catalase Activity Measurement

Catalase converts cytotoxic hydrogen peroxide back into oxygen and water to protect the body from peroxidation (MM Kesavulu, B. Kameswara Rao, R. Giri, J. Vijaya, G. Subramanyam and Ch. Apparao) Diabetes Research and Clinical Practice , 53 (1) , pp 33-39, 2001).

10 times cold 50 mM phosphate buffer (pH 7.4) was added to the liver tissue and homogenized with a glass teflon homogenizer. Cellular fractions obtained by centrifugation of the homogenate at 3,000 rpm for 10 minutes were used as samples for catalase enzyme measurement. Catalase activity was measured by Aebi (Aebi H. Catalase in vitro in method in enzymology, Academic press, Grlando, Florida, 105, pp 121, 1984). Enzyme activity was 1 unit of the amount of enzyme to decompose 1u mole substrate for 1 minute, the results are shown in FIG.

As a result, hepatic catalase activity was 5.827 ± 1.329 U / mg mg in the control group and 8.466 ± 1.167 U / mg mg in the control group, but there was no significant difference.

Experimental Example 3. Lipid metabolism improvement effect of Sunhakcho extract

3-1. Plasma Triglycerides and Total Cholesterol

Triglycerides in plasma were measured by Buocole's method (Buoccole, G. and David, H., Clin. Chem., 19 , pp476, 1973).

Specifically, the triglyceride by the lipase is decomposed into glycerol and fatty acid glycerol is again changed glycerol-1-phosphate glycerol-1-phosphate is cleaved by the GPO is H ₂ O ₂ is generated, the generated H ₂ O ₂ Is reacted with aminoatiphyrin and TOOS to produce red quinone by oxidative condensation reaction, and then it is reacted for 5 minutes at 37 ℃ using the method of quantifying triglyceride by colorizing and measuring the absorbance at 546 nm. Quantification by

As a result, the plasma triglyceride concentration was 181.1 ± 10.3 mg / dL in the control group and 150.2 ± 10.7 mg / dL in the Seonhakcho group, and the long-term intake of the three hundred sec extract significantly decreased the concentration of triglyceride in diabetic animals. It was confirmed (P <0.05).

Meanwhile, plasma total cholesterol was measured by Saleh's method (Sale, FO, et al., Anal. Biochem. , 142 , pp347-348, 1984).

Specifically, there are free and ester type cholesterol in serum, and when the ester type hydrolyse reacts with cholesterol ester hydrolase, it is decomposed into free type and fatty acid, and the free type reacts cholesterol oxidase with H ₂ O ₂ and δ. Cholestenone is produced. When the peroxide enzyme, 4-aminoantipyrine and phenol are simultaneously reacted with the produced H ₂ O ₂ , quinone is produced by oxidative condensation reaction. Absorbance was measured and quantified by sample and standard.

As a result, the total cholesterol concentration was 203.5 ± 10.0 mg / dL in the control group and 128.0 ± 4.9 mg / dL in the Seonhakcho group. Long-term intake of Seonchochocho extract significantly reduced plasma cholesterol levels in diabetic animals. It could be confirmed (P <0.001) (see FIG. 5 ).

Experimental Example 4. Acute Toxicity Test

Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats. Two animals of each group were orally administered three hundred seconds of the present invention at a dose of 100 mg / kg. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to visually observe abnormalities in organs and thoracic organs.

As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings. As a result, Seonhakcho of the present invention did not show a change in toxicity even in rats up to 100 mg / kg, respectively, the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance more than 100 mg / kg.

The preparation example of the pharmaceutical composition containing Seonchocho extract of the present invention will be described, but the present invention is not intended to limit the present invention but is intended to be described in detail.

Formulation Example 1 Preparation of Powder

Seonhakcho methanol extract powder 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Seonchocho methanol extract powder 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Seonchocho methanol extract powder 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Seonchocho methanol extract powder 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ ㅇ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Seonhakcho methanol extract powder 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

Seonchocho methanol extract powder 1000mg

Vitamin Mixture

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic 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 mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

Seonchocho methanol extract powder 100mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantity

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, Seonhakchocho extract of the present invention is excellent in inhibiting lipid peroxidation, and by reducing oxidative stress, diabetes, such as stroke, cardiovascular diseases such as myocardial infarction, diabetic eye disease caused by oxidative stress in diabetic patients It can be used in medicines and health foods for the prevention and treatment of complications.

Claims (7)

A pharmaceutical composition for improving lipid metabolism and preventing and treating complications caused by diabetes mellitus, which contains Agrimonia pilos a Ledeb. Extract as an active ingredient. A pharmaceutical composition for antioxidant, containing Seonchocho extract as an active ingredient. The pharmaceutical composition according to claim 1 or 2, wherein the Seonchocho extract is extracted with water and C ₁ to C ₄ lower alcohols such as methanol and ethanol or a mixed solvent thereof. The pharmaceutical composition of claim 1 or 2, wherein the Seonchocho extract comprises 0.1-50% by weight of Seonchocho extract based on the total weight of the composition. According to claim 1 or 2, wherein the complications caused by diabetes mellitus cardiovascular diseases, such as coronary heart disease, arteriosclerosis, myocardial infarction, skin infections, microangiopathy, neuropathy, skin reactions due to therapeutic drugs, etc. At least one pharmaceutical composition selected from the group consisting of clouding of symptoms and lens and cataract, diabetic retinopathy, diabetic nephropathy, diabetic neuropathy. A dietary supplement comprising Seonhakchocho extract, which exhibits an antioxidant effect, improved lipid metabolism, and prevents complications from diabetes, and a food acceptable additive. The health functional food of Claim 6 which is a health drink.

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