KR101402929B1 - Pharmaceutical composition and functional food for prevention or treatment of acute renal failure comprising a herbal extract - Google Patents

Abstract

The present invention relates to a natural product extract effective for preventing or treating acute renal failure, a pharmaceutical composition for preventing or treating acute renal failure containing an overprize extract or a fraction thereof as an active ingredient, and a health functional food composition for improvement.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition and a functional food for preventing or treating acute renal failure containing an extract of a natural product as an active ingredient and a functional food for prevention or treatment of acute renal failure comprising a herbal extract,

The present invention relates to a pharmaceutical composition and a health functional food for preventing or treating acute renal failure containing an extract of Trichosanthis semen or a fraction thereof as an active ingredient.

Acute renal failure is a clinical syndrome that causes rapid renal dysfunction due to various causes such as decreased renal blood flow, glomerulonephritis, nephrotoxic antibiotics, and the use of anticancer drugs. It is associated with decreased glomerular filtration rate (GFR) Decrease in urine volume, hyperglycemia due to the accumulation of nitrogen waste, and imbalance of body fluids and electrolytes. The renal dysfunction of acute renal failure is a high-risk disease with a mortality rate of 50% due to the difficulty in recovery due to the failure of initial cause removal, and the clinical efficacy of the newly proven therapies despite the ongoing research and efforts This is not yet clear.

Acute renal failure has symptoms of hyperlipidemia, called uremia, which means that kidneys are suddenly lost due to damage to tubules or decreased glomerular filtration rate. Healthy adults who require a normal diet should excrete a minimum of about 400 ml of urine per day to excrete waste products in the body through the kidneys. Adults who have lost renal function excrete less than 400 ml of urine. The causes of acute renal failure can be divided into three major categories based on the kidneys. Renal failure caused by obstruction of the blood flow to the kidneys, renal insufficiency, which is a problem of the renal itself, and kidney failure caused by obstruction of any part of the urinary tract from the tubule to the urethra It is classified as New Hope. Symptoms include severe dehydration, excessive bleeding, burns, severe vomiting, diarrhea, pancreatitis or peritonitis, and sepsis. Renal excretion of the kidneys is increased, and blood flow to the kidneys is reduced when diabetes occurs, which is called renal failure. In addition, when a change in the kidney parenchyma due to a direct damage of the kidney itself due to a disease or a neoplastic cytotoxic substance is present, the disease is called glomerulonephritis, including glomerulonephritis, kidney microangiopathy, prolonged systemic renal failure, Renal tubular necrosis, thrombosis, trauma, atherosclerosis, and tumors within the kidney.

As mentioned above, there are various causes but it is not necessarily one cause, but various causes can cause mutual induction of multiple or other causes to cause renal failure, which is caused by anorexia, nausea and vomiting, resin progression, edema and hypertension If it gets worse, it can lead to severe cases of breathing difficulty, convulsions, coma and eventually death.

Cisplatin (cis-diaminedichloroplatinum II), which is used for the induction of acute renal failure, is one of the commonly used anticancer drugs and causes acute renal failure due to structural abnormality of the tubule. These cisplatin - induced tubular injuries are known to play a major role in free radicals. The increase in lipid peroxidation in the kidney is also involved in the renal toxicity by cisplatin, and it is known that cisplatin itself suppresses the antioxidant effect in the kidney, thereby reducing the content of glutathione (GSH) in the kidney. Although the exact pathophysiology of acute renal failure due to cisplatin has not yet been fully understood, it has been reported that the mechanism involved in chemotherapy is associated with environmental factors.

The cisplatin-induced acute renal failure model is now one of the most commonly used acute renal failure animal models. Captopril, administered to the control group, inhibits the angiotensin converting enzyme (ACE), an essential enzyme that forms angiotensin II, and is known as a representative hypertension treatment for lowering blood pressure. Angiotensin II is an enzyme that increases blood pressure by causing arteriovenous shrinkage of the body, and captopril suppresses the formation of it and lowers blood pressure. In general, ACE inhibitors are known to be effective not only for hypertension due to the heart, but also for kidney diseases which are closely related to hypertension. Therefore, ACE inhibitors are currently being used as a reference drug in the development of a treatment for renal failure, The effect of antioxidant treatment on renal failure is well known (El-Sayed el-SM et al., 2008, Takako Yokozawa, et al., 2000).

Recently, it has been reported that BUN, creatinine, and weight change are effective in acute renal failure even when natural products are used for acute renal failure (Takayama et al., 2008; Takako Yokozawa, et al. al 2000). In the assessment of the severity of acute renal failure, it is common to measure the levels of "BUN (Blood Urea Nitrogen), Blood Creatinine, and Weight Change", which induce symptoms and exacerbate symptoms. The BUN level is a blood biochemical indicator of the blood level of urea nitrogen, the metabolic product of proteolysis. The elevation of BUN in the blood usually indicates the presence of kidney disease. In addition, creatinine is a nitrogen product formed by non-protein-derived muscle metabolism. Increasing creatinine content in blood together with BUN implies a decrease in glomerular filtration rate (GFR). Changes in BUN and creatinine levels in cisplatin-induced acute renal failure have been used as a basic indicator of cisplatin-induced acute renal failure and have been shown to inhibit cisplatin-induced weight loss in a dose- It was recognized as effective. The decrease in body weight and body weight change induced by cisplatin is judged to be a secondary change accompanied by direct toxicity of cisplatin itself or acute renal failure. To date, such a change in body weight is the most basic indicator .

Accordingly, the present inventors have conducted intensive studies on the natural medicines for preventing or treating acute renal failure, such as the above-mentioned extracts and their fractions in vitro in vitro ) and in vivo experiments ( in vivo studies have demonstrated efficacy in the prevention or treatment of acute renal failure and completed the present invention.

It is an object of the present invention to provide a pharmaceutical composition or a health functional food containing an overprual extract or a fraction thereof as an active ingredient capable of preventing or treating acute renal failure.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating acute renal failure, which comprises an overprual extract or a fraction thereof as an active ingredient.

The term " over phosphorus " as used in the present invention means that the ripe seeds belonging to the bark and belonging to the genus Trichosanthes kirilowii Maxim. Or T. rosthornii Harms are dried. Influenza is a febrile bronchitis (a person with a flank in the side) in one room, and it is good for a patient with a lot of sputum. It is known to have good pharmacological effect on chest pain which feels acute bronchitis, pleuritis, pneumonia caused by cough, sputum and stiffness. In addition, it is used as a medicine to cough better, and it has the effect of easily improving the constipation symptoms of people with digestive problems or gastrointestinal disorders that can not digest the food even with small amount of stress due to habitual constipation and nervousness.

The term "extract " as used in the present invention means a specific ingredient extracted from the over-the-fore by using a liquid solvent. As the solvent, water, C _1-4 alcohol or a mixture thereof may be used. In order to efficiently extract the active ingredient, it is preferable to extract it with methanol or ethanol, and it is preferable to use 70% ethanol or methanol, but it is not limited thereto.

The overrual extract of the present invention may be any one selected from the group consisting of ultrasonic extraction, hot water extraction, cold extraction, reflux cooling extraction, and steam extraction. It is preferable to use an ultrasonic extraction method in order to efficiently extract an effective ingredient.

Preferably, the fraction of the over-extracted extract is a hexane fraction, an ethyl acetate fraction, a butanol fraction or a water fraction.

As used herein, the term "acute renal failure" refers to a clinical syndrome that results in acute renal failure resulting from a variety of causes including acute renal failure, glomerulonephritis, nephrotoxic antibiotics, and the use of anticancer drugs. The perilla extract or the fraction of the over-the-top extract according to the present invention can be usefully used for preventing or treating such diseases.

On the other hand, cisplatin is a chemotherapeutic agent for malignant tumors including a variety of cancers including testis, head and neck, ovary, cervix, non-small cell, and kidney (Pala and Dong, 2008; Wang and Lippard, 2005). However, 25-35% of cancer patients experience a sudden decrease in renal function after a single dose of cisplatin (Luke et al., 1992), and the greatest side effect of the kidney function is a sudden decrease in renal function, (Ries and Klastersky, 1986).

In the present invention, it has been confirmed that a composition containing an overprual extract or a fraction thereof is effective for preventing or treating acute renal failure. According to one embodiment of the present invention, it was confirmed that the effect of changing body weight and BUN, creatinine, which are indicators of acute renal failure therapy, is effective for preventing or treating acute renal failure.

As used herein, the term "prevention" means any action that inhibits or delays disease by administration of a composition comprising the over-the-fruit extract or its fractions. The term "treatment ", as used herein, refers to any action that improves or alleviates symptoms of acute renal failure by administering a composition comprising the over-the-top extract or its fractions.

The composition of the present invention may contain, for administration, a pharmaceutically acceptable carrier, excipient or diluent in addition to the above-described effective ingredients. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The composition of the present invention may be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository or sterilized injection solution according to a conventional method. In detail, when formulating, it can be prepared by using diluents or excipients such as fillers, weighing agents, binders, humectants, disintegrants, surfactants and the like which are generally used. Solid formulations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation can be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, gelatin, and the like in the overpouring extract. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration, liquid paraffin, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and tasks. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. As a base for suppositories, it is possible to use witepsol, macrosole, tween 61, cacao paper, laurin, glycerogelatin and the like.

The composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the desired method, and the dose may be determined depending on the condition and weight of the patient, The mode of administration, the route of administration, and the time, but may be suitably selected by those skilled in the art. The daily dose of the over-the-top extract or its fraction is preferably 1 mg / kg to 500 mg / kg, and may be administered once a day to several times per day as needed.

The present invention also provides a health functional food for preventing or ameliorating acute renal failure containing an overprual extract or a fraction thereof as an active ingredient.

The health functional food may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, alginic acid and its salts, , Protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, it may contain natural fruit juice and fruit juice and pulp for the production of commercial beverages. These components may be used independently or in combination. The health functional food may be in the form of any one of meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, .

In addition, the health functional food may further include food additives, and the suitability of the food functional food as a "food additive " is not limited to those specified in the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the products that have been used in the above-mentioned "food additives" include natural products such as ketones, chemical products such as glycine, potassium citrate, nicotinic acid and cinnamic acid, sensory coloring matter, licorice extract, crystalline cellulose, high- - Mixed preparations such as a sodium glutamate preparation, a noodle-added alkaline agent, a preservative preparation, a tar coloring agent and the like.

At this time, the extract according to the present invention, which is added to foods containing beverages in the process of manufacturing a health functional food, can be appropriately increased or decreased as required, and preferably 1 to 15% by weight based on 100% Is preferably added.

The present invention is a natural extract effective for the prevention or treatment of acute renal failure, and can be used not only as a pharmaceutical composition for preventing or treating acute renal failure, but also as a health functional food.

In addition, since it has an effect of inhibiting side effects induced by anticancer drugs such as cisplatin, it can be effectively used as an inhibitor of side effects of anticancer drugs.

Brief Description of the Drawings Fig. 1 is a graph showing the results of a comparison between the (a) perineal extract, (b) the hexane fraction of the perineum extract, (c) the ethyl acetate fraction of the perineum extract, (d) the butanol fraction of the perinein extract, (e) MTT assay result showing cell viability of the water fraction of the over-extracted extract.
FIG. 2 is a graph showing the cytotoxicity induced by Cisplatin in accordance with an embodiment of the present invention. FIG. 2 (a) shows the extract of peruvirus, (b) the hexane fraction (HA) (EA), (d) the butanol fraction (BU) of the perilla extract, and (e) the water fraction (WT) of the perilla extract.
FIG. 3 is a graph showing the results of cell viability measurement of a fractionated drug by cisplatin treatment according to an embodiment of the present invention. ), (c) the ethyl acetate fraction (EA) of the perilla extract, (d) the butanol fraction (BU) of the perlein extract and (e) the water fraction (WT) of the perlein extract will be.
(B) the ethyl acetate fraction of the perlein extract; (c) the butanol fraction of the perlein extract; and (d) the butanol fraction of the perlein extract. The results of measurement of ROS (Reactive Oxygen Species) production rate of water fraction of perilla extract.
Figure 5 is a graph showing the effect of the extract of perilla (a), the fraction of hexane (HA), (c), the ethyl acetate fraction (EA) (BU) and (e) the water fraction (WT) of the extract of perineum.
FIG. 6 is a graph showing the results of an animal test according to an embodiment of the present invention, in which a change in body weight was measured after administering an extract of perianths.
FIG. 7 shows the result of measurement of BUN change after administering an extract of perennials as an animal test result according to an embodiment of the present invention.
FIG. 8 is a graph showing the results of creatinine changes measured after administering an extract of perianths as an animal test result according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following Production Examples and Examples. However, the following Preparation Examples and Examples are for illustrating the present invention, but the scope of the present invention is not limited thereto.

Example  One: Superfluous  Extract preparation

3.0 kg of pulverized pulverized product obtained by purchasing commercially available pulp was placed in 30 L of 70% ethanol and ultrasonically extracted three times for 1 hour using an ultrasonic extractor. The extract was filtered through a Wetman (Whatman 46 x 57 cm) filter paper to remove insoluble matters and then concentrated under reduced pressure at 60 ° C with a condenser equipped with a condenser. To completely remove the solvent of the concentrated extract, 100 ml of purified water was added to the suspension to obtain 104.39 g of an extract (yield: 3.48%) by freeze-drying.

Example  2: Superfluous  Extract Fraction  Produce

1) Preparation of over-phosphorus extract

30 g of 70% ethanol was added to 300 g of the pulverized pulp obtained by purchasing commercially available pulp, and ultrasonication was performed three times for 1 hour using an ultrasonic wave extractor. The extract was filtered through a Whatman (46 × 57 cm) filter paper to remove insoluble matter, and then concentrated under reduced pressure at 60 ° C. with a condenser equipped with a condenser. To completely remove the solvent of the concentrated extract, 100 ml of purified water was added to the suspension, and 17.55 g of the extract was obtained by using a freeze dryer (yield: 5.85%).

2) Preparation of hexane fraction

17 g of the perilla extract prepared in Example 2-1) was suspended in 0.4 L of distilled water and 0.4 L of hexane was added thereto to carry out the solvent fraction three times. The hexane fraction was recovered, concentrated under reduced pressure and lyophilized to obtain 2.26 g of hexane fraction (yield from the over-phosphorus extract of Example 1: 13.29%).

3) Preparation of ethyl acetate fraction

After hexane fractionation of the above Example 2-2), 0.4 L of ethyl acetate was added to the remaining water layer, and the solvent fraction was applied three times. The ethyl acetate fraction was collected, concentrated under reduced pressure, and lyophilized to obtain 0.93 g of an ethyl acetate fraction (yield: 5.46% from the over-phosphorus extract of Example 2-1).

4) Production of butanol fraction

After the ethyl acetate fraction of Example 2-3), 0.4 L of butanol was added to the remaining water layer to perform the solvent fraction three times. The butanol fraction was recovered, concentrated under reduced pressure and lyophilized to obtain 1.61 g of butanol fraction (yield: 9.44% from the over-phosphorus extract of Example 2-1).

5) Preparation of water fractions

The remaining water layer after the butanol fraction of Example 2-4) was concentrated under reduced pressure and lyophilized to obtain 10 g of a water fraction (yield from the over-phosphorus extract of Example 2-1): 58.82%).

Experimental Example  1: Cytotoxicity experiment

The cytotoxicity of the perilla extract prepared in Example 1 and the fraction of perilla extract prepared in Example 2 (hexane, ethyl acetate, butanol and water fraction) was analyzed as follows.

A) Experimental method

Porcine renal epithelial cells were plated at 1 × 10 ⁴ cells / well in a 96-well plate, and then the extracts of the perlite extract prepared in Example 1 and the extracts of perlite extract prepared in Example 2 Hexane, ethyl acetate, butanol and water fractions) were treated at a concentration of 10/20/50/100/200 μg / ml (treatment concentration: 20/40/100/200/400 μg / ml). After treatment with 50 μl of the drug, the final concentration of PBS was adjusted to 1% (treatment concentration: 2%) and 20 mg / ml (PBS: 100 mg / And the mean value of each well was used.

B) Experimental results

The cytotoxicity test results are shown in Fig. As shown in FIG. 1, the results of 24-hour experiments were compared with each other, and it was confirmed that the fractions of perilla extract and perilla extract (hexane, ethyl acetate, butanol and water fractions) there was.

Experimental Example  2: in vitro  Acute renal failure efficacy experiment

1) MTT Assay

MTT assay experiments were performed to measure cell activation in the kidney cells of the perilla extract and the fraction of perilla extract (hexane, ethyl acetate, butanol and water fractions).

A) Experimental method

Porcine renal epithelial cells were plated in 96-well plates at 1 × 10 ⁴ cells / well. Based on the toxicity test result of the drug itself after 24 hours, the drug which was judged as non-toxic drug was prepared by diluting to the three final concentrations according to the drug concentration determination experiment. The drug was treated with 50 ㎕ of each concentration and the drug treatment After 2 hours, cisplatin was added to a final concentration of 15 占 퐂 / ml and cultured for 24 hours. After 24 hours, the assay reagent of EZ-Cytox Cell Viability Assay Kit was added to each well with 10 μl of each well to prevent foaming by using Micro Multi Pippet, and the absorbance was measured at 450 nm by reacting for 4 hours under the basic culture conditions.

B) Experimental results

The MTT assay results are shown in Fig. Statistical analysis of the experimental data was performed by using Microsoft Excel program and post - test was performed using the Bonferroni multiple comparison test to verify the difference between the groups. As shown in FIG. 2, the groups treated with the extracts of overpruising and overprying extracts (hexane, ethyl acetate, butanol and water fractions) were indicated by * as compared with that of cisplatin and 50 μg / ml, 100 μg / Ml and 200 μg / ml, respectively (statistically significant level, p <0.05). It was confirmed that the hexane, butanol and water fractions were increased at 50 μg / ml, 100 μg / ml and 200 μg / ml at 50 μg / ml and 100 μg / ml for the ethyl acetate fraction of the over-phosphorus extract Significance level was p <0.001). In addition, after selecting the concentration through the control value comparison concentration of the selected drugs, the degree of cellular activity of the drug itself (fraction of overproduction and overproduction of the extract) and the amount of drug (fraction of perilla extract and perilla extract) and mixture of cisplatin Cell activity was compared with Control. The results are shown in FIG. 3, and the statistical significance level was p <0.001.

2) Measurement of ROS production

To determine the antioxidant efficacy of the over-extracted extract fraction (hexane, ethyl acetate, butanol and water fractions) of Example 2 above in renal cells, an ROS production experiment was performed.

A) Experimental method

Cells were seeded at 1 × 10 ⁵ cells / well in a 96-well black plate with porcine renal epithelial cells. After culturing for 24 hours, the MTT assay and the crystal violet assay were used to determine the effective fractions of the target drug as dilution factors of the final concentration, followed by dilution and pretreatment. The drug (hexane, ethyl acetate, butanol And water fractions), cisplatin was added to a final concentration of 25 / / ml for 2 hours and cultured for 24 hours. The DCF-DA was dissolved in PBS in the dark room according to the concentration of the DCF-DA and the reaction time determined by the preliminary experiment, and reacted for 5 hours in the basic culture condition. Fluorescence wavelengths were observed at end points of Ex: 285 nm, Em: 528 nm, and S = 50 for ROS generation rates of 8, 12, 16, 20 and 24 hours, respectively.

B) Experimental results

The results of ROS production measurement are shown in FIG.

As shown in FIG. 4, it was confirmed that treatment with the hexane fraction, ethyl acetate fraction, butanol fraction and water fraction, which are fractions of the over-phosphorus extract, inhibited ROS generation by cisplatin. It was found to be similar to antioxidant efficacy in controls not treated with cisplatin.

3) GSH content measurement

GSH levels were examined to determine the cytoprotective effect of the drug on cell damage and to confirm the cytoprotective effect of the over-phosphorus extract and over-phosphorus extract fraction (hexane, ethyl acetate, butanol and water fraction) on cisplatin .

A) Experimental method

The GSH levels were determined by monochlorobimane employing a method and the GSH content measurement experiment was performed with reference to what was performed by Fernandez-Checa and Kaplowitz. Historical data and preliminary experiments were used to determine the number of cells capable of detecting GSH, and the cells were seeded at 2 × 10 ⁶ cells / well in a culture dish. After culturing for 24 h, MTT assay and crystal violet assay were used to determine the drug to be an effective drug considering the dilution factor of the final concentration, followed by dilution and pretreatment. Cisplatin was added to the final concentration of 15 ㎍ / ㎖ And cultured for 24 hours. Cells were harvested using 0.25% trypsin-EDTA and centrifuged at 1200 rpm for 3 min. The cells were centrifuged at 10,000 g for 15 min. The cells were sonicated for 30 sec. To quantify the amount of protein. To the supernatant, 1 g / ml of MPA diluted in DW was added at a ratio of 4: 1 relative to the volume. Vortexing immediately and centrifugation at 10,000 g for 15 minutes were used to collect the deprotonated supernatant. Lt; RTI ID = 0.0 &gt; -20 C. &lt; / RTI &gt;

The porcine renal epithelial cells were cultured in 96 well black plates with 0, 0.2, 0.4, 0.8, 1.2, 1.6, 2.0 μg of GSH Standard (L-Glutathione reduced, Sigma-Aldrich) / Ml, and 20 μl of each sample was dispensed in two or more cycles, and 180 μl of Buffer was added to each well. 10 μl of OPA (Phtthaldialdehyde, Sigma-Aldrich) diluted in methanol at 1 mg / ml was added to each well. After 15 min of reaction at room temperature, the reaction was terminated at Ex: 360 nm and Em: The fluorescence wavelength was measured.

B) Experimental results

Fig. 5 shows the GSH content measurement results for confirming the cytoprotective effect of the perilla extract and the fraction of the perilla extract (hexane, ethyl acetate, butanol and water fraction). Statistical analysis of the experimental data was performed by using Microsoft Excel program and post - test was performed using the Bonferroni multiple comparison test to verify the difference between the groups.

As shown in FIG. 5, the groups treated with the extracts of overprying and over-phosphorus extracts (hexane, ethyl acetate, butanol and water fractions) as compared with cisplatin were shown as *, and the fraction of perilla extract or perilla extract (Hexane, ethyl acetate, butanol, and water fractions) were added to the cells, and it was confirmed that the cells had protective effect (statistically significant level, p <0.05).

Experimental Example  3: in vivo  Acute renal failure efficacy experiment

1) Experimental animal and sample administration

A) Experimental animals

48 male Sprague-Dawely rats (6-week old on receipt, Korea BioLink, Korea) were used for the experiment through 7 days of purification procedure. During the purification and the experiment, the temperature (20-25 ℃) Three animals were housed in a polycarbonate breeding box for the rats in a controlled room (30-35%). The light (dark) cycle was adjusted to 12 hours. The feed (Samyang, Korea) Respectively. All laboratory animals used in this study were treated in accordance with the Guide for the Care and Use of Laboratory Animals [Department of Health, Education, and Welfare Publication (National Institute of Health) 85-23].

B) Sample administration

The perilla extract prepared in Example 1 was used as a sample. Sample administration was divided into five groups: normal control (control) and drug administration group (perilla extract). Samples were administered orally and the dosing period was once a day for a total of 28 days. To induce acute renal failure, cisplatin (5 mg / kg) was injected into the abdomen 5 days before the final sacrifice. Specific sample administration conditions are shown in Table 1 below.

Experimental group Dosage materials and dosage CONTROL INTACT Nothing Vehicle DW + physiological saline (5 ml / kg) Cisplatin DW + Cisplatin (5 mg / kg) EXPERIMENT
(Extract of overproduction) LOW Low dose (100 mg / kg) + Cisplatin (5 mg / kg) MEDIUM Medium dose (300 mg / kg) + Cisplatin (5 mg / kg) HIGH High dose (500 mg / kg) + Cisplatin (5 mg / kg)

2) Measurement of weight change by acute renal failure

The acute renal failure efficacy of the perilla extract prepared in the above Example was confirmed as follows.

A) Experimental method

Regarding the change in body weight, the body weight was measured every 3 days after the fasting for 6 hours. The body weight was measured the day before the autopsy. Weight gain was measured by pre Cisplatin treated periods (after cisplatin pretreatment), after Cisplatin treated periods (after cisplatin injection), and throughout experimental periods (total body weight gain).

B) Experimental results

The results of the weight change measurement after cisplatin treatment in an acute renal failure animal model are shown in Fig. 4 as Mean ± SEM (standard error of mean). As shown in FIG. 4, when the perilla extract was treated at different concentrations, the effect of inhibiting weight loss on cisplatin was shown. In particular, treatment with the extract (100 mg / kg, 500 mg / kg) resulted in (-8.30 ± 3.5 g), (-7.00 ± 0.7 g), about 73% of the induced cisplatin (-11.30 ± 7.10 g) %.

3) BUN (Blood Urea Nitrogen) measurement

The acute renal failure efficacy of the perilla extract prepared in the above Example was confirmed by measuring the amount of BUN (Blood Urea Nitrogen) activation as described below.

A) Experimental method

All animals were fasted for more than 18 hours on the day before and after the cisplatin injection. Approximately 0.5 ml of blood was collected from the orbital vein and left at room temperature for 1 hour. The serum was separated by centrifugation at 3,000 rpm. After that, blood BUN content was measured using an automatic blood analyzer (Toshiba 200 FR, Japan). In order to minimize the difference in blood BUN content due to individual differences before cisplatin injection, the changes in blood BUN of final sacrifice days before cisplatin injection were measured using the following formula.

* Changes (After Cisplatin injection) = Serum BUN levels at sacrifice - Serum BUN levels at 23 days after test article treatment (at Cisplatin injection)

B) Experimental results

The results of BUN measurement after cisplatin treatment in an acute renal failure animal model are shown in Fig. 5 as Mean ± SEM (standard error of mean). Statistical analysis of the experimental data was performed by using Microsoft Excel program and post - test was performed using the Bonferroni multiple comparison test to verify the difference between the groups. Statistical significance was p <0.05, and compared to cisplatin, the group treated with the extract was treated with *. As shown in FIG. 5, when the cisplatin treatment was carried out, the BUN level was increased by about 3 times or more at 83.0 ± 12.7 mg / dl, and the increase in the BUN level was inhibited in the extract- kg, 300 mg / kg, 500 mg / kg), the inhibitory effect on the BUN increase was found to be 66% or lower in the cisplatin-induced group.

4) Blood creatinine measurement

The acute renal failure efficacy of the perilla extract prepared in the above examples was confirmed by measuring creatinine amount as follows.

A) Experimental method

All the animals were fasted for more than 18 hours on the day immediately before and after the cisplatin injection. Approximately 0.5 ml of blood was collected from the orbital vein and left for 1 hour at room temperature. Then, the serum was separated by centrifugation at 3,000 rpm. After that, blood creatinine content was measured by an automatic blood analyzer. In order to minimize the differences in blood creatinine levels due to individual differences before cisplatin injection, the changes in blood creatinine at final sacrifice days before cisplatin injection were measured using the following formula.

* Changes (After Cisplatin injection) = Serum Creatinine levels at sacrifice - Serum Creatinine levels at 23 days after test article treatment (at Cisplatin injection)

B) Experimental results

The creatinine measurement results after cisplatin treatment in an acute renal failure animal model are shown in Fig. 6 as Mean ± SE (standard error of mean). Statistical analysis of the experimental data was performed by using Microsoft Excel program and post - test was performed using the Bonferroni multiple comparison test to verify the difference between the groups. Statistical significance was p <0.05, and compared to cisplatin, the group treated with the extract was treated with *. As shown in FIG. 6, the creatinine level was increased to about 3.times., Which was about 3.times. 1.7 mg / dl, when cisplatin was applied. Creatine levels were 100 mg / kg, 300 mg / mg / kg of cisplatin-induced creatinine.

Claims (12)

A pharmaceutical composition for the prevention or treatment of acute renal failure containing an over-the-whole extract or a fraction of an over-extracted extract as an active ingredient.
2. The method of claim 1, wherein the extract is a gwaru gwaru of water, the pharmaceutical composition being prepared by extracting with C _1-4 alcohol or mixtures thereof.
The pharmaceutical composition according to claim 2, wherein the extraction is an ultrasonic extraction, a room temperature extraction, a hot water extraction, a cold extraction, a reflux cooling extraction, or a steam extraction.
The pharmaceutical composition according to claim 1, wherein the fraction of the over-phosphorus extract is a hexane fraction, an ethyl acetate fraction, a butanol fraction or a water fraction.
The pharmaceutical composition according to claim 1, wherein the acute renal failure is caused by an anticancer agent.
The pharmaceutical composition according to claim 5, wherein the anticancer agent is cisplatin.
A functional food composition for preventing or ameliorating acute renal failure containing an extract of perilla or extract of perilla as an active ingredient.
The method of claim 7, wherein the extract is a gwaru gwaru of water, C _{1 -4} alcohol or a functional food composition, characterized in that is made by extraction with a mixed solvent thereof.
The health functional food composition according to claim 8, wherein the extraction is ultrasonic extraction, room temperature extraction, hot water extraction, cold extraction, reflux cooling extraction or steam extraction.
The health functional food composition according to claim 7, wherein the fraction of the over-extracted extract is hexane fraction, ethyl acetate fraction, butanol fraction or water fraction.
The health functional food composition according to claim 7, wherein the acute renal failure is induced by an anticancer agent.
12. The health functional food composition according to claim 11, wherein the anticancer agent is cisplatin.

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