WO2011034362A2 - Composition containing saururus chinensis extract or sauchinone as an active ingredient, for preventing and treating diseases caused by the overexpression of lxr-α - Google Patents

Abstract

The present invention relates to a composition containing Saururus chinensis extract or Sauchinone as an active ingredient, which inhibits the expression or activity of liver X receptor α (LXRα) or sterol response element binding protein-1 (SREBP-1), and which can thus be used in preventing, improving, and treating diseases caused by the overexpression or activity of LXRα or SREBP-1. The composition of the present invention exhibits remarkable effects of inhibiting adipogenic gene expression, and further inhibiting the accumulation of triglycerides in liver tissue, and therefore is advantageous in treating fatty liver caused by metabolic disorders resulting from a high-fat diet. In addition, the composition of the present invention can be effectively used in preventing, improving, and treating, for example, liver diseases such as fatty liver caused by the accumulation of liver lipids, hypertriglyceridemia, hyperreninemia, renin-induced hypertension, aldosteronism, adrenoleukodystrophy, glomerulosclerosis, proteinuria, renal failure, etc.

Description

Composition for the prevention and treatment of diseases caused by overexpression of L-X-alpha containing three hundred sec extract or sacchinone as an active ingredient

The present invention relates to the pharmaceutical or food use of the three hundred sec extract or Sauchinone. More specifically, diseases caused by excessive activity of LXRα (Liver X receptor-alpha) or SREBP-1 (Sterol Response Element Binding Protein-1) using Trichophyllum extract or Succinone, for example, due to accumulation of liver lipids such as fatty liver It is associated with the prevention, improvement and treatment of liver disease, hypertriglyceridemia, hyperreninemia, high blood pressure caused by renin, aldosteroneism, adrenal protein dystrophy, renal glomerulosis, proteinuria, nephropathy.

The present invention is derived from a study performed as part of the Ministry of Education, Science and Technology (ERC) project [Task No .: R11-2007-107-01001-0, Title: New Drug Development Center for Metabolic and Inflammatory Diseases]

Sambaekcho (三 白草) is the terrestrial part of the three hundred perennial perennial plant (Saururus chinensis Baill.). Sambaekcho is a long-lived grass that rarely grows in the wet soil of southern Korea, including Jeju Island. It is 30 ~ 90cm tall and its root is white and hairy. Stems grow straight and the leaves are egg-shaped, and in early summer, two or three leaves under the stem tip flower turn white.

Three hundred seconds is lignan, methyl-n-nonyl-ketone, tannin, guercitrin, isoquercitrin, abicurin ( It contains ingredients such as avicularin) and is known to be effective in treating gynecological diseases such as antioxidant, anti-aging, anticancer (liver cancer, lung cancer, stomach cancer), cold sores, and menstrual disorders. In addition, the ingredients such as querchitrin, isoquerchitrin, and pravonoids contained in three hundred seconds is used to remove stool because it has a good effect on bowel movement.

Sauchinone is a lignan-based substance that is one of the components of 300 seconds and is known to have cytoprotective, anti-inflammatory and antioxidant effects.

Liver X receptor (LXR) is a type of nuclear hormone receptor, a gene involved in cholesterol metabolism and homeostasis, such as apolipoprotein E (apoE), ABCA1, ABCG1, ABCG5, ABCG8, cholesterol 7α-hydrate Plays an important role in the transcriptional regulation of cholesterol 7α-hydroxylase and scavenger receptor class B type I genes [Schwarz et al., Biochem. Biophys. Res. Commun., 2000, 274: 794-802. LXR also acts directly on the SREBP-1c gene to regulate lipid metabolism [Yoshikawa et al., Mol. Cell. Biol., 2001, 21: 2991-3000.

LXR isomers include LXRα and LXRβ. LXRα is mainly present in the liver and LXRβ is expressed mostly in organs. LXRα is activated by natural ligands, oxysterols, high concentrations of sugars and artificial ligands, T0901017, GW3965 and the like, and regulates the expression of genes responsible for lipid formation and cholesterol homeostasis in the body. LXRα acts as a lipid sensor in the production of liver lipids, which strongly increases the expression and activity of SREBP-1c, a key transcription factor that regulates the expression of lipid-forming genes, thereby promoting fatty acid synthesis in liver tissues and increasing blood triglyceride levels. .

There are largely SREBP-1c-dependent pathways and SREBP-1c-independent pathways in which LXRα activation causes nonalcoholic fatty liver. In SREBP-1c-dependent fatty liver, the expression of lipogenetic genes is upregulated through transcriptional activation of LXRα-mediated SREBP-1c, and in SREBP-1c-independent fatty liver, LXRα activation increases the expression of CD36 protein, a transporter of free fatty acids. By stimulating the transport of fatty acids to the liver.

It has also been reported that LXRα plays an important role in the secretion of renin in the kidney. LXRα and LXRβ are both expressed in abundance in juxtaglomerular cells that produce renin. According to Morello et al., T0901017 or GW3965, an agonist of LXRα, increases the expression of renin mRNA in the kidney and increases blood renin activity [Mello et al., J. Clin. Invest., 2005, 115: 1913-1922. Excessive increase in blood renin leads to hyperreninemia, which leads to hypertension and aldosteroneism.

LXR also regulates the expression of the ABCD2 gene, which is associated with adrenoleukodystrophy (ALD), a rare disease in which the body's `` very long chain fatty acid '' (VLCFA) does not break down and enters the brain and destroys nerve cells. It has been reported to be useful for the treatment of ALD [Winehopper et al., J. Biol. Chem., 2005, 280: 41243-41251.

SREBP (Sterol Response Element Binding Protein) is a protein that binds to sterol response element (SRE), a transcriptional regulatory region of sterol-regulated genes, and exists in three isoforms. SREBP-1a and SREBP-1c are transcribed from the same gene and SREBP-2 is expressed from other genes. SREBP-1c is a gene mainly involved in fatty acid synthesis, and SREBP-2 is a transcription factor that regulates transcription of genes involved in cholesterol synthesis.

SREBP-1 and SREBP-2 increase in the expression of kidney with age and thus increase the lipid synthesis and accumulation of triglyceride and cholesterol in the kidney, which are called glomerulosclerosis, proteinuria, kidney It has been reported to play an important role in the nephropathy [Jiang et al., Kidney Int., 2005, 68 (6): 2608-2620].

SREBP is usually present as a endoplasmic reticulum membrane protein and is 125 kilodaltons (kDa) in size. Before being activated by stimulation such as sterol depletion, it is bound to the membrane in an inactivated form, and upon activation, it is transferred to the Golgi body and cleaved into an active protein of 65 kilodalton size. When activated, SREBPs move into the nucleus and bind to the SRE of the target gene, thereby increasing the expression of liposynthetic genes. Target genes of SREBP-1c include fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and stearoyl CoA desaturase (SCD). If the amount of free fatty acids and fatty acids directly produced by the liver from the blood is higher than the amount of VLDL or beta-oxidized fatty acids, the lipid metabolism balance in the liver is broken down and is transferred to the fatty liver. , SREBP-1c, which regulates FAS, ACC, SCD, etc., which are responsible for lipid synthesis in the liver, are important factors for alcoholic or non-alcoholic fatty liver [Kohjima et al., Int. J. Mol. Med., 2008, 21 (4): 507-511, Donohue, World J. Gastroenterol. 2007, 13 (37): 4974-4978.

Fatty liver is a medical condition in which fat exceeds more than 5% of the total liver weight. Liver disease, including this, is known to be the second most serious disease after cancer in developed countries in the 40-50 adult population. About 30% of the population in major countries, including developed countries, already has fatty liver symptoms, and 20% of them progress to cirrhosis through liver fibrosis. Half of these cirrhosis patients die of liver disease within 10 years of diagnosis.

Alcoholic fatty liver disease is caused by the accumulation of fatty acids in the liver by metabolites that occur when alcohol is metabolized to acetaldehyde through acetate. Non-alcoholic fatty liver disease is associated with a number of factors (obesity, type 2 diabetes, hypertension, hyperlipidemia, metabolic syndrome, etc.) or lipid metabolism abnormalities in normal individuals with or without significantly lower alcohol consumption. It occurs through stress and abnormal cytokine production. This is a broad concept that includes everything from simple fatty accumulation to fatty tissue inflammation (such as hepatitis) and some steatohepatitis, and fatty liver from these fat accumulations is known as de novo lipogenesis. Genes associated with inflammatory pathways can be activated to cause fatty hepatitis.

Currently, there are few drugs useful for treating fatty liver pharmacologically, and only exercise and diet are recommended. However, the treatment efficiency of fatty liver by these methods is very low, and there is an urgent need for the development of effective therapeutic agents. Betaine, glucuronate, methionine, choline, and lipotrophic agents may be used as adjuvant, but the pharmacological evidence for them is fully demonstrated. It is not. Therefore, it is urgent to develop a safe fatty liver treatment that is excellent in effect but does not cause side effects. In particular, it is necessary to develop a substance capable of effectively controlling lipid metabolism in consideration of the aforementioned mechanism of fatty liver development.

In the present invention, it is an object of the present invention to find and provide an inhibitor of LXRα or SREBP-1. Diseases caused by overexpression or overactivity of LXRα or SREBP-1, such as liver disease due to accumulation of liver lipids such as fatty liver, hypertriglyceridemia, hyperreninemia, hypertension due to renin, aldosteroneism, adrenal protein dystrophy, gentleman It is also a problem to be solved by the present invention to provide a pharmaceutical or food composition for the prevention, improvement and treatment of concrete hardening, proteinuria, kidney damage and the like.

In order to solve the above problems, the present inventors have provided a pharmaceutical composition or food composition useful for the prevention, improvement and treatment of diseases caused by the overexpression or overactivity of LXRα or SREBP-1 comprising three hundred s. Extract or sacchinone as an active ingredient. to provide.

 The composition containing the tritical herb extract or sacchinone of the present invention as an active ingredient can be effectively used for the prevention, improvement and treatment of diseases due to overexpression or overactivity of LXRα or SREBP-1. Administration of the composition of the present invention inhibits the expression and activity of SREBP-1, a key transcription factor that regulates lipid production enzyme gene expression through the regulation of LXRα activation and further inhibits the expression of lipid production genes. The composition of the present invention, which inhibits the accumulation of triglycerides in liver tissue and contains them as an active ingredient, can be usefully used for preventing and treating liver diseases caused by accumulation of liver lipids such as fatty liver. In addition, the composition is hypertriglyceridemia, hyperrenemia, hypertension due to renin, aldosteroneism, adrenoleukodystrophy, glomerulosclerosis, proteinuria, nephropathy, and the like. It can be effectively used for prevention, improvement and treatment.

The composition of the present invention can be used for the manufacture of medicines or nutraceuticals that can be used for the prevention, amelioration or treatment of the disease.

1 shows the results of observing the effect of sacchinone on the expression level of LXRα by treating T0901317, an LXRα activator, with HepG2, a hepatocyte cell line. (Con: no treatment group, **: p <0.01 compared with Con group, ##: p <0.01 compared with T0901317)

2 shows the effect of T0901317, an LXRα activator, on hepG2, a hepatocyte cell line, and the effect of Succinone treatment on LXRE. (Con: untreated group, **: p <0.01, ## compared to Con group) : P <0.01) compared to the group treated with T0901317 only

Fig. 3 shows the results of treatment of TX01α317, an LXRα activator, with hepG2, a hepatocyte cell line, and the effect of sacchinone on the expression level of SREBP-1 protein. (Con: untreated group, **: p compared with Con group) <0.01, #: p <0.05 compared to the group treated only with T0901317, ##: p <0.01 compared with the group treated only with T0901317)

Figure 4 shows the results of confirming the effect of the Succinone treatment on the weight change in the Succinone administration schedule and a high-fat diet-induced fatty liver model. (ND: Dietary HFD: High-fat diet Sau: Sauchinone, **: p <0.01 compared to ND group: p <0.05 compared to HFD alone group)

5 is a result of observing the effect of Succinone treatment on the hepatic expression level of lipid-generating genes (LXR, SREBP) induced by a high lipid diet. RXRα or SREBP-1 mRNA levels in the normal diet were expressed as relative values. (ND: normal diet HFD: high-fat diet Sau: Sauchinone **: p <0.01 compared to ND group p <0.01 compared to HFD alone group)

6 is a result of observing the effect of the administration of the sacchinon on the content of triglycerides in the liver tissue in a fatty liver animal model induced by a high-lipid diet. (ND: normal diet HFD: high-fat diet Sau: Sauchinone **: p <0.01 compared to ND group ##: p <0.01 compared to HFD alone group)

FIG. 7 is a micrograph of a fatty liver animal model induced by a high-lipid diet and liver tissue stained by fat stain method (Oil Red O) when the sacchinone was administered to the animal model (x100).

8 is a result confirming the effect on the ALT activity of hepatocytes when administration of sacchinone in a fatty liver animal model induced by a high-fat diet. (ND: normal diet HFD: high-fat diet Sau: Sauchinone **: ND group P <0.01 compared with #: p <0.01 compared to HFD alone)

Figure 9 is a micrograph of the results of H & E staining of liver tissue when administration of sacchinone in a high-fat diet-induced fatty liver animal model. (Scale bar = 200um (low magnification, left column) or 50um (high magnification, middle and right) Column) arrow: hepatocellular necrosis, arrow head: infiltration of inflammatory cells, asterisk: fat accumulation in liver tissue, P: contextual space HFD: high-fat diet Sau (Low): Sauchinone 10mg / kg Sau (High): Sauch Paddy field 30mg / kg)

FIG. 10 shows the results of treatment of HepG2, a hepatocyte cell line, T0901317, which is an LXRα activator, and the effect of treatment with 300 sec extract for LXRE. (CONTROL: no treatment group, **: p <0.01, ##: P <0.01) compared to the T0901317 only group

11 is a result confirming the effect of trichophytium extract treatment on body weight change in a high-fat diet-induced fatty liver animal model. (ND: normal diet HFD: high-fat diet SE: trichophytium extract Sau: Sauchinone, **: P <0.01 #: compared to ND group p <0.05 ##: p <0.01 compared to HFD alone group)

12 is a result of observing the effect on the content of triglycerides in liver tissue when administration of trichophytium extract in a fatty liver animal model induced by a high-fat diet. (ND: dietary diet HFD: dietary lipid SE: triglyceride extract Sau: Saucinone **: p <0.01 compared to ND group ##: p <0.01 compared to HFD alone group)

13 is a result showing the oil red O index (Oil Red O index) when staining the liver tissue in the fat liver animal model induced by a high lipid diet and the three hundred seconds extract in the animal model (ND: Normal diet HFD: High-fat diet SE: Three hundred sec extract Sau: Sauchinone **: p <0.01 ## compared to ND group p <0.01 compared to HFD alone group).

FIG. 14 is a micrograph of the staining result of FIG. 13 (x100). FIG.

15 is a result confirming the effect on the ALT activity of hepatocytes in administration of trichophytium extract in a fatty liver animal model induced by a high-fat diet. (ND: normal diet HFD: high-fat diet SE: trichophytium extract Sau: Sauchinone * *: P <0.01 # compared to ND group: p <0.05 ## compared to HFD alone group p <0.01 compared to HFD alone group)

Figure 16 is a micrograph showing the results of H & E staining of liver tissue when administration of trichophytium extract or sautinone in a high-fat diet-induced fatty liver animal model (arrow: hepatocyte necrosis, arrow head: infiltration of inflammatory cells). , Asterisk: fat accumulation in liver tissue, C: central vein)

The present invention inhibits the expression or activity of LXRα (Liver X recptorα) or SREBP-1 (Sterol Response Element Binding Protein-1) comprising three hundred sec extract or Sauchinone represented by Formula 1 as an active ingredient. Thereby providing a composition for preventing, ameliorating and treating diseases due to their overexpression or activity.

Formula 1

The pharmaceutical composition or the food composition of the present invention is a liver disease caused by accumulation of liver lipids, such as fatty liver, hypertriglyceridemia, hyperreninemia, high blood pressure caused by renin, aldosteroneism, adrenal protein dystrophy, renal glomerulosis, proteinuria, kidney damage, etc. It can be effectively used for prevention, improvement and treatment. The composition of the present invention exhibits a remarkable effect on the inhibition of the expression of lipid-generating genes, and further on the accumulation of triglycerides in liver tissue, and is effective for fatty liver due to metabolic disorders induced by a high lipid diet.

Hereinafter, the present invention will be described in detail.

The three hundred seconds extract of the present invention can be prepared to include a large amount of sacchinone by extracting three hundred seconds with a low polar solvent, alcohol or a mixture thereof. In this case, any low polar solvent may be used as long as it is commonly used in the art, such as chloroform, dichloromethane, ether, or ethyl acetate. Alcohols include those commonly used as extraction solvents in the art, such as methanol, ethanol, propanol, n-propanol, isopropanol, butanol, n-butanol, and lower alcohols are preferred. The manufacturing process of the three hundred seconds extract of the present invention may further include the step of first extracting with a non-polar solvent to remove the non-polar components before the extraction process. The non-polar solvent used at this time may include n-hexane, cyclohexane, benzene or petroleum ether, but is not limited thereto.

In one embodiment of the present invention, n-hexane, cyclohexane, benzene or petroleum ether is added to the shade of three hundred seconds in dry shade for 1 to 7 days, preferably 24 to 72 hours, extracted at 10 to 100 ℃ n After the extract containing the hexane soluble component was removed, a low polar solvent such as chloroform, dichloromethane, ether or ethyl acetate, alcohol or a mixture thereof was added thereto for 1 to 7 days, preferably 24 to 72 hours. Extraction method such as reflux cooling extraction, cold extraction method, ultrasonic extraction method, hot needle extraction method, supercritical extraction method, etc., preferably 100 seconds to extract 100 seconds by using ultrasonic extraction method Can be prepared.

The extract includes an extract obtained by extraction, a diluent or concentrate of the extract, a dried product obtained by drying the extract or a crude or purified product of the extract.

The present invention provides a method for preparing the extract of three hundred seconds described above.

In another aspect, the present invention provides a pharmaceutical or food composition and a pharmaceutical, health functional food prepared therefrom comprising three hundred seconds extract containing a large amount of saucchinone obtained through the manufacturing process as an active ingredient.

 Succinone used in the present invention can be obtained by separating the three hundred sec extract of the present invention by chromatography, and further recrystallization and the like can be carried out to obtain more purified sautinone.

The sautinone may be represented by Formula 1 and includes all possible isomers from the above formula. Specifically, Formula 1 has six chiral centers, and the sautinone of the present invention may be any one of the 64 optical isomers or a mixture of one or more of the isomers.

 The inventors of the present invention have surprisingly found that the expression and activity of LXRα and SREBP-1 that is increased when T0901317, known as an activator of LXRα, is treated by hepatocytes, is inhibited by sacchinone or trichophytium extract comprising the same. (Figs. 1-3 and 10). In vivo experiments also revealed that sacchinone also inhibited the expression of LXRα and SREBP-1 in mice fed with a high lipid diet (FIG. 5).

In addition, when administration of a high fat diet to increase the content of triglycerides in the liver tissue, the administration of the sacchinone or three hundred seconds extract, significant weight loss effect (Figs. 4 and 11), the inhibitory effect of the increase in triglycerides 6 and 12, the clinical efficacy was confirmed by observing a significant decrease in the amount of fat in the liver tissue by lipo staining method (Figs. 7 and 13-14). In mice treated as described above, the elevation of ALT was significantly suppressed (FIGS. 8 and 15), and necrosis and inflammation of tissues caused by liver fat accumulation were reduced (FIGS. 9 and 16). Inhibitory effect against fatty liver and hepatitis was confirmed.

These results demonstrate that the three hundred sec extract or sacchinone according to the present invention directly contributes to the reduction of cholesterol and triglycerides by regulating gene expression and protein synthesis involved in lipid metabolism in the liver. Therefore, in contrast to conventional hepatoprotective agents or therapeutic agents for the treatment of liver diseases, by controlling the expression of enzymes involved in antioxidant or detoxification, the method of activating the defense mechanism, the present invention is directed to fatty liver by directly controlling the accumulation of lipids in the liver. Immediate and immediate therapeutic effects can be expected.

On the basis of this, the present invention inhibits the expression and activity of LXRα (Liver X recptorα) and SREBP-1 (Sterol Response Element Binding Protein-1) comprising three hundred sec extract or sacchinone as an active ingredient, thereby overexpressing them or It provides a pharmaceutical composition or food composition for the prevention, improvement and treatment of diseases due to activity. In addition, the present invention provides a method of inhibiting the expression or activity of LXRα or SREBP-1 using a tritical herb extract or sacchinone, and a method for preventing, ameliorating or treating a disease due to their overexpression or activity.

Trichophytium extract or Succinone inhibits the expression and activity of SREBP-1, a key transcription factor that regulates the expression of lipid-forming enzyme genes through the regulation of XRα activation, and furthermore, hepatic tissues caused by metabolic disorders through inhibition of the expression of lipid-forming genes. Since the accumulation of triglycerides is suppressed, the composition of the present invention containing the same as an active ingredient can be usefully used for preventing and treating liver disease caused by accumulation of liver lipids such as fatty liver. The fatty liver may be alcoholic fatty liver or non-alcoholic fatty liver, and non-alcoholic fatty liver includes nutrient fatty liver, hunger fatty liver, obese fatty liver and diabetic fatty liver. Liver diseases caused by the accumulation of hepatic lipids include fatty liver disease, liver fibrosis, cirrhosis of the liver, hypertrophy of the liver, liver failure, liver failure, gallstones and portal hypertension.

In addition, the composition of the present invention is a disease caused by hyperactivity of LXRα or overexpression and activity of SREBP-1, hypertriglyceridemia, hyperreninemia, high blood pressure caused by renin, aldosteroneism, adrenal protein dystrophy, renal glomeruli, proteinuria, kidney It may be used for the prevention, improvement or treatment of harm, etc., but is not limited thereto.

The triticale extract or sautinone included in the composition of the present invention may be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art. Pharmaceutically acceptable salts of the three hundred sec extract or sautinone above include salts of acidic or basic groups which may be present in the compound, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through. As the pharmaceutically acceptable salts of the present application, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equivalent molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like.

 Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salts as metal salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

 Accordingly, the present invention includes one or more selected from the group consisting of sacchinone, trichoola extract, pharmaceutically acceptable salts thereof, and solvates or hydrates thereof, and includes a pharmaceutically acceptable carrier. It provides a pharmaceutical composition for the prevention and treatment of liver disease, hypertriglyceridemia, hyperreninemia, renin-induced hypertension, aldosteroneism, adrenal protein dystrophy, renal glomerulosis, proteinuria, and kidney damage caused by the accumulation of liver lipids such as fatty liver.

 The composition of the present invention preferably contains 0.01 to 99.9% of sacchinone or triticale extract, and more preferably 0.1 to 90%. However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

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

 The composition comprising the tritical herb extract or sacchinone according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterilization, respectively, according to a conventional method. Carriers, excipients and diluents which may be formulated in the form of injectable solutions and may be included in the compositions 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 include at least one excipient such as starch, calcium carbonate, or the trichoola extract or sautinone. , Sucrose or lactose, gelatin and the like are mixed and prepared. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Oral liquid preparations include suspending agents, liquid solutions, 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. Witsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like may be used as the base material of the suppository.

 Preferred dosages of the compositions of the present invention vary depending 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, the composition containing the three hundred ss extract or sacchinone of the present invention as an active ingredient is administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day Good to do. 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.

 The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, endometrial, intraperitoneal or intracerebroventricular injection.

 In another aspect, the present invention contains three hundred seconds extract or sachinone as an active ingredient. Provides health functional foods for the prevention and improvement of liver disease, hypertriglyceridemia, hyperreninemia, renin-induced hypertension, aldosteroneism, adrenal protein dystrophy, renal glomerulus, proteinuria and kidney damage .

 The composition containing the tritical herb extract or sacchinone of the present invention can be used in a variety of drugs, foods and beverages effective for the prevention and improvement of the disease. Foods to which the composition of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

 Since the extract of three hundred seconds or sachinone of the present invention has little toxicity and side effects, it is a drug that can be used with confidence even when taken for a long time for the purpose of prevention and improvement.

 The triticale extract or sautinone of the present invention is a liver disease caused by accumulation of liver lipids such as fatty liver due to metabolic disorders, hypertriglyceridemia, hyperreninemia, hypertension caused by renin, aldosteroneism, adrenal protein dystrophy, renal glomerulosclerosis, proteinuria And food or beverage for the purpose of preventing and improving kidney damage. In this case, the amount of the compound in the food or beverage may generally be added in an amount of 0.01 to 15% by weight of the total food weight in the case of the health food composition, and 0.02 to 10 g, preferably 100 ml in the case of the health beverage composition. Can be added in a ratio of 0.3 to 1 g.

 In addition to containing the compound as an essential ingredient in the indicated ratio, the health beverage composition of the present invention is not particularly limited in the liquid component, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates are, for example, monosaccharides such as glucose and fructose, for example disaccharides such as maltose and sucrose, and polysaccharides such as dextrin, cyclodextrin, and the like. Sugars 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, 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 and 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 from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

All technical terms used in the present invention are used in the meaning as commonly understood by those skilled in the art in the related field of the present invention, unless otherwise defined. Also described herein are preferred methods or samples, but similar or equivalent ones are included in the scope of the present invention. The contents of all publications described herein by reference are incorporated into the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following Examples and Experimental Examples are only illustrative of the present invention in detail, and the scope of the present invention in any sense is not limited to the following.

Reference Example 1: Experimental Animals and Diet

Male C57BL / 6 mice (average weight 25-30 g) used as experimental animals were purchased from Charles River Orient (Seoul, Korea). Before the experiment, animals were adapted from the Seoul National University College of Pharmacy Animal Experimental Research Building with controlled humidity of 55 ± 5%, temperature of 22 ± 2 ℃ and ventilation for more than 1 week. Changed the contrast in time periods. No significant changes were observed in the amount of food and water for the duration of the experiment. Animal weights and conditions were examined once weekly and were bred for 11 weeks (8 weeks for the Triticale extract experiment) with a high fat diet (HFD) (60% fat) (Dyets Inc., Bethlehem) or normal diet. Succinone (10 or 30 mg / kg, 5 times / week) or 300 seconds extract (10, 30 or 100 mg / kg, 5 times / week) was administered during the last 5 weeks (4 weeks for the 300 seconds extract experiment). Each group consisted of a total of 10 mice.

Reference Example 2: Sample Preparation

A high-fat diet for fatty liver induction was purchased from Diet Co., USA. The desired concentration of sautinone was prepared by diluting the sautinone prepared in Example 2 with 40% PEG # 400 [PEG # 400: water = 4: 6].

Reference Example 3: Real time-RT PCR

CDNA was obtained using total RNA (2 μg), d (T) ₁₆ primer and AMV reverse transcriptase extracted from mouse liver. Relative amounts of genes were quantified by Realtime RT-PCR method using CyBr green dye. Realtime RT-PCR used Light-cycler ^2.0 from Roche (Mannheim, Germany). PCR was performed according to the manufacturer's method and the relative amount of each gene was analyzed using the Light-cycler software 4.0 program.

Reference Example 4: Western Blot

Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed using a Mighty Small II SE 250 apparatus according to the Laemmli UK method (1970). Dissolve the fraction of liver sample in sample dilution buffer [63mM Tris (pH.6.8), 10% glycerol, 2% SDS, 0.0013% bromophenol blue, 5% β-mercaptoethanol] and use 7.5%, 9% gel Electrophoresis was performed in buffer (including Tris 15g, glycine 72g, SDS 5g in 1L solution). The electrophoresis gel was transferred to nitrocellulose paper for 1 hour at 190 mAmps in transition buffer [25 mM Tris, 192 mM glycine, 20% v / v methanol (pH.8.3)] using a transfer electrophoresis device. After reacting Anti-SREBP-1 as a primary antibody, Horseradish peroxidase-conjugated goat anti-rabbit IgG was reacted with secondary antibody for 1 hour and developed using ECL chemiluminecence system (Amersham, Gaithesberg, MA). The homogeneity of the protein content in the samples was confirmed using anti-β-actin antibodies (Sigma, St. Louis, MO).

Reference Example 5: Analysis Method

The data presented in the experiments were analyzed using a pharmacological calculation program. After testing the significance of various experimental groups by one-way square deviation analysis (Fisher, RA, Statistical Methods for Research Workers , Edinburgh : Oliver & Boyd , 1925) Newman-Kells test (Norman GR et al., Biostatistics: The Bare Essentials, 2000) was determined ( ^{*, #} p <0.05, ^{**, ##} p <0.01).

Example 1. Preparation of three hundred vine extract

Preparation of 300 Seconds Chloroform Extract

15 liters of 100% n-hexane was added to 1.2 kg (Daewon Pharmaceutical Co., Ltd., Daegu) in the shade for 48 hours, and the extract was discarded. Then, 15 liters of 100% chloroform was added, extracted at room temperature for 72 hours, and filtered with a filter paper. The filtered filtrate was concentrated under reduced pressure with a vacuum concentrator (N-21NS, ELYA) to obtain 80 g (SBC-1) of the extract of brown trichophyte in the form of a brown gel. HPLC measurement using a Saucchinone standard confirmed that the extract of Trichophytium contained 5-10% of Sauchinone.

Preparation of 300 Seconds Ethanol Extract

15 liters of 100% n-hexane was added to 1.2 kg of dried finely sliced vinegar for 48 hours, and the extract was discarded. 15 liters of 100% ethanol was added thereto, extracted at room temperature for 72 hours, filtered through a filter paper, and the filtrate was filtered. Concentrated under reduced pressure with a vacuum concentrator (N-21NS, ELYA) to give 60 g (SBC-2) of the extract of brown gel in the form of a brown gel. It was found that% of sacchinone was included.

Example 2. Preparation of Sauchinone

The 300 sec chloroform extract (SBC-1) obtained in Example 1 was loaded on silica gel column chromatography (70-230 mesh, 700 g), and 2 L of n-hexane was poured therein, followed by n-hexane- Ethyl acetate (n-hexane-EtOAc [50: 1 (3L), 30: 1 (5L), 15: 1 (5L)]) was separated by decantation. The n-hexane-ethyl acetate 30: 1 (2L) fraction was concentrated under reduced pressure to obtain 13 g of a light brown small fraction. This fraction was again extracted with n-hexane-ethyl acetate (50: 1 (2 L), 25: 1 (3 L), 9: 1 (2 L), then 1: 1 (1 L)]. Using a gradient solvent gradient, secondary silica gel column chromatography (230-400 mesh, 200 g) was carried out and divided into small fractions again. Fractions 18-25 of the small fractions separated from the secondary silica gel column chromatography were identified as fractions rich in sacchinone [Rf = 0.8, CHCl ₃ -methanol (9: 1)] and concentrated to 5 g. A light brown subfraction was obtained which was more abundant with sacchinone. The small fraction was recrystallized in ether to obtain 2.5 g of pure sautinone.

I. Effect of Sauuccinon

Experimental Example 1 Inhibitory Effect of LXRα Treatment by Succinone Treatment

As a cell line, HepG2, a representative human liver cell line, was used. HepG2 was pretreated with sacchinone (Sau, 30 μM) for 1 hour, treated with LXRα activator T0901317 (10 μM) for 12 hours, mRNA was isolated from hepatocytes, and cDNA was synthesized via RT-PCR. The primers (Human LXR, 5'-GATCGAGGTGATGCTTCTGGAG-3 '(sense) and 5'-CCCTGCTTTGGCAAAGTCTTC-3' (antisense)) were analyzed by real-time PCR.

When the LXRα activator T0901317 was treated with HepG2, a hepatocyte cell line, the mRNA expression of LXRα was significantly increased to 2.67 when the control amount was 1, and was statistically significantly decreased to 0.73 by treatment with sacchinone. (FIG. 1).

Experimental Example 2: LXRα activity inhibitory (LXRE binding capacity inhibition) effect by treatment with Succinone

LXRα regulates gene expression by binding to a specific region (LXRE) present in the target gene promoter. In order to observe whether the LXRE binding capacity was changed by sacchinone treatment, a reporter gene assay (Luciferase reporter gene assay) was performed.

First, a vector containing LXRE was transfected to a HepG2 cell line using lipofectamine (Invitrogen, USA). After 12 hours, HepG2 was pretreated with sacchinone (Sau, 30 μM) for 1 hour and treated with LXRα activator T0901317 (10 μM) for 12 hours. Subsequently, a fraction of the cells was obtained using a passive lysis buffer (Promega, USA), and then luciferase activity was measured using a luminometer.

Luciferase activity was increased to 2.11 when the control activity was set to 1 by TO901317 treatment, and significantly reduced to 1.61 when pretreated with sacchinone (FIG. 2).

Experimental Example 3: Effect of SREBP-1 Expression and Inhibition of Activity by Succinone Treatment

When the hepatocyte cell line HepG2 was treated with T0901317, an LXRα activator, and T091317 and sacchinone, the expression levels of mRNA and protein of SREBP-1 were compared (FIG. 3).

HepG2 was pretreated with sacchinone (Sau, 30 μM) for 1 hour and treated with LXRα activator T0901317 (10 μM) for 12 hours. Thereafter, the proteins were separated and subjected to western blot to observe protein changes in SREBP-1 (FIG. 3, bottom). In addition, mRNA was isolated to synthesize cDNA through RT-PCR, using real primers (human SREBP-1, 5'-CGACATCGAAGACATGCTTCAG-3 '(sense) and 5'-GGAAGGCTTCAAGAGAGGAGC-3' (antisense)) Analysis by time PCR (FIG. 3, top).

When LXRα activator (T0901317) was treated in hepG2, a hepatocyte cell line, mRNA of SREBP-1, a cellular protein that regulates lipogenetic gene expression (Fig. 3, top) and protein expression (Fig. 3, bottom) in 12 hours Compared with the control group, there was a statistically significant increase (p <0.01), and the amount of expression decreased in concentration-dependent manner after treatment with sacchinone. In the case of the expression level of the mRNA of the SREBP-1, when the expression level of the control group was 1, the T0901317 treatment group was 2.80, the sachinone 10 μM treatment group was 1.98, and the 30 μM treatment group was 1.31.

Experimental Example 4: Effect of Succinone Treatment on Weight Gain Due to High Fat Diet

Male C57BL / 6 mice (average body weight 25-30 g) were bred for 11 weeks on a high-fat diet (Dyets Inc., Bethlehem) or normal diet and for the last 5 weeks saucionone (10 or 30 mg / kg, 5 times / week). ) Was administered. The weight and condition of the animals were examined once a week and each group consisted of a total of 10 mice. No significant changes were observed in the amount of food and water for the duration of the experiment.

Administration of Succinone 30 mg / kg significantly reduced weight gain due to a high lipid diet (FIG. 4).

Experimental Example 5: Effect of Succinone Treatment on LXRα Increased Expression with High Lipid Diet

Various molecular and cellular markers were observed to elucidate the excellent synergistic treatment mechanism of fatty liver by sacchinone administration. First, the inhibitory effect of sacchinone on lipid-induced genes induced by high lipid diet In order to observe, the expression level of the intracellular lipid sensor LXRα was measured.             

LXRα expression in mice fed high-fat diet and normal diet for 11 weeks, and in the group fed high-fat diet, mice receiving sacchinone (10 or 30 mg / kg, 5 times / week) for the last 5 weeks. The degree was measured. After separating LXRα mRNA from liver tissue and synthesizing cDNA through RT-PCR, using specific primers (mouse LXR, 5'-TGCCATCAGCATCTTCTCTG-3 '(sense) and 5'-GGCTCACCAGCTTCATTAGC-3' (antisense)) Analysis was by real-time PCR. When the level of LXRα mRNA of the normal diet group (ND) is 1, the relative LXRα mRNA levels of the high-lipid diet group or the high-lipid diet and saucionone administration group are shown in FIG. 5.

The expression of LXRα, an intracellular lipid sensor, was significantly increased to 2.03 by the high-lipid diet (p <0.01), and the increase in LXRα expression was 1.66 at 10 mg / kg administration and 30 mg / kg administration at the same time. It was confirmed that it was suppressed to 1.53 (p <0.01) (Fig. 5 top).

Experimental Example 6: Effect of Succinone Treatment on SREBP-1 with High Lipid Diet

The expression of SREBP-1, a transcription factor that regulates lipogenesis enzyme gene expression, was also increased by a high-lipid diet, and the effect of Succinone administration was observed.

SREBP in mouse liver tissue fed 11 weeks on high-fat diet and normal diet, and in the group fed on high-fat diet, administration of sacchinone (10 or 30 mg / kg, 5 times / week) for the last 5 weeks. The degree of expression of -1 was measured. After isolation of SREBP-1 mRNA from liver tissue and synthesis of cDNA via RT-PCR, specific primers (mouse SREBP-1, 5'-AACGTCACTTCCAGCTAGAC-3 '(sense) and 5'-CCACTAAGGTGCCTACAGAGC-3' (antisense) ) Was analyzed by real-time PCR. When the level of the SREBP-1 mRNA of the normal diet group (ND) is 1, the relative SREBP-1 mRNA level of the high-lipid diet group or the high-lipid diet group and the sacchinone administration group is shown in FIG. 5.

The expression of SREBP-1 mRNA was significantly increased (p <0.01) by high-lipid diet (p <0.01), and the increase was 1.27 at 10 mg / kg and 1.31 at 30 mg / kg. Confirmed inhibition (p <0.01) (bottom of FIG. 5).

Experimental Example 7: Effect of Succinone Treatment on the Content of Triglycerides in Liver Tissue Accumulated by High Fat Diet

The content of triglycerides in liver tissue is an indicator of fatty liver. The mice were fed a normal diet and a high-lipid diet for 11 weeks and the last 5 weeks were administered 10 or 30 mg / kg of sacchinone five times per week. The effect was observed.

As a result of measuring triglyceride content in liver tissue after administration of sautinone, the triglyceride content in liver tissue was 89.22 mg / g compared to 20.55 mg / g liver in the normal diet group in mice fed with high-lipid diet for 11 weeks. iver. significantly increased in iver, 42.36 mg / g liver when 10 mg / kg of triglyceride content was increased by administration of high-lipid diet when saucinone was administered, and 32.39 mg / kg when 30 mg / kg was administered. g liver).

Experimental Example 8: Effect of Succinone Administration on Liver Tissue of Fatty Animal Model Induced by High Fat Diet

 Hepatic tissues were analyzed for fat-induced fatty liver treatment after Succinone administration using Oil Red O staining.

Liver tissue collected in Experimental Example 7 was fixed with 10% neutral formalin solution, and the tissue was embedded with paraffin after the usual fixation procedure and dehydration. The embedded tissues were tissue sections with a thickness of 4 μm, stained with oil red o, and observed with an optical microscope.

As a result, in the high-fat diet group, the red stained area was markedly red with oil red o (HFD), and in the high-fat diet group (HFD + Sau) to which the sautinone was administered, the red-stained area was significantly higher than the high-fat diet group. It was shown to have an excellent therapeutic effect (FIG. 7).

Experimental Example 9: ALT Measurement and H & E Staining During Succinone Treatment

Blood was collected from the mice treated in the same manner as Experimental Example 4-8, and the blood ALT (Alanine transaminase) was measured using Spectrum (automatic blood chemistry analyzer) (Abbott Laboratories, Abbott Park, IL). It was measured by.

ALT levels were significantly increased in the high-fat diet group to 185 IU / l compared to 43 IU / l in the normal diet group, and 92 and 106 in the high-fat diet group fed 10 mg / kg and 30 mg / kg of sacchinone, respectively. It was confirmed that IU / l was significantly reduced compared to the high lipid diet group (Fig. 8). As a result of H & E staining, hepatic necrosis (arrow) and infiltration of inflammatory cells (arrow head) and fat accumulation in the liver tissue (starred) were significantly reduced in comparison with the control group, and liver tissue damage due to fat accumulation was observed. And the inhibitory effect of inflammation could be confirmed (FIG. 9).

Iii. Effect of Triticale Extract

Experimental Example 10: LXRα activity inhibitory effect (LXRE binding capacity inhibition) effect by the three hundred seconds extract treatment

In order to observe the effect of the treatment of the extract of three hundred seconds containing sacchinone on LXRα activity, a reporter gene assay (Luciferase reporter gene assay) was carried out in the same manner as in Experiment 2.

After transfection of the vector containing LXRE into the HepG2 cell line, and after 12 hours, 10 μg / ml or 30 mM sachet extracted from Trichophyt extract (SE) extracted with chloroform containing 5% or 10% by weight of sacchinone in HepG2. The rice paddy was pretreated for 1 hour (10 μg / ml of 300 sec extract (SE) containing 10% of sacchinone contained 3 μM of sacchinone). Thereafter, LXRα activator T0901317 (10 M) was treated for 12 hours and luciferase activity was measured.

Luciferase activity was increased to 2.40 in the TO901317 treatment group when the activity of the control group was 1, and significantly decreased in the concentration range tested to about 1.24 or 1.13 when pre-treatment of the 300 sec extract containing 5% or 10% of sacchinone was performed. It was confirmed that (Fig. 10).

Experimental Example 11: Effect of three hundred sec extract treatment on weight gain due to high lipid diet

After induction of fatty liver with high fat diet (60% fat) for 4 weeks, the efficacy of the sacchinone-containing Trichoxa extract was measured by comparison with the control group administered with the vehicle and the experimental group administered with the drug for the next 4 weeks. Table 1 shows the substances administered to each animal group and the dose thereof in Experimental Examples 11 to 14, each group consisting of 10 mice.

Table 1

Fauna		Dietary Conditions	Dosing medication
One	ND	Normal diet	-
2	ND + SE		100 mg / kg extract of Trichophyte containing 10% sacchinone
3	HFD	High fat diet (60% fat)	-
4	HFD + SE 10		Triglyceride extract with 10% sacchinone 10 mg / kg
5	HFD + SE 30		30% extract of Trichoola extract containing 10% sacchinone
6	HFD + SE 100		100 mg / kg extract of Trichophyte containing 10% sacchinone
7	HFD + Sau		Succinone	10 mg / kg
8	HFD + Godex		Godex®	250 mg / kg

Male C57BL / 6 mice (average weight 25-30 g) were bred for 8 weeks on a 60% fat (Dyets Inc., Bethlehem) or normal diet and three hundred seconds containing 10% by weight of sacchinone for the last 4 weeks. Extracts (10, 30 or 100 mg / kg, 5 times a week) were administered. Succinone 10 mg / kg and Godex 250 mg / kg were used as positive controls, respectively. The weight and condition of the animals were checked once a week.

Administration of Trichophyte extract 10, 30, 100 mg / kg for 4 weeks significantly reduced the weight gain due to a high-fat diet (Fig. 11). No significant changes were observed in the amount of food and water for the duration of the experiment.

Experimental Example 12: Effect of Three hundred Second Extract Treatment on the Content of Triglycerides in Liver Tissue Accumulated by High Fat Diet

Liver tissue accumulated in a high-lipid diet after 8 weeks of normal and high-lipid diet in mice and 10, 30, or 100 mg / kg of trichophytium extract containing 10% by weight of sachinone for the last 4 weeks. The effect of the extract of triticale on triglyceride content was observed. Succinone 10 mg / kg and Godex 250 mg / kg were used as positive controls, respectively.

In mice fed with high-fat diet for 8 weeks, triglyceride content in liver tissue was significantly increased to 69.69 mg / g liver compared to 24.5 mg / g liver in normal diet. Here's a positive control, Godex^®To In the case of administration, the effect of inhibiting fat accumulation was found to be 57.09 mg / g liver, but in the group of 10, 30, 100 mg / kg tricot extract or 10 mg / kg of sautinone, respectively, 36.18 mg. , 24.39, 29,49, 26.74 mg / g liver, administration of high-lipid diets throughout the tested concentrations significantly reduced the amount of triglycerides in the tissues (FIG. 12).

Experimental Example 13: Effect of Administration of Trichophytaceae Extract on Liver Tissue of Fatty Animal Model Induced by High Fat Diet

Hepatic tissues were analyzed using the oil red o stain method, a fat-specific staining agent, after treatment with triglyceride extracts. The liver tissues collected in Experimental Example 11 were stained with oil red o in the same manner as in Experimental Example 8 and observed with an optical microscope. The results were calculated in five steps according to the degree of change and expressed as the Oil Red O index. (1 = no change; 2 = signs of pathological changes; 3 = mild pathological changes; 4 = moderate pathological changes; 5 = marked pathological changes)

As a result, in the high lipid diet group, the red stained portion was remarkable compared to the normal diet group, and fat was deposited in the liver tissue, and the red stained portion was significantly reduced in the trichophytium extract or the sachinone administration group. On the other hand, the oil red o index was 4.30 in the high-fat diet group when the normal diet was 1, and 2.45 and 2.05 in the case of 10, 30, 100 mg / kg of 300 sec extract or 10 mg / kg of sautinone, respectively. , 1.70, 2.33 were significantly lowered, indicating a decrease in fat content. On the other hand, the Godex-administered group showed a slight decrease in the oil red O index to 4.10, indicating that the three hundred sec extract of the present invention had an excellent therapeutic effect compared to the positive control group (FIGS. 13 and 14).

Experimental Example  14: When processing three hundred seconds extract ALT  Measurement and H & E Dyeing

Blood was collected from each group of mice treated as in Experimental Example 11-13, the blood ALT was measured using Spectrum (Abbott Laboratories, Abbott Park, IL), and liver tissues of the mice were collected to perform H & E staining. After observation under a microscope.

The ALT level was significantly increased to 152.1 IU / l compared to 51.75 IU / l in the high-fat diet group, and in the high-fat diet group administered 10, 30, 100 mg / kg and 10 mg / kg of sacchinone. It was confirmed that 81.3, 63.6, 60.9, 106.1 IU / l was significantly reduced compared to the high-fat diet group (Fig. 15). In addition, when H & E staining was observed, necrosis (arrow), infiltration of inflammatory cells (arrow head), and fat accumulation in the liver tissue (starred) were significantly decreased compared to the control group when administration of the extract of Trichophytica was observed. It was possible to confirm the treatment efficacy of liver damage and fatty hepatitis (FIG. 16).

 Hereinafter, the preparation examples of the composition containing the three hundred seconds extract or sacchinone of the present invention, but the present invention is not intended to limit it, but is intended to explain in detail only.

Formulation Example 1 Preparation of Powder

Succinone 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

Succinone 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

300 mg extract of the present invention

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

Succinone 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

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

Formulation Example 5 Preparation of Liquid

20 mg of triticale extract of the present invention

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method for preparing a liquid, each component is added to purified water to dissolve and mix, and an appropriate amount of lemon flavor is added. Purified water is added to adjust the total volume to 100 ml, and then filled into a brown bottle and sterilized to prepare a liquid.

Formulation Example 6 Preparation of Health Functional Food

1000 mg of three hundred seconds extract of the present invention

Vitamin mixture proper amount

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

Nicotinamide 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. Granules can be prepared and used in the manufacture of health foods according to conventional methods.

Formulation Example 7 Preparation of Healthy Drink

Succinone 100 mg

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 according to a conventional healthy beverage production method, and then 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 refrigerated and then stored in the present invention For the preparation of healthy beverage compositions.

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

Since the composition containing the tritical extract or sacchinone of the present invention as an active ingredient can be effectively used for the prevention, improvement and treatment of diseases caused by the overexpression or overactivity of LXRα or SREBP-1, fatty acid such as fatty liver Prevention, improvement and treatment of liver disease, hypertriglyceridemia, hyperreninemia, renin-induced hypertension, aldosteroneism, adrenal protein dystrophy, renal glomerulosis, proteinuria, kidney damage, etc. It can be used industrially for.

The attached Sequence Listing file is incorporated herein by reference.

Claims (9)

1. Liver X receptor (LXRα) or SREBP-1 (Sterol Response) comprising at least one selected from the group consisting of sacchinone, trichoola extract, pharmaceutically acceptable salts thereof, solvates thereof and hydrates thereof Pharmaceutical composition for the prevention or treatment of diseases caused by overexpression or overactivity of Element Binding Protein-1).
2. Sauuccinone, triticale extract. Hepatic diseases due to the accumulation of liver lipids, hypertriglyceridemia, hyperreninemia, renin, comprising at least one selected from the group consisting of pharmaceutically acceptable salts thereof, solvates thereof, and hydrates thereof as active ingredients. A pharmaceutical composition for preventing or treating a disease selected from the group consisting of hypertension, aldosteroneism, adrenal protein dystrophy, renal glomerulosclerosis, proteinuria and nephropathy.
3. The liver disease according to claim 2, wherein the liver disease due to the accumulation of liver lipids is selected from the group consisting of alcoholic fatty liver, nonalcoholic fatty liver, fatty hepatitis, liver fibrosis, liver cirrhosis, hepatomegaly, liver failure, liver failure, gallstones and portal hypertension Pharmaceutical composition.
4. Prevention of diseases due to overexpression or overactivity of LXRα or SREBP-1, comprising as an active ingredient at least one selected from the group consisting of sautinone, triticale extract, salts thereof, solvates thereof and hydrates thereof; and Health functional food for improvement.
5. 5. The liver disease according to claim 4, wherein the disease is alcoholic fatty liver, nonalcoholic fatty liver, fatty hepatitis, liver fibrosis, liver cirrhosis, hepatomegaly, liver failure, liver failure, gallstones and hepatic lipid hypertension. Health functional food is selected from the group consisting of hypertriglyceridemia, hyperreninemia, hypertension caused by renin, aldosteroneism, adrenal protein dystrophy, renal glomerulus, proteinuria and kidney damage.
6. A dietary supplement according to claim 4 or 5 in the form of a powder, granule, tablet, capsule, syrup or beverage.
7. A method for producing an extract of three hundred seconds, comprising extracting at 10 to 100 ° C. by adding a low polar solvent, alcohol or a mixture thereof to three hundred seconds.
8. The method according to claim 7, further comprising the step of removing the extract containing the non-polar component by extracting three hundred seconds with a non-polar solvent at 10 to 100 ℃ before said step.
9. The method of claim 7 or 8, wherein the nonpolar solvent is selected from the group consisting of n-hexane, cyclohexane, benzene and petroleum ether, and the low polar solvent is selected from the group consisting of chloroform, dichloromethane, ether and ethyl acetate. The production method selected.

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