WO2021242043A1 - COMPOSITION FOR PREVENTING, TREATING OR AMELIORATING METABOLIC SYNDROME COMPRISING LACTOBACILLUS FERMENTED PRODUCT OF PEONY EXTRACT OR β-GENTIOBIOSYL PAEONIFLORIN AS ACTIVE INGREDIENT - Google Patents

Abstract

The present invention relates to a composition for preventing, treating or ameliorating a metabolic syndrome, comprising a lactobacillus fermented product of a peony extract or β-gentiobiosyl paeoniflorin as an active ingredient and, more specifically, to a composition for preventing, treating or ameliorating a metabolic syndrome, comprising as an active ingredient a lactobacillus fermented product of a peony extract or β-gentiobiosyl paeoniflorin, which exhibits the action of increasing glucose uptake into muscle cells and inhibiting the activity of lipase.

Description

Composition for preventing, treating or improving metabolic syndrome comprising lactic acid bacteria fermented product of peony extract or β-gentiobiosil paoniflorin as an active ingredient

This application claims priority to Korean Patent Application No. 10-2020-0065497, filed on May 29, 2020, and Korean Patent Application No. 10-2020-0068568, filed on June 5, 2020, and the above specification The entirety of this application is incorporated herein by reference.

The present invention relates to a composition for preventing, treating, or improving metabolic syndrome comprising a fermented product of lactic acid bacteria of a peony extract or β-gentiobiosil paoniflorin as an active ingredient, and more particularly, to a composition for improving muscle cells. It relates to a composition for preventing, treating or improving metabolic syndrome, comprising as an active ingredient a lactic acid bacterium fermented product of a peony extract or β-gentiobiosil paoniflorin, which increases glucose absorption and inhibits the activity of lipase.

In modern society, the rapid economic development and abundant nutritional intake, while significantly decreasing the amount of exercise, causes two or more complex metabolic diseases such as obesity, diabetes, high blood pressure, hypertriglyceridemia, hypercholesterolemia, and arteriosclerosis. The prevalence of Metabolic Syndrome is increasing, and according to the 2005 National Health and Nutrition Examination Survey data, the overall figure is 32.3% (32.9% for men, 31.8% for women). As a result, heart disease and stroke are increasing enough to occupy the second and third leading causes of death in Koreans. This is a symptom that occurs when the metabolic wastes and toxins that are caused by a poorly balanced metabolism cannot be released, and the accumulated waste products in the body lose their functions. Metabolism also known as insulin resistance syndrome syndrome develops. Metabolic syndrome can cause damage to the coronary arteries, which can cause heart disease or stroke, or decrease the ability of the kidneys to remove salts to cause high blood pressure, increase the proportion of triglycerides that cause cardiovascular disease, and increase blood pressure. It increases the risk of clotting and is also known to result in reduced insulin production with type 2 diabetes, resulting in damage to the eyes, kidneys, and nerves.

An effective drug for the treatment of metabolic syndrome has not yet been developed, and only attempts are made to treat the metabolic syndrome using drugs for diabetes, dyslipidemia and hypertension. Currently available drugs for the treatment of metabolic syndrome include metformin, a thiazolidinediones (TZD) class of drugs used as a treatment for diabetes, a glucosidase inhibitor, and a dipeptidyl peptidase (DDP)-IV inhibitor. In addition, blood pressure drugs and dyslipidemia drugs are attracting attention. However, there is a limit to improving the metabolic syndrome with these drugs.

Looking at the known factors related to the cause and treatment of metabolic syndrome, exercise, diet, body weight, blood sugar, triglyceride, cholesterol, insulin resistance, adiponectin, leptin, AMP-activated protein kinase (AMPK) activity , sex hormones such as estrogen, genetic factors, and the concentration of malonyl-CoA in vivo are directly or indirectly involved.

Therefore, for the effective management or treatment of metabolic syndrome with complex symptoms, it is ideal to develop a material capable of improving obesity, which is a fundamental cause of metabolic syndrome, at the same time as having a hypoglycemic effect for maintaining normal blood sugar. Research and development for therapeutic agents is insufficient.

On the other hand, the peony ( Paeonia lactiflora ) is a perennial plant of the buttercup family, mainly grown at the foot of a mountain or in a valley. Several stems come out of one stem and stand upright, about 60cm high, and there are no hairs on the leaves and stems. Although there are several roots, it is thick and thin in the shape of a pointed cylinder with long both ends. One flower blooms at the tip of the stem from May to June, and it is large and beautiful, and the cultivated one is about 10cm in diameter. The flower color is red and white, and there are many horticultural varieties. The root is used as medicine for pain relief, abdominal pain, menstrual pain, amenorrhea, hematemesis, anemia, and bruises. In addition, when peony root is used as a drug, it can be divided into red peony and earl peony according to the pre-processing method. Red peony is dried by removing the root of the beard, and in the case of white peony, the root is removed and boiled in boiling water, then the outer skin is removed or boiled again.

Meanwhile, Korean Patent Application Laid-Open No. 10-2009-0109446 discloses a pharmaceutical composition for preventing and treating obesity of a complex herbal extract containing peony, and paeoniflorin is known as an indicator material of peony or peony extract. , their anti-obesity-related activity was not sufficiently limited to exhibit physiologically effective effects.

Therefore, the present inventors were studying to find an effective method for the treatment of obesity by metabolizing or improving the peony extract, and when fermenting the peony extract with Leuconostoc sp. LN180020 (KCTC 13719BP) Oniflorin is converted to another material, and the lactic acid bacteria fermented product of the peony extract containing it has significantly improved the activity of improving metabolic syndrome compared to the peony extract, thereby completing the present invention. In addition, paeoniflorin, one of the active components of the peony extract, is β-gentiobiosyl paeoniflorin by fermentation of Leuconostoc sp. LN180020 (KCTC 13719BP). ), which confirmed that it exhibits significantly improved metabolic syndrome preventive or therapeutic activity compared to paoniflorin, thereby completing the present invention.

Therefore, it is an object of the present invention to prevent metabolic syndrome comprising a lactic acid bacterium fermented product or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof as an active ingredient of a peony (Paeonia lactiflora) extract Or to provide a pharmaceutical composition for treatment.

In addition, an object of the present invention is a lactic acid bacteria fermented product of peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin (β-gentiobiosyl paeoniflorin) or a pharmaceutical for preventing or treating metabolic syndrome consisting of a pharmaceutically acceptable salt thereof It is to provide an enemy composition.

In addition, it is an object of the present invention to prevent or treat metabolic syndrome consisting essentially of lactic acid bacteria fermented product of Paeonia lactiflora extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof It is to provide a pharmaceutical composition for.

Another object of the present invention is to prevent or prevent metabolic syndrome comprising a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof as an active ingredient. It is to provide a food composition for improvement.

In addition, another object of the present invention is for the prevention or improvement of metabolic syndrome consisting of lactic acid bacteria fermented product of peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof To provide a food composition.

In addition, another object of the present invention is to prevent metabolic syndrome consisting essentially of lactic acid bacteria fermented product of Paeonia lactiflora extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. It is to provide a food composition for improvement.

Another object of the present invention is the lactic acid fermented product or β-gentiobiosyl paeoniflorin of an extract of Paeonia lactiflora for manufacturing a preparation for the treatment of metabolic syndrome or a pharmaceutically acceptable salt thereof. to provide use.

Another object of the present invention is to obtain an effective amount of a preparation containing a lactic acid bacterium fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. It is to provide a method for treating metabolic syndrome comprising administering to an individual.

In order to achieve the above object, the present invention is a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof as an active ingredient. It provides a pharmaceutical composition for preventing or treating metabolic syndrome, including.

In addition, the present invention provides a pharmaceutical composition for preventing or treating metabolic syndrome comprising a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. provides

In addition, the present invention is a pharmaceutical for preventing or treating metabolic syndrome consisting essentially of a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. Provides an enemy composition.

In order to achieve another object of the present invention, the present invention is a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin (β-gentiobiosyl paeoniflorin) or a pharmaceutically acceptable salt thereof as an active ingredient. It provides a food composition for preventing or improving metabolic syndrome comprising a.

In addition, the present invention provides a food composition for preventing or improving metabolic syndrome comprising a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. to provide.

In addition, the present invention is a food for preventing or improving metabolic syndrome consisting essentially of lactic acid bacteria fermented product of Paeonia lactiflora extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof A composition is provided.

In order to achieve another object of the present invention, the present invention provides a lactic acid bacterium fermented product of Paeonia lactiflora extract or β-gentiobiosyl paeoniflorin or its Provided is the use of a pharmaceutically acceptable salt.

In order to achieve another object of the present invention, the present invention is a lactic acid bacteria fermented product of peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin (β-gentiobiosyl paeoniflorin) or a pharmaceutically acceptable salt thereof. Provided is a method for treating metabolic syndrome, comprising administering an effective amount of the agent to a subject in need thereof.

Hereinafter, the present invention will be described in detail.

The present invention relates to a pharmaceutical for preventing or treating metabolic syndrome comprising, as an active ingredient, a lactic acid bacteria fermented product of a peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof. Provides an enemy composition.

As used herein, the term 'peony' refers to the root, stem, leaf, flower, seed, and fruit of a plant having the scientific name Paeonia lactiflora , preferably the root of Paeonia lactiflora used as a medicine. The peony as a medicine is also called Paeoniae Radix or Peony root, and more specifically, it refers to the root of the peony Paeonia lactiflora Pallas or other closely related plants (Peony and Paeoniaceae) used as medicinal parts.

The peony extract may be extracted by a solvent extraction method known in the art. The extraction solvent is not limited thereto, but water, a lower alcohol having 1 (C1) to 6 (C6) carbon atoms, an organic solvent, or a mixture thereof may be used. The lower alcohol having 1 (C1) to 6 (C6) carbon atoms is not limited thereto, but may be methanol, ethanol, alcohol, propanol, isopropanol, butanol, pentanol, or hexanol, and the organic solvent is not limited thereto, acetone, ethyl acetate, n-nucleic acid, diethyl ether, acetone, or benzene. Preferably, extraction may be performed with water or an alcohol having 1 to 6 carbon atoms or a mixed solvent thereof, and more preferably water, ethanol or alcohol. A small amount of water (eg, in the form of 95% ethanol or 99% ethanol) may be added to alcohol to reflect physicochemical properties such as hygroscopicity.

The extraction temperature and time are not particularly limited as long as the purpose of extraction of the active ingredient is achieved, but extraction may be performed at 30° C. to 121° C. for 10 minutes to 12 hours. In order to facilitate extraction of the active ingredient during extraction, it may be extracted by pressing. Meanwhile, the extraction may be performed by supercritical extraction or subcritical extraction using an appropriate solvent such as carbon dioxide.

In the present invention, the 'lactic acid bacteria' may be Leuconostoc citreum or Leuconostoc sp. LN180020 having an accession number of KCTC 13719BP, preferably Leuconostoc genus having an accession number of KCTC 13719BP ( Leuconostoc sp.) LN180020. The 'lactic acid bacteria' is characterized in that it has a bioconversion ability to convert paoniflorin.

In an embodiment of the present invention, vegetable lactic acid bacteria derived from kimchi were isolated in order to search for useful lactic acid bacteria. As a result, the genus Lactobacillus ( Lactobacillus ), the genus Petiococcus ( Pediococcus ), the genus Weissella , the genus Leuconostoc ( Leuconostoc ) and the genus Enterococcus ( Enterococcus ) 193 of 39 species (species) included strains were isolated.

In one embodiment of the present invention, the isolated lactic acid bacteria strain was inoculated into a hot water extract of peony and fermented to prepare a lactic acid bacteria fermented product of the peony extract, and a strain into which paeoniflorin was bioconverted in the fermented product was searched. As a result, it was confirmed that the peak around 15 minutes, which is expected to be specifically paoniflorin, decreased in one of the isolated strains, and the peak of the new material increased at 14.2 minutes.

This strain was identified and named as Leuconostoc sp. LN180020 strain, and was deposited at the Center for Biological Resources (KCTC) on November 20, 2018 (Accession No.: KCTC 13719BP).

In one embodiment of the present invention, whether the fermented product of LN180020 in the leuconostok genus of the peony extract exhibits the activity of inhibiting lipase decomposing lipids, and the degree was compared with that of a non-fermented general peony extract. . As a result, the fermented product of lactic acid bacteria of the peony extract showed very excellent lipase inhibitory activity, and it was confirmed that this activity was significantly superior to that of the general peony extract. That is, it indicates that the lactic acid bacteria fermented product of the peony extract has very excellent anti-obesity activity.

In another embodiment of the present invention, as a result of comparing the effects of lactic acid bacteria fermented peony extract or general peony extract to an animal model induced by administration of a high-fat diet, weight, glycated hemoglobin, fasting blood sugar, Total cholesterol, triglyceride, and LDL levels in blood, as well as AST and ALT levels, were significantly decreased in the fermented product group compared to the general peony extract group. It was confirmed that there was a significant increase in That is, the fermented lactic acid bacteria of the peony extract shows that the metabolic syndrome improvement effect is significantly higher than that of the general peony extract.

In the present invention, the β-gentiobiosyl paoniflorin is a glycoside of paoniflorin having the structure of Formula 1 below, and is included in the scope of the present invention or a pharmaceutically acceptable salt thereof. β-Genthiobiosyl paoniflorin of Formula 1 or a pharmaceutically acceptable salt thereof may be chemically synthesized by a method known in the art, commercially available materials, or isolated from natural products.

[Formula 1]

In one aspect of the present invention, the β- gentiobiosyl paoniflorin is Leuconostoc sp. LN180020 strain (Accession No.: KCTC13719BP) characterized in that it is produced by bioconversion of paoniflorin can do.

In another aspect of the present invention, the β- gentiobiosil paoniflorin is leuconostok in powder, extract or extract of peony ( Paeonia lactiflora ) or genus Paeonia ( Paeonia ) containing paoniflorin as an index component. Genus ( Leuconostoc sp.) LN180020 strain (Accession No.: KCTC13719BP) can be characterized in that it is produced by inoculation and then fermentation.

In the present invention, the term “pharmaceutically acceptable salt” refers to a salt in a form that can be used pharmaceutically among salts that are substances in which cations and anions are combined by electrostatic attraction, usually with metal salts, organic bases and salts, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. For example, the metal salt may be an alkali metal salt (sodium salt, potassium salt, etc.), alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), an aluminum salt or the like; Salts with organic bases include triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylenediamine salts with, etc.; salts with inorganic acids may be salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like; Salts with organic acids may be salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like; Salts with basic amino acids may be salts with arginine, lysine, ornithine and the like; The salt with an acidic amino acid may be a salt with aspartic acid, glutamic acid, or the like. For the purposes of the present invention, the pharmaceutically acceptable salt may be interpreted as an acid addition salt or a base addition salt of imatinib suitable for the treatment of patients expected to develop vascular permeability-related diseases or have the disease, but in particular, However, the present invention is not limited thereto.

In an example of the present invention, whether β-gentiobiosyl paoniflorin exhibits an activity of inhibiting lipase decomposing lipids, and the extent thereof was compared with paoniflorin. As a result, β-gentiobiosyl paoniflorin exhibited very excellent lipase inhibitory activity, and it was confirmed that this activity was significantly superior to paoniflorin. That is, it indicates that β-gentiobiosyl paoniflorin has very excellent anti-obesity activity.

In another embodiment of the present invention, as a result of comparing the effects of administration of β-gentiobiosil paoniflorin or paoniflorin to an animal model induced by administration of a high-fat diet, body weight, total cholesterol in blood , triglycerides and LDL levels, AST, ALT levels, fasting blood glucose and insulin resistance were significantly decreased in the β-gentiobiosil paoniflorin group compared to the paoniflorin group, and even in the case of β- It was confirmed that a significant increase was observed in the gentiobiosil paoniflorin-administered group. That is, β-gentiobiosyl paoniflorin shows that the improvement of metabolic syndrome including diabetes, obesity, fatty liver and dyslipidemia is significantly higher than that of paoniflorin.

As used herein, 'treatment' or 'improvement' refers to inhibiting the occurrence or recurrence of a disease, alleviating symptoms, reducing direct or indirect pathological consequences of a disease, reducing the rate of disease progression, ameliorating, ameliorating, alleviating or ameliorating a disease state. means the prognosis. As used herein, the term 'prevention' refers to any action that suppresses the onset or delays the progression of a disease.

In the present invention, the metabolic syndrome may be one or more selected from the group consisting of diabetes, obesity, fatty liver, hyperlipidemia, arteriosclerosis, insulin resistance syndrome, and complications thereof, and preferably includes diabetes and/or obesity, and complications thereof. may be doing

In the present invention, 'diabetes', which is the subject of treatment or prevention, is a metabolic disorder syndrome characterized by a deficiency of insulin hormone produced in beta cells of the pancreas or abnormal insulin resistance and hyperglycemia caused by defects of both of these. Such diabetes can be divided into insulin-dependent diabetes mellitus (IDDM, Type 1) and non-insulin-dependent diabetes mellitus (NIDDM, Type 2) caused by insulin resistance and impaired insulin secretion. In both type 1 and type 2 diabetes, various complications such as heart disease, intestinal disease, eye disease, neurological disease, and stroke occur. Short-term hypoglycemia and hyperglycemia will cause acute complications. Diabetes mellitus causes chronic high blood sugar and lipid and protein metabolism as well as carbohydrate metabolism. The conditions are various and are directly caused by hyperglycemia, including diabetic peripheral neuropathy, diabetic retinopathy, diabetic nephropathy, diabetic cataract, keratosis, diabetic arteriosclerosis, etc. in the retina, kidney, nerve, and cardiovascular system.

Accordingly, diabetes in the present invention includes diabetes or diabetes complications caused by diabetes, and more specifically, may be type 1 diabetes, type 2 diabetes, or gestational diabetes. In the case of diabetic complications, it may be a disease caused by the destruction or functional decline of islet cells of Langerhans, or a disease caused by an increase in blood glucose concentration, specifically, but not limited to, diabetic peripheral neuropathy, diabetic retinopathy, diabetic nephropathy, diabetic cataract, keratosis, diabetic arteriosclerosis.

Since the composition provided by the present invention exhibits an action of increasing glucose absorption into muscle cells, it can be very usefully utilized for glycemic control in diabetic patients.

In addition, the composition provided by the present invention exhibits an activity of inhibiting lipase, which aids in absorption by decomposing fat, and thus can be very usefully used to improve obesity and metabolic syndrome induced thereby.

In general, people who are overweight, but not obese, have a lot of muscle because people with a lot of muscle can gain a lot of weight, so the condition of excessive fat tissue in the body is referred to as 'obesity'. According to the World Health Organization (WHO), the clinical body mass index is 25 in Korea, and 30 or more according to the World Health Organization (WHO). Obesity is diagnosed when the ratio is high, and it refers to a disease that occurs in both adults and children.Such obesity is not only weight gain, but also overeating, binge drinking and bulimia, hypertension, diabetes, increased plasma insulin concentration, insulin resistance, and hyperlipidemia. , metabolic syndrome, insulin resistance syndrome, obesity-related gastroesophageal reflux disease, arteriosclerosis, hypercholesterolemia, hyperuric acidemia, lower back pain, cardiac hypertrophy and left ventricular hypertrophy, lipodystrophy, nonalcoholic steatohepatitis, cardiovascular disease or polycystic ovarian syndrome Therefore, when using the composition according to the present invention, the prevention or treatment of obesity-related diseases as well as obesity can be achieved at the same time, and the treatment target for these obesity-related diseases is to reduce weight. Objects with desires are also included.

In the present invention, the 'fatty liver' is a disease caused by the accumulation of excessive fat (mainly, triglycerides) in the liver, and generally means that more than 5% of the weight of the liver is accumulated. In the present invention, the fatty liver includes non-alcoholic fatty liver, alcoholic fatty liver, and non-alcoholic fatty liver disease.

Preferably, in the present invention, the fatty liver includes non-alcoholic fatty liver disease (NAFLD). The non-alcoholic fatty liver disease (NAFLD) is a generic term for diseases caused by the deposition of fat in the liver of a person without a drinking history.

Specifically, the non-alcoholic fatty liver disease (NAFLD), regardless of alcohol, is overweight and obesity, insulin resistance, diabetes (type 1 diabetes (T1D), type 2 diabetes, etc.), dyslipidemia (hyperlipidemia), metabolic syndrome (For example, high blood pressure, high blood sugar, abdominal obesity, metabolic syndrome caused by low HDL cholesterol and high triglycerides) is a common chronic liver disease that is caused by the accumulation of excess fat (mainly, triglycerides) in the liver. This means that more than 5% of the liver's weight is accumulated as fat and diseases caused by it.

The non-alcoholic fatty liver disease (NAFLD) is a non-alcoholic fatty liver, non-alcoholic steatohepatitis (NASH), liver fibrosis (liver fibrosis) in which only fat is accumulated in the liver without damage to the liver cells. cirrhosis) may even develop into liver cancer. Specifically, the non-alcoholic fatty liver disease (NAFLD) is a simple fatty liver with only steatosis without hepatocellular damage, steatosis and type 2 steatohepatitis with an inflammatory reaction in the lobules due to severe and prolonged damage to the hepatocytes, steatosis and It can be divided into type 3 with fatty necrosis with balloon-shaped degeneration of hepatocytes and type 4 with mallysosomes or fibrosis with the above-mentioned type 3 findings.

The steatohepatitis is known as steatohepatitis in about 10% of all fatty liver disease patients, and the nonalcoholic steatohepatitis (NASH) is caused by an increase in triglyceride production due to hyperinsulinemia and a decrease in fat metabolism function. If the non-alcoholic fatty liver and non-alcoholic steatohepatitis are left unattended, liver function may deteriorate and liver cirrhosis may occur through liver fibrosis.

The liver fibrosis is a kind of wound healing process that appears in liver tissue. When the liver tissue is damaged by hepatitis virus, nonalcoholic fatty liver, drugs, etc., the vascular structure in the liver lobules in the process of continuously repeating damage and regeneration of the liver cells. is collapsed, and the regenerative nodules that have lost their function take their place, causing a fibrotic reaction in the liver, which can lead to cirrhosis, in which the liver becomes hard.

The damaged hepatocytes secrete reactive oxygen species and inflammatory substances to activate Kupffer cells and inflammatory cells, which leads to activation of hepatic stellate cells. In the normal state, hepatic stellate cells are only the main storage of vitamin A, but when liver damage occurs, they are activated and synthesize and secrete various extracellular matrixes such as collagen, which can cause liver fibrosis. If liver damage is chronically repeated, damaged hepatocytes are no longer regenerated and are gradually replaced by extracellular matrix such as collagen, which can lead to liver fibrosis and cirrhosis.

That is, the nonalcoholic fatty liver disease (NAFLD) is a continuous disease that progresses from simple nonalcoholic fatty liver to steatohepatitis and liver fibrosis to cirrhosis.

The pharmaceutical composition according to the present invention can be effectively used for the treatment of nonalcoholic fatty liver disease (NAFLD), not only nonalcoholic fatty liver, but also nonalcoholic steatohepatitis (NASH) and liver fibrosis.

In the present invention, 'dyslipidemia' refers to a state in which total cholesterol, LDL cholesterol, and triglycerides in the blood are increased or HDL cholesterol is decreased, and in the group consisting of hyperlipidemia, hypercholesterolemia and hypertriglyceridemia It may be any one or more selected. The hyperlipidemia refers to a state in which lipids including cholesterol and triglycerides are increased in blood. Hyperlipidemia, although asymptomatic, may increase the risk of coronary artery disease, such as atherosclerosis or myocardial infarction. The hypercholesterolemia is a state in which cholesterol in the blood is increased, and total cholesterol and LDL cholesterol are high. The hypertriglyceridemia refers to a state in which triglycerides are increased in blood. If dyslipidemia persists, excess cholesterol in the blood may be deposited on the walls of blood vessels. After a lot of cholesterol is deposited on the walls of blood vessels, inflammatory and oxidative reactions progress, reducing the elasticity of blood vessels, thickening blood vessel walls, and blocking smooth blood flow, complications such as arteriosclerosis, angina, and stroke may occur.

In the present invention, 'insulin resistance syndrome' is a concept that collectively refers to diseases induced by insulin resistance. Cell resistance to insulin action, hyperinsulinemia, increase in very low density lipoprotein (VLDL) and triglycerides, It refers to a disease characterized by a decrease in high density lipoprotein (HDL) and hypertension, and is a concept recognized as a risk factor for cardiovascular disease and type 2 diabetes (Reaven GM., Role of insulin resistance in human). disease, Diabetes, 37:1595-607 (1988)). It is also known that insulin resistance increases intracellular oxidative stress along with risk factors such as high blood pressure, diabetes, and smoking, and induces an inflammatory response by changing the signaling system to advance atherosclerosis.

The pharmaceutical composition according to the present invention may be formulated in various ways according to the route of administration by a method known in the art together with a pharmaceutically acceptable carrier for the prevention or treatment of metabolic syndrome. The carrier includes all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes.

The pharmaceutical composition according to the present invention may be administered to a patient in a pharmaceutically effective amount, that is, in an amount sufficient to prevent or alleviate symptoms and treat metabolic syndrome. For example, a typical daily dose may be administered in the range of about 0.01 to 1000 mg/kg, preferably, it may be administered in the range of about 1 to 100 mg/kg. The pharmaceutical composition of the present invention may be administered once or divided into several doses within a preferred dosage range. In addition, the dosage of the pharmaceutical composition according to the present invention may be appropriately selected by those skilled in the art according to the route of administration, administration, age, sex, body weight, individual differences, and disease state.

The route of administration may be oral or parenteral administration. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, rectal, or pancreatic administration. It may be administered internally, but is not limited thereto, and most preferably, it may be administered orally.

When the pharmaceutical composition of the present invention is orally administered, powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art together with a suitable oral administration carrier It can be formulated in the form of, etc. Examples of suitable carriers include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches, including corn starch, wheat starch, rice starch and potato starch, cellulose, Cellulose, including methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose, and the like, fillers such as gelatin, polyvinylpyrrolidone, and the like may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant if necessary. Furthermore, the pharmaceutical composition may further include an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative.

In addition, when administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administration agents, nasal inhalants, etc. together with suitable parenteral carriers.

In addition, the pharmaceutical composition may be administered by any device capable of transporting the active agent to a target cell. Preferred administration methods and formulations are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, or drip injections. Injections include aqueous solvents such as physiological saline or Ringer's solution, vegetable oil, higher fatty acid esters (eg, ethyl oleate), and non-aqueous solvents such as alcohols (eg, ethanol, benzyl alcohol, propylene glycol or glycerin). It can be prepared using, stabilizers for preventing deterioration (eg, ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH adjustment, to inhibit the growth of microorganisms and a pharmaceutical carrier such as a preservative (eg, phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

In the case of the injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injection, examples of suitable carriers include, but are not limited to, water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof, and/or a solvent or dispersion medium containing vegetable oil. can More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with triethanolamine or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, in most cases, the injection may further contain an isotonic agent such as sugar or sodium chloride.

As other pharmaceutically acceptable carriers, those known in the art may be referred to.

The pharmaceutical composition according to the present invention may contain one or more buffers (eg saline or PBS), carbohydrates (eg glucose, mannose, sucrose or dextran), antioxidants, bacteriostats, chelating agents (eg EDTA or glutathione), adjuvants (eg, aluminum hydroxide), suspending agents, thickening agents and/or preservatives.

In addition, the pharmaceutical compositions of the present invention may be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

In addition, the pharmaceutical composition of the present invention may be administered alone or in combination with a known compound having an effect on the treatment of metabolic syndrome.

On the other hand, the present invention is a lactic acid bacteria fermented product of peony (Paeonia lactiflora) extract or β-gentiobiosyl paeoniflorin (β-gentiobiosyl paeoniflorin) or a pharmaceutically acceptable salt thereof as an active ingredient to prevent or improve metabolic syndrome comprising It provides a food composition for use.

The food composition of the present invention includes all types of functional foods, nutritional supplements, health foods, and food additives.

Food compositions of this type can be prepared in various forms according to conventional methods known in the art. Although not limited thereto, for example, as a health food, the lactic acid bacteria fermented product of the peony extract can be prepared in the form of tea, juice, and drink and liquefied, granulated, encapsulated and powdered so that it can be consumed. In addition, it can be prepared in the form of a composition by mixing the lactic acid bacteria fermented product of the peony extract and a known active ingredient known to be effective in obesity. In addition, functional foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat, and processed foods (eg, ham, sausages) corn beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine, vegetable It can be prepared by adding the lactic acid bacteria fermented product of the peony extract to protein, retort food, frozen food, and various seasonings (eg, soybean paste, soy sauce, sauce, etc.). In addition, in order to use the lactic acid bacteria fermented product of the peony extract in the form of an additive, it can be prepared and used in the form of a powder or a concentrate.

The preferred content of lactic acid bacteria fermented product of peony extract or β-gentiobiosil paoniflorin in the food composition of the present invention is not limited thereto, but is preferably 0.1 to 90% by weight of the finally prepared food. More preferably, the food composition containing the lactic acid bacteria fermented product of the peony extract of the present invention or β-gentiobiosil paoniflorin as an active ingredient is mixed with an active ingredient known to be effective for metabolic syndrome, in particular, for health It may be prepared in the form of a functional food or dietary supplement.

The health functional food composition for preventing or improving metabolic syndrome of the present invention exhibits lipase inhibitory activity that promotes digestion and absorption by decomposing fat, and lactic acid bacteria fermented product or β-gen of peony extract that increases glucose absorption into muscle cells It contains thiobiosyl paoniflorin as an active ingredient, so it is characterized in that it is effective in preventing or improving metabolic syndrome.

On the other hand, in one embodiment of the present invention, LN180020 lactic acid bacteria in Leuconostok was identified through 16S rRNA sequencing analysis, and a new substance that showed an increase in anti-obesity activity was to be identified. As a result, LN180020 lactic acid bacteria of the genus Leuconostok showed the highest 16S rRNA sequence homology with Leuconostok citreum , and paeoniflorin with a molecular weight of 480.3, which is an indicator material of the peony extract, decreased, and a molecular weight of 642.3. It was found that bioconversion into β-gentiobiosyl paoniflorin.

In addition, the present invention relates to the use of a lactic acid fermented product of an extract of Paeonia lactiflora or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof for preparing a preparation for the treatment of metabolic syndrome to provide.

The present invention provides an effective amount of a formulation containing a lactic acid bacteria fermented product of Paeonia lactiflora extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof to an individual in need thereof. It provides a method for treating metabolic syndrome comprising administering.

The 'effective amount' of the present invention refers to an amount that exhibits the effect of improving, treating, preventing, detecting, diagnosing, or inhibiting or reducing metabolic syndrome when administered to an individual, and the 'individual' refers to an animal, preferably It may be an animal including a mammal, particularly a human, and may be an animal-derived cell, tissue, organ, or the like. The subject may be a patient in need of the effect.

The 'treatment' of the present invention refers to alleviating metabolic syndrome or symptoms of metabolic syndrome, which may include curing, substantially preventing, or ameliorating the condition of the disease, and including, but not limited to, alleviating, curing or preventing one or most of the symptoms resulting from

As used herein, the term “comprising” is used in the same sense as “including” or “characterized by”, and in the composition or method according to the present invention, specifically Additional components or method steps that have not been excluded are not excluded. In addition, the term “consisting of” means excluding additional elements, steps, or ingredients not specifically described. The term “essentially consisting of” means that, in the scope of a composition or method, it may include substances or steps that do not substantially affect the basic properties thereof in addition to the substances or steps described.

Therefore, the fermented peony extract lactic acid bacteria or β-gentiobiosil paoniflorin of the present invention has excellent lipase inhibitory activity, which promotes digestion and absorption by decomposing lipids, and promotes glucose absorption into muscle cells Therefore, it is expected that it can be usefully used in the development of foods and drugs for the improvement or treatment of metabolic syndrome.

1A and 1B are HPLC chromatograms showing the content of indicator substances and new useful substances in non-fermented peony extract (non-fermented, (FIG. 1A)) and lactic acid bacteria fermented product (fermented, (FIG. 1B)) of the peony extract. .

Figure 2 is a leuconostoc genus (Leuconostoc sp.) is a phylogenetic diagram analyzing the lineage of LN180020 (KCTC 13719BP).

Figure 3 is a non-fermented peony extract (non-fermented) and lactic acid bacteria fermented (fermented) of the peony extract, respectively, the peony indicator material before bioconversion (paeoniflorin, paeoniflorin) and the material after bioconversion to separate and purify the LC-MS is the result of performing

4 is a result showing H, C, HMBC NMR spectral data of substances before and after bioconversion in peony extract.

5 is a result of confirming the cell viability (cell viability) of the lactic acid bacteria fermented product of the peony extract and the non-fermented peony extract.

Figure 6 is the result of confirming the glucose uptake rate (relative glucose uptake) of the fermented product of Leuconostoc sp. LN180020 (KCTC 13719BP) and non-fermented peony extract of the peony extract into muscle cells.

Figure 7 is Lactobacillus plantarum subsp. plantarum of peony extract (Lactobacillus plantarum subsp. plantarum; Df2) LN180155 or Lactobacillus paraplantarum (Lactobacillus paraplantarum KACC 12373; Df3) fermented product and muscle cells of non-fermented peony extract This is the result of confirming the relative glucose uptake.

8 is a result of evaluating the lipase inhibitory activity of a non-fermented peony extract (non-fermented), lactic acid bacteria fermented product (fermented) of the peony extract, and a positive control group (olistat).

9A to 9D are results of measuring body weight, HbA1c, fasting blood sugar and HOMA-IR after administration of hot water peony extract or fermented extract of peony extract to a high-fat diet animal model.

10 is a result showing the results of measuring the indicated indicators by collecting blood after administering a hot water peony extract or a fermented extract of a peony extract to a high-fat diet animal model.

11A to 11D are results of measuring body weight, HbA1c, fasting blood glucose and HOMA-IR after administration of paoniflorin or β-gentiobiosil paoniflorin to a high-fat diet animal model.

12 is a result showing the results of measuring the indicated indicators by collecting blood after administration of paoniflorin or β-gentiobiosil paoniflorin to a high-fat diet animal model.

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

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

Example 1: Isolation of plant lactic acid bacteria from kimchi

To isolate vegetable lactic acid bacteria derived from kimchi, various kimchi source samples are gradually diluted with sterile water, spread on MRS (Man Rogosa and Sharpe) medium, and cultured at 28°C for 2 days. The strain isolated through 16s rRNA sequence analysis was identified and stored at -70°C using 20% glycerol.

As a result of strain identification, the genus Lactobacillus ( Lactobacillus ), the genus Petiococcus ( Pediococcus ), the genus Weissella , the genus Leuconostoc ( Leuconostoc ) and the genus Enterococcus ( Enterococcus ) 193 of 39 species (species) included canine strains were isolated.

Example 2: Preparation of lactic acid bacteria fermented product of peony extract

After crushing the dried peony root and powdering it, 10 times the weight of the powder was mixed with water and hot water extraction was performed at 121° C. for 15 minutes. The hot water extract was _{treated with sodium carbonate (Na 2} CO ₃ ) to pH 3.6 to 6.8. fit Each of the lactic acid bacteria isolated in Example 1 was cultured at 28 ° C. for 48 hours, then ^{inoculated into a hot water peony extract at a concentration of 6x10 9} CFU / ml, and then fermented at 28 ° C. for 48 hours, and the supernatant (lactic acid bacteria of the peony extract ferment) was obtained. After that, the obtained peony extract and each lactic acid bacterium fermented product of the peony extract were concentrated under reduced pressure and powdered.

Example 3: Identification of lactic acid bacteria strains having paoniflorin metabolism

Since paeoniflorin, which is an extract of peony or one of the indicator components of the peony, is known to have metabolic syndrome-related activity, but it was determined that the activity to treat the metabolic syndrome was not sufficient, the present inventors determined the bioconversion activity of the indicator component in the peony. We tried to search for strains with

To search for these strains, a peony extract was prepared by adding 10 times (w/v) water to 1 g of a dry weight of peony and extracting it at 121 degrees for 15 minutes, and each strain isolated in Example 1 was treated with 2 x 10 ⁹ CFU /ml concentration was inoculated and cultured in a culture room at 28°C for 48 hours. After that, the culture (supernatant) was extracted with 50% ethanol, and the chromatogram change was analyzed under UV 235nm, 15-100% methanol gradient conditions through HPLC using TSK-GEL ODS-100V 5㎛ column (4.6mm x 15cm). (Fig. 1).

As a result, it was confirmed that the peak of the peony extract fermented with LN180020 of the genus Leuconostok among the isolated strains decreased at about 15 minutes, specifically expected to be paoniflorin, and the peak of the new material increased at 14.2 minutes (Fig. One).

Example 4: Leukonostok genus ( Leuconostoc sp.) phylogenetic analysis of LN180020 lactic acid bacteria

For phylogenetic analysis of Leuconostoc sp. LN180020 (KCTC 13719BP), genomic DNA of Leuconostoc sp. LN180020 was extracted. The 16S rRNA gene was amplified using a primer set (Table 1), and the base sequence of 16S rRNA (SEQ ID NO: 1) was confirmed and then compared with the known sequence of the leukonostok genus.

Primer sequence information Primer name Base sequence (5'→3') (SEQ ID NO:) 27F-1492R_F AGAGTTTGATCCTGGCTCAG (2) 27F-1492R_R CGGTTACCTTGTTACGACTT (3)

The 16S rRNA nucleotide sequence of LN180020 in the leuconostock genus was confirmed through BigDye (R) Terminator v3.1 Cycle Sequencing Kits (Applied Biosystems), and the previously known leuconostock genus sequence is in the NCBI database (https://www. ncbi.nlm.nih.gov/), EzTaxon-e server (https://www.eztaxon-e.ezcloud.net/) and LPSN (https://www.bacterio.net/nocardioides.html) databases. and sequence comparison analysis was performed using BioEdit v7.2.6.1, MEGA7 program.

As a result, the leuconostok genus LN180020 showed the highest 16S rRNA sequence homology with 99.86% with the leukonostok citreum ATCC 49370T, followed by the leuconostock holzapfelii ( L. holzapfelii ) BFE 7000 ^T and 99.78%; Leukono stock lactis ( L. lactis ) JCM 6123 ^T and leukonostok palmae ( L. palmae ) TMW 2.694 ^T and 98.92%; Leukonostok Kimchi ( L. kimchii ) IMSNU 11154 ^T and 98.49%; Leukonostok Gelidum subsp. L. gelidum subsp. gasicomitatum LMG 18811 ^T and 98.14%; Leukonostok mecenteroides subsp. Mesenteroides ( L. mesenteroides subsp. mesenteroides ) ATCC 8293 ^T and 97.85%; Leukonostok pseudomesenteroides (L. pseudomesenteroides) NRIC 1777 ^T and 97.71%; And leukotostok rapi ( L. rapi ) LMG 27676 ^T and 97.63% of 16s rRNA sequence homology was shown, through which kimchi-derived leukonostok genus LN180020 is phylogenetically closest to leukonostok citreum ATCC 49370T. was found to be (FIG. 2).

Example 5: Bioconversion of Paoniflorin in Peony Extract

Peony extract was prepared by adding 10 times (w/v) water to 1 g of peony dry weight and extracting at 121 degrees for 15 minutes, Leuconostoc sp. LN180020 (Accession No.: KCTC 13719BP) 2 x It ^{was inoculated at a concentration of 10 9} CFU/ml and cultured for 48 hours in a culture room at 28°C. After that, the same amount of ethanol was added to the culture (supernatant) for extraction, followed by HPLC (with TSK-GEL ODS-100V 5 μm column (4.6 mm x 15 cm), 25°C, UV 230 nm, 15-100% methanol gradient conditions). High-performance liquid chromatography) was performed to analyze the presence or absence of bioconversion. Liquid chromatography mass spectrometry (LC-MS) analysis for identifying the bioconverted material was performed by dissolving the supernatant in methanol and using the QTrap 3200 connected with Turbolon Spray source (AB SCIEX, Singapore) (Luna C18 (2), 100 x 2.0 mm, 3 μm; guard cartridge system, security guard #KJ 0-4282; Phenomenex, USA). The column was maintained at 25°C with 100% methanol (A) and HPLC water (B) containing 0.04% trifluoroacetic acid at a flow rate of 1.0 mL/min, and the sample was sampled at 15% for the first 5 minutes. After flowing methanol, the methanol concentration was sequentially increased from 15% to 100% from 5 minutes to 20 minutes, maintained at 100% methanol for 20 minutes to 25 minutes, and from 100% to 15% from 25 minutes to 35 minutes The methanol concentration was decreased until , and after that, it was measured while maintaining 15% methanol for 5 minutes (Table 2).

HPLC conditions HPLC retention time (min) A; MeOH (%) B; Water (%) 0-5 15 85 5-20 15-100 85-0 20-25 100 0 25-35 100-15 0-85 35-40 15 85

As a result of performing HPLC to analyze the component converted by LN180020 in the leuconostok genus, the indicator substance decreased near the retention time of the fermented peony extract at about 15 minutes (FIG. 1), and corresponds to a new useful component at about 14.2 minutes It was confirmed that the peak to be increased. In addition, in order to confirm the reduced indicator substances and new useful components, each corresponding peak was analyzed by thin layer chromatography using silica gel 60G F254 glass plate in the developing solvent methanol:chloroform (1.5:5). As a result of separation and purification and LC-MS analysis, it was found that the indicator material at about 15 minutes was paeoniflorin with a molecular weight of 480.3, and after conversion at about 14.2 minutes, the new useful component material had a molecular weight of 642.3. was confirmed (FIG. 3).

The structure of the new useful ingredient in which the paoniflorin was converted was identified by Proton, Carbon, TOCSY, HSQC, and HMBC NMR, and the results are shown in FIG. 4 . As a result of structural identification, the material produced by bioconversion of paoniflorin was identified as β-gentiobiosyl paeoniflorin ( FIG. 4 ).

Example 6: Confirmation of cytotoxic, anti-diabetic and anti-obesity effects of fermented products of Leuconostoc sp. LN180020 of peony extract

6-1: Confirmation of cytotoxicity

In order to confirm the cytotoxicity of LN180020 fermented products of Leukonostok of peony extract, LN180020 fermented products of Leukonostok of peony extract were treated at different concentrations (3 and 30 μg/ml) in mouse L6 myotube cells and 24 After an hour, the MTT assay was performed.

As a result, it was confirmed that there was no significant effect on cell proliferation in the untreated control group, the non-fermented peony extract treated group, and the LN180020 fermented product of the Leukonostok genus of the peony extract in the mouse L6 myotube cells. It was found that the fermented product of Leuconostoc sp. LN180020 (KCTC 13719BP) has no cytotoxicity (FIG. 5).

6-2. Check the antidiabetic effect

In order to confirm the antidiabetic effect of the LN180020 fermented product in Leuconostok of the peony extract, mouse L6 myoblasts ^{were dispensed in a 24-well culture vessel at a cell count of 5x10 5} cells/ml, and then once every 2 days in a differentiation medium ( Incubated with low glucose (5 mM) DMEM, 2% Fetal Bovine Serum, 1% penicillin/streptomyc). On the 7th day of culture, pre-treated LN180020 fermented products in Leukonostok of peony extract by concentration (3 and 30 μg/ml), reacted for 30 minutes, and treated with [ ¹⁴ C]2-Deoxy-D-glucose for 5 minutes reacted. ^{After the reaction was completed, [ 14} C]2-deoxy-D-glucose-6-phosphate, which was absorbed into mouse L6 myoblasts and transformed, was measured with a radiometer to measure glucose uptake. As a positive control, 100 nM concentration of insulin (porcine insulin) was used.

As a result, in the group treated with the LN180020 fermented product of Leuconostok of the peony extract, the [ ¹⁴ C]2-deoxy-D-glucose-6-phosphate content in L6 myoblasts was about 55% to 80% compared to the untreated control group increased, and it was confirmed that it increased by about 25 to 35% compared to the non-fermented peony extract treatment group. This indicates that when the peony extract is fermented using Leukonostok genus LN180020, the antidiabetic activity can be increased by promoting glucose absorption into muscle cells compared to the non-fermented peony extract. Moreover, it can be seen that 100 nM of porcine insulin used as a positive control has an antidiabetic effect almost equivalent to 10 μM (3.57 μg/ml) of rosiglitazone, which is actually used as a diabetes treatment, but at a low concentration (3 μg/ml) ^[14 C]2-deoxy-D-glucose-6-phosphate content of the group treated with LN180020 fermented leuconostok of the peony extract of ㎖) was similar to that of the insulin 100 nM treatment group, and the ^{) [14} C]2-deoxy-D-glucose-6-phosphate content of the Leukonostok genus LN180020 fermented group of the peony extract was significantly increased compared to the insulin 100 nM treatment group (FIG. 6) .

On the other hand, to evaluate the antidiabetic effect of other lactic acid bacteria fermented products of peony extract, Lactobacillus plantarum subsp. As a result of evaluating the antidiabetic effect of the peony extract fermented with Plantarum LN180155 (Df2) or Lactobacillus paraplantarum (Df3) in the same manner as the above experimental method, [ ¹⁴ C]2-deoxy- in L6 myoblasts It was confirmed that the D-glucose-6-phosphate content was almost the same as that of the untreated control group. However, in the group treated with the peony extract fermented with Lactobacillus paraplantarum (Df3) at a concentration of 100 μg/ml, [ ¹⁴ C]2-deoxy-D-glucose-6-phosphate content in L6 myoblasts was untreated. It increased slightly compared to the control group, but there was no significant difference (FIG. 7).

As a result, when fermenting a peony extract using Leuconostoc sp. LN180020 (KCTC 13719BP), the activity of promoting glucose absorption into muscle cells compared to unfermented peony extract and peony extract fermented with other lactic acid bacteria It was judged to be an excellent antidiabetic increase. This effect appears to have increased antidiabetic activity by converting Paoniflorin into a new material by lactic acid bacteria of the Leuconostoc sp. LN180020 (KCTC 13719BP). was considered to be effective in preventing or improving

6-3: Confirmation of anti-obesity effect

The lipase inhibitory activity was evaluated to confirm the anti-obesity effect of the LN180020 fermented product of Leuconostok of the peony extract.

Lipase inhibitory activity was measured as follows. After dissolving 5mg/ml of lipase in 150ul of the reaction solution (100mM Tris-Hcl, pH 8.2), each experimental sample (LN180020 fermented product in peony extract leuconostok, peony extract and Orlistat) was mixed with a solution in which 100uM was dissolved. , reacted at 37 °C for 5 min. After that, 30ul of a solution of the substrate p-nitrophenyl palmitate (PNPP) or 4-nitrophenyl dodecanoate (PNPD) dissolved in 0.1% (W/V) in 5 mM sodium acetate (pH 5.0) solution was added, followed by reaction at 37°C for 12 hours. The results were measured with a spectrophotometer at a wavelength of 409 nm. Orlistat was used as a control, and as experimental samples, peony extract and LN180020 fermented peony extract leuconostok were used, respectively.

As a result, as shown in FIG. 8, Orlistat used as a positive control for lipase inhibitory activity showed inhibitory activity of 61.7% and 50.4%, respectively, on PNPP and PNPD substrates, and LN180020 fermented product of Leukonostok genus of peony extract In the treatment group, the inhibitory activity was 53.5% and 38.4%, respectively. On the other hand, the non-fermented extract peony extract showed 40.1% and 21.8% inhibitory activity against PNPP and PNPD substrates, respectively, so that the lipase activity inhibitory effect was significantly increased in the peony extract after fermentation of LN180020 in Leukonostok. was confirmed.

Example 7: Confirmation of efficacy in improving obesity, diabetes, dyslipidemia and fatty liver of fermented products of Leuconostoc sp. LN180020 of peony extract in animal models administered with high-fat diet

C57BL/6, 8-week-old, male mice were divided into 10 mice in each group as follows to form an experimental group, and a diet and each test substance were administered for a total of 6 weeks:

1) Normal diet

2) High-fat diet

3) High fat diet + metformin (Met) 100mg/kg/day administration group

4) High-fat diet + peony hot water extract 1000mg/kg/day administration group

5) LN180020 fermented extract in Leukonostok of high-fat Shii + peony hot water extract 1000mg/kg/day administration group

After the end of the experiment, blood was collected from each experimental group, and body weight, HbA1c, fasting blood sugar, HOMA-IR, total blood cholesterol (TC), triglyceride (TG), LDL, HDL, NEFA (non-esterified fatty acids), AST and ALT was measured.

The results for this are shown in FIGS. 9A to 9D and FIG. 10 .

As can be seen in FIGS. 9A to 9D and 10 , in the group administered with LN180020 fermented product of Leukonostok of the peony extract, body weight, HbA1c, fasting blood sugar, HOMA-IR, TC, TG, LDL, AST and ALT levels were notified. The mode was significantly reduced compared to the administration group, and it was confirmed that the HDL level significantly increased. The effect of the LN180020 fermented product of Leuconostok genus of the peony extract was found to be significantly higher than that of the peony hot water extract.

Example 8: Confirmation of the efficacy of β-gentiobiosyl paoniflorin in improving obesity, diabetes, dyslipidemia and fatty liver in an animal model administered with a high-fat diet

C57BL/6, 8-week-old, male mice were divided into 10 mice in each group as follows to form an experimental group, and a diet and each test substance were administered for a total of 6 weeks:

1) Normal diet

2) High-fat diet

3) High fat diet + metformin (Met) 100mg/kg/day administration group

4) High-fat diet + Paoniflorin (PF) 100mg/kg/day administration group

5) High-fat diet + β-gentiobiosil paoniflorin (βGPF) 100mg/kg/day administration group

After the end of the experiment, blood was collected from each experimental group, and body weight, HbA1c, fasting blood sugar, HOMA-IR, total blood cholesterol (TC), triglyceride (TG), LDL, HDL, NEFA (non-esterified fatty acids), AST and ALT was measured.

The results are shown in FIGS. 11A to 11D and 12 .

As can be seen in FIGS. 11A to 11D and 12 , body weight, HbA1c, fasting blood glucose, HOMA-IR, TC, TG, LDL, AST and ALT levels were higher in the β-gentiobiosil paoniflorin administration group. It was significantly reduced compared to this administration group, and it was confirmed that the HDL level was significantly increased. It was confirmed that the effect of the β-gentiobiosil paoniflorin administration group was significantly higher than that of paoniflorin.

Formulation Example 1: Preparation of a pharmaceutical formulation

1. Preparation of powder

2 g of fermented product of peony extract according to the present invention

1 g lactose

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

2. Preparation of tablets

100 mg of fermented product of peony extract according to the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional manufacturing method of tablets.

3. Preparation of capsules

100 mg of fermented product of peony extract according to the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

4. Preparation of pills

1 g of fermented product of peony extract according to the present invention

1.5 g lactose

1 g of glycerin

0.5 g of xylitol

After mixing the above components, it was prepared so as to be 4 g per ring according to a conventional method.

5. Preparation of granules

150 mg of fermented product of peony extract according to the present invention

Soybean extract 50 mg

glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, and then filled in a bag.

The fermented peony extract lactic acid bacteria or β-gentiobiosil paoniflorin of the present invention has very excellent lipase inhibitory activity, which promotes digestion and absorption by decomposing lipids, and promotes glucose absorption into muscle cells, It can be usefully used for the improvement of metabolic syndrome or the development of food and medicine for treatment, so it has very high industrial applicability.

[Accession number]

Name of deposit institution: Korea Research Institute of Bioscience and Biotechnology

Accession number: KCTC13719BP

Deposit date: 20181120

Address of the depository institution: (56212) Korea Research Institute of Bioscience and Biotechnology (KRIBB), 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do, Republic of Korea (56212)

Claims (12)

Peony ( Paeonia lactiflora ) Lactobacillus fermented extract or β-gentiobiosyl paeoniflorin (β-gentiobiosyl paeoniflorin) or a pharmaceutical composition for preventing or treating metabolic syndrome comprising a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition according to claim 1, wherein the metabolic syndrome is at least one selected from the group consisting of diabetes, obesity, fatty liver, dyslipidemia, arteriosclerosis, insulin resistance syndrome, and complications thereof. The pharmaceutical composition according to claim 2, wherein the fatty liver is at least one selected from the group consisting of non-alcoholic fatty liver, alcoholic fatty liver, and non-alcoholic steatohepatitis. The pharmaceutical composition according to claim 1, wherein the lactic acid bacteria is Leuconostoc citreum. The pharmaceutical composition according to claim 1, wherein the lactic acid bacteria is a Leuconostoc sp. LN180020 with accession number KCTC 13719BP. The pharmaceutical composition according to claim 1, wherein the composition inhibits the activity of lipase. The pharmaceutical composition according to claim 1, wherein the composition increases glucose uptake into muscle cells. The pharmaceutical composition according to claim 1, wherein the peony extract is extracted using water, a C1 to C6 lower alcohol, an organic solvent, or a mixture thereof. According to claim 1, wherein the β- gentiobiosyl paoniflorin is Leuconostoc sp. LN180020 strain (Accession No.: KCTC13719BP) characterized in that produced by bioconversion of paoniflorin pharmaceutical composition. Peony ( Paeonia lactiflora ) A food composition for preventing or improving metabolic syndrome, comprising fermented lactic acid bacteria of an extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof as an active ingredient. Use of lactic acid bacteria fermented product or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof of peony ( Paeonia lactiflora ) extract for manufacturing a preparation for the treatment of metabolic syndrome. Peony ( Paeonia lactiflora ) Administering an effective amount of a formulation containing a lactic acid bacterium extract or β-gentiobiosyl paeoniflorin or a pharmaceutically acceptable salt thereof to an individual in need thereof A method of treating metabolic syndrome, comprising.

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