KR20100070664A - Composition for preventing or treating of obesity, hyperlipidemia, fatty liver or diabetes comprising indole-3-carbinol derivatives as active ingredients - Google Patents

Abstract

PURPOSE: A composition containing indole-3-carbinol derivative is provided to reduce hepatic tissue neutral lipid and plasma triglyceride. CONSTITUTION: A composition for preventing or treating Obesity, hyperlipidemia, or diabetes contains indole-3 carbinol derivatives of chemical formula 1, 2, 3, 4, 5 or 6 as an active ingredient. The indole-3-carbinol derivatives are contained in extracts or fractions of Brassicaceae. The Brassicaceae is Brassica oleracea, Raphanus sativus, Brassica rapa, Armoracia rusticana, or Arabidopsis thaliana.

Description

Composition for Preventing or Treating of Obesity, Hyperlipidemia, Fatty Liver or Diabetes Comprising Indole-3-carbinol Derivatives as Active including indole-3-carbinol derivatives as an active ingredient Ingredients}

The present invention relates to a composition for the prevention or treatment of obesity, hyperlipidemia, fatty liver or diabetes, including cruciferous plant extract or indole-3-carbinol derivatives.

Due to the change in living environment, the visceral fat type obesity of modern people has increased, and the incidence of metabolic syndrome accompanying diabetes, hypertension, lipid metabolism abnormality, insulin resistance, etc. is increasing rapidly. These diseases increase the risk of mutual development and are common diseases associated with multiple metabolic changes such as aging, stress, and immune dysfunction.

According to the 2005 National Health and Nutrition Survey, 32% of Korean adults aged 20 and over were obese (35.2% of adult men and 28.3% of women). The incidence of childhood obesity in Korea has also increased rapidly.In 2005, 11.3% of elementary school students, 10.7% of middle school students, and 16% of high school students were classified as obese (BMI ≥ 25 kg / m ² ), and overweight (BMI ≥ 23 kg / m ^2). Or 17% of obese adolescents had metabolic syndrome.

This increase in overweight and obesity leads to an increase in the prevalence of chronic diseases, such as high blood pressure (30.2% male, 25.6% male), diabetes (9.0% male, 7.2% female), and high cholesterol in Koreans aged 30 and over in 2005. The prevalence of bloodemia (7.5% male and 8.8% female) is very high compared to other countries. In 2005, the mortality rate from diabetes was 35.5 per 100,000 people in Korea, more than 3-7 times higher than in Japan (5.9), the UK (7.5), or Germany (16.6). The increase in the number of diabetic patients in Korea is expected to increase to 3,000,000 and to 5,450,000 by 2030, which means that 10% of Koreans will become diabetic.

According to the statistics of the Korea Institute of Health and Social Affairs, the socioeconomic losses from obesity and obesity complications in 2006 are estimated to total KRW 2.1 trillion, including indirect expenses such as medical expenses and income loss from treatment. Accordingly, the government sets the main goal of national health promotion in 2010 to reduce adult obesity rate to less than 20% and adolescent obesity rate to less than 15%, and precisely define obesity and metabolic diseases as an implementation strategy to achieve the goal. And the method of measurement was decided.

Efficacy products are imported and sold in Korea in a large amount as a major obesity treatment. Among them, anti-obesity products such as xenical, reductyl, and excori are widely known. Xenical, which is an orlistat-based ingredient, is the world's first anti-obesity agent that inhibits the action of lipase to inhibit fat absorption, decreases total cholesterol and LDL cholesterol levels, improves blood sugar and lowers blood pressure. see. Reductil, the main ingredient of sibutramine (sibutramine), was approved by the FDA in 1997 and is sold in more than 30 countries around the world.It maintains high levels of serotonin and noradrenaline in the sympathetic nervous system, inducing sympathetic nervousness, appetite and satiety There is this. Lastly, Exorix is a non-prescription semi-finished product imported from France, increases heat generation, increases the body's basic metabolism, inhibits lipase activity, reduces intestinal fat absorption by 30%, and increases the production of noradrenaline. There is an action to increase.

Obesity can be the best treatment effect in combination with diet therapy, exercise therapy and behavioral therapy, but this method takes a lot of time and effort and difficult to implement, so many obesity treatments or diet products are used. However, Olistat, which is currently used as a treatment for obesity, has side effects such as fatty stool, intestinal gas generation, and bloating. Sibutramine is known to have side effects such as headache, dry mouth, anorexia, insomnia, and constipation. Orlistat also inhibits the absorption of vitamin E and vitamin D, while phentermine and sibutramine have side effects that lead to increased heart rate, palpitations, or dizziness.

As Western medicine shows limitations in overcoming side effects and chronic diseases of synthetic drugs, the value of new natural products is emerging. Accordingly, the present inventors found indole-3-carbinol, which is a phytochemical compound contained in Brassica family plants, such as broccoli and cabbage, in the process of searching for an active substance that suppresses obesity from native plants. indole-3-carbinol).

Indole-3-carbinol is a chemical mixture extracted from brassica plants such as broccoli and cabbage, and has a structural formula of C ₉ H ₉ NO and a molecular weight of 147.17. Indole-3-carbinol is an adjunct of the indole group which is a grayish white solid and is produced by digestion or cooking processes such as cutting, grinding, chewing, and enzymes in vivo. It is made from the precursor indole-3-glucosinolate by the enzyme myrosinase.

The main physiological functions of indole-3-carbinol known to date are antioxidant and anticancer activity. Indole-3-carbinol prevents cell damage by free radicals, and in particular, maintains health by inhibiting cell damage of the breast, prostate or reproductive organs. Treatment of prostate cancer cells with indole-3-carbinol at a concentration of 25-100 μM resulted in blocking the cell cycle and inhibiting cancer cell proliferation by stopping the G1 checkpoint. Indole-3-carbinol treatment increases the expression of tumor suppressor genes p21 and p27 in cancer cells, while reducing the expression of NF-κB. In addition, the administration of indole-3-carbinol in 18 patients with papilloma had stopped papilloma growth so that 12 patients would not have to undergo surgery. In the functional food market, indole-3-carbinol is classified as a dietary supplement and is sold as an antioxidant.

U.S. Pat. U.S. Pat.

US Pat. No. 6,871,131 discloses a composition for treating hyperlipidemia comprising phenylacetylglutamine, phenylacetylisoglutamine phenylacetic acid. US Pat. No. 6942967 discloses apobec-1 protein as a therapeutic target for atherosclerosis, hyperlipidemia, obesity and diabetes.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors endeavored to develop natural substances having activities such as anti-obesity, anti-hyperlipidemia and / or anti-diabetic. As a result, the present invention was completed by confirming that indole-3-carbinol contained in the extract of Cruciferae / Brassicaceae plant has the above activity.

Accordingly, an object of the present invention is to provide a composition for the prevention or treatment of obesity, hyperlipidemia, fatty liver or diabetes comprising indole-3-carbinol derivatives as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to an aspect of the present invention, there is provided a composition for preventing or treating obesity, hyperlipidemia, fatty liver or diabetes, including indole-3-carbinol as an active ingredient. Indole-3-carbinol derivatives have the structure of Formula 1:

Formula 1

The present inventors endeavored to develop natural substances having activities such as anti-obesity, anti-hyperlipidemia and / or anti-diabetic. As a result, it was found that indole-3-carbinol contained in cruciferous plants has the above activity.

The indole-3-carbinol exhibits prophylactic or therapeutic activity for hyperlipidemia, fatty liver or obesity. As demonstrated in the examples below, indole-3-carbinol inhibits adipocyte differentiation, decreases body fat mass, decreases visceral fat mass, decreases total cholesterol concentration, decreases plasma triglycerides and hepatic triglycerides Efficacy in relieving the obesity phenomenon induced by diet significantly. Indole-3-carbinol also exerts a significant decrease in fasting blood glucose and blood insulin concentrations, thereby improving the type 2 diabetes or insulin resistance and the effects of improving metabolic inflammatory responses closely related thereto.

According to another aspect of the present invention, indole-3-carbinol comprises chemical derivatives and biologically converted derivatives having the above prophylactic or therapeutic activity for hyperlipidemia, fatty liver or obesity. For example, indole-3-carbinol is extremely unstable in acidic solutions and changes to other derivatives by biological activity when exposed to gastric acid (de Kruif, CA, et al. (1991) Chem.-Biol.Interact. 80: 303-315.), These derivatives have been found in the intestinal components of rats fed either a basic diet or indole-3-carbinol (Grose, KR, and Bjeldanes, LF (1992) Chem. Res. Toxicol 5: Bradfield, CA and Bjeldanes, LF (1987). J. Toxicol. Environ. Health 21: 311-323 .; Bjeldanes, LF, et al. (1991). Proc. Natl. Acad. Sci USA 88 Kwon, C.-S., et al. (1994) J. Agr. Food Chem. 42: 2536-2540. Indole-3-carbinol is prepared by the catalyzed acid catalysis under low pH conditions of the stomach and 3,3'-diindolylmethane (DIM) and indolo [3,2-b] carbazole ( converted to a dimer, a biologically active molecule such as indolo [3,2-b] carbazole (ICZ) (Bjeldanes, LF, et al. (1991). Proc. Natl. Acad. Sci USA 88: 9543-9547.). ICZ, on the other hand, may be produced from indole, tryptophan, etc. as a metabolite of gut bacteria (Kwon, C.-S., et al. (1994) J. Agr. Food Chem. 42: 2536-2540.).

In addition to the derivatives, a variety of indole-3-carbinol derivatives are produced through degradation processes such as cutting, grinding, or chewing of cruciferous plants, or reactions caused by cooking processes or enzymes in vivo, and typical 5,6,11,12 , 17,18-hexahydrocyclone [1,2-b: 4,5-b ': 7,8-b ”] triindole (5,6,11,12,17,18-hexahydrocyclon [1,2- b: 4, 5-b ': 7,8-b ”] triindole, CTr), 5,10-dihydroindeno [1,2-b] indole (5,10-Dihydroindeno [1,2-b] indole), 3- (Methoxymethyl) indole, etc. (Jin Seok Kang et al. (2000) Journal of Korean Association of Cancer Prevention. 5 (4): 199-208.) .

As described above, indole-3-carbinol that is unstable in aqueous solution forms derivatives by the formation of vinyly hemiamine (vinylogous hemiaminal), and easily dehydrates to become 3-methyleneindolenium cation (Grose , KR, and Bjeldanes, LF (1992) Chem. Res. Toxicol 5: 188-193 .; Bjeldanes, LF, et al. (1991) Proc. Natl. Acad. Sci USA 88: 9543-9547.). The cation can be directly reacted with various nucleophiles, including indole-3-carbinol, to synthesize or prepare a variety of derivatives having biologically equivalent or similar activity with indole-3-carbinol. Representative examples of the derivatives are shown in Table 1.

Representative chemical and biologically converted derivatives of indole-3-carbinol Name of indole-3-carbinol derivative Chemical formula 3,3'-diindoleylmethane (DIM)

Indolo [3,2-b] carbazole (ICZ)

5,6,11,12,17,18-hexahydrocyclone [1,2-b: 4,5-b ': 7,8-b ”] triindole (CTr)

5,10-dihydroindeno [1,2-b] indole

3- (methoxymethyl) indole

According to a preferred embodiment of the present invention, the indole-3-carbinol or a derivative thereof is included in the extract or fraction of the cruciferous plant.

The cruciferous plant is an angiospermae called Brassicaaceae or Cruciferae, and the name Brassicaea is derived from the genus Brassica. Crucifera is an older name and has “cross-bearing” because four petals are crossed to imply a cross. The cruciferous plant, which contains a large amount of the phytochemical compound indole-3-carbinol, is known to contain more than about 3,700 species of the genus 330, according to the Royal Botanic Gardens, Kew of England. Cruciferous plant used in the present invention is an indole-3-carbinol is not particularly limited including a play derivative, preferably a Brassica oleic la chair cabbage (cabbage), broccoli (broccoli) belonging to the (Brassica oleracea), E. coli flower (cauliflower), Brussels's infrastructure in Lodz (brussels sprouts), collard (collards), kale (kale), such as Brassica rapa turnip (turnip), cabbage (Chinese cabbage) belonging to (Brassica rapa), such as Brassica or crispus (Brassica napus), such as rapeseed (rapeseed) belongs to, Rafa Taunus Satie booth etc. Common free (radish) belongs to the (Raphanus sativus), scented shea ruseutikana (Armoracia rusticana) horseradish (horseradish), Marty climbed belonging to ( Stoke is a (stock), Talia and cis arabian doped (Arabidopsis thaliana) belonging to Matthiola), etc. (Hall, JC, KJ Sytsma and HH Iltis. 2002. phylogeny of Capparaceae and Brassicaceae based on chloroplast sequence data. American Jour nal of Botany 89: 1826-1842; Walter S. Judd, Christopher S. Campbell, Elizabeth A. Kellogg, Peter F. Stevens, Michael J. Donoghue: Plant Systematics: A Phylogenetic Approach , Sinauer Associates Inc. 2007; Stevens, PF (2001 onwards) Angiosperm Phylogeny Website.Version 7, May 2006; Strasburger, Noll, Schenck, Schimper: Lehrbuch der Botanik f Hochschulen. 4. Auflage, Gustav Fischer, Jena 1900, p. 459).

Cruciferous plant extracts comprising indole-3-carbinol derivatives can be obtained using conventional extraction solvents for cruciferous plants (preferably, the cruciferous plants), preferably (a) anhydrous having 1 to 4 carbon atoms. Or a hydrous lower alcohol (e.g. methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol and normal-butanol, etc.), (b) a mixed solvent of the lower alcohol with water, (c) acetone, (d) Ethyl acetate, (e) chloroform, (f) 1,3-butylene glycol, (g) hexane, (h) diethyl ether, (i) butyl acetate or (j) water can be obtained as an extraction solvent.

Cruciferous plant fractions comprising indole-3-carbinol derivatives refer to more isolated / purified forms obtained by additional separation / purification of cruciferous plant extracts. For example, fractions obtained by passing cruciferous plant extracts through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods are also included in the cruciferous plant fraction.

In addition, the indole-3-carbinol derivative may be chemically synthesized.

According to a preferred embodiment of the present invention, indole-3-carbinol derivatives include indole-3-carbinol derivatives prepared synthetically in addition to those isolated from the cruciferous plants.

According to another aspect of the invention, the present invention provides a pharmaceutical composition or food composition for the prevention or treatment of obesity, hyperlipidemia, fatty liver or diabetes, including indole-3-carbinol derivatives.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and preferably applied by oral administration.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. Preferred dosages of the pharmaceutical compositions of the invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

When the composition of the present invention is prepared as a food composition, it includes not only indole-3-carbinol derivatives as active ingredients, but also components commonly added in the manufacture of food, and include, for example, proteins, carbohydrates, fats, nutrients, Seasonings and flavoring agents. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents [tautin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared with a drink, in addition to the indole-3-carbinol derivative of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. May be additionally included.

The composition of the present invention comprising an indole-3-carbinol derivative as an active ingredient, inhibits adipocyte differentiation, decreases body fat amount, decreases visceral fat amount, decreases total cholesterol concentration, decreases plasma triglycerides and hepatic triglycerides, It results in a decrease in fasting blood sugar and a decrease in blood insulin concentration, ultimately exhibiting prophylactic or therapeutic activity of obesity, hyperlipidemia, fatty liver or diabetes.

As used herein, the term "hyperlipidemia" refers to a disease caused by a large amount of fat in the blood due to poor metabolism of triglycerides and cholesterol. More specifically, hyperlipidemia refers to high cholesterol hyperlipidemia, which occurs frequently with increased lipid components such as triglycerides, LDL cholesterol, phospholipids, and free fatty acids in the blood.

As used herein, the term “fatty liver” refers to a condition in which fat accumulates in hepatic cells due to adipose metabolism disorder of the liver, which causes various diseases such as angina, myocardial infarction, stroke, arteriosclerosis, fatty liver and pancreatitis. .

As used herein, the term “diabetes” means a chronic disease characterized by a relative or absolute lack of insulin resulting in glucose-intolerance. The term diabetes includes all kinds of diabetes and includes, for example, type 1 diabetes, type 2 diabetes and hereditary diabetes. Type 1 diabetes is insulin dependent diabetes mellitus, mainly caused by the destruction of β-cells. Type 2 diabetes is insulin-independent diabetes, caused by insufficient insulin secretion after meals or by insulin resistance.

According to a preferred embodiment of the present invention, the composition of the present invention is applied to the prevention or treatment of type 2 diabetes, more preferably type 2 diabetes by insulin resistance.

The features and advantages of the present invention are summarized as follows.

(Iii) The present invention provides a composition for the prevention or treatment of obesity, hyperlipidemia, fatty liver or diabetes, comprising an indole-3-carbinol derivative as an active ingredient.

(Ii) The composition of the present invention can be separated using a cruciferous plant, and can also be synthesized through a chemical method.

(Iii) The composition of the present invention comprising cruciferous plant extract or indole-3-carbinol derivatives as an active ingredient inhibits adipocyte differentiation, decreases body fat amount, decreases visceral fat amount, decreases total cholesterol concentration, plasma triglycerides and It results in a decrease in hepatic triglycerides, a decrease in fasting blood glucose and a decrease in blood insulin levels, ultimately exhibiting prophylactic or therapeutic activity in obesity, hyperlipidemia, fatty liver or diabetes.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Example 1: Inhibitory effect of indole-3-carbinol on adipocyte differentiation using mouse adipocyte line (3T3-L1)

Cell culture and oil red O staining

Mouse fat cell lines (3T3-L1, ATCC, American Type Culture Collection, Manassas, VA, USA) were used to evaluate the effects of indole-3-carbinol (Sigma, USA) on the differentiation and growth of adipocytes. Adipocytes, 3T3-L1 cells, were dispensed in 12-well plates and added Dulbecco-modified Eagle with 1% penicillin-streptomycin, 1% non-essential amino acids, and 10% fetal bovine serum (FBS). medium) was used to incubate in a confluent state at 37 ℃, 5% CO ₂ incubator. Confluent and grown 3T3-L1 cells were cultured for two days in a medium containing MDI (0.5 mM isobutyl-methylxanthine, 1 μM dexamethasone and 1 μg / ml insulin) and differentiated. After making adipocytes (differentiation adipocytes), the cells were differentiated into mature adipocytes while incubated for two more days in a DMEM medium containing 1 μg / ml insulin. Thereafter, the cells were incubated for 10 days while replacing the DMEM medium every two days to form fully differentiated adipocytes.

Indole-3-carbinol was treated in culture at concentrations of 0.1, 1, 10, 50 and 100 μM at two-day intervals from the first day of differentiation by adding MDI to 3T3-L1 cells. Indole-3-carbinol was used by dissolving in DMSO, and negative control group containing only DMSO was included in the experiment. After incubation for a total of 14 days, the culture solution was removed at the time of differentiation was completed, and stained fat cells contained in the differentiated adipocytes. To do this, wash the cells twice with PBS (phosphate buffered saline), fix the cells for 1 hour with 10% BNF (Buffered Neutral Formalin), wash once again with PBS, and then oil red specifically to stain red fat. 1 ml of O dye was added to a 12-well plate to stain the fat globules for 1 hour and washed twice with distilled water.

In order to measure the concentration of triglycerides contained in the differentiated 3T3-L1 cells, stained fat globules were dissolved in 1 ml of isobutanol, and the O.D values were measured at 600 nm.

Indole-3-carbinol Inhibits Adipose Cell Differentiation

Treatment with 3T3-L1 cells with indole-3-carbinol as shown in panel A of FIG. 1 significantly reduced the differentiation of adipocytes in a concentration dependent manner. The amount of intracellular fat stained with oil red O was quantified by spectrophotometry, which also reduced the O.D value in a concentration dependent manner (FIG. 1, Panel B).

Example 2: Body weight and visceral fat reduction effect of indole-3-carbinol using a mouse

Experimental diet preparation and breeding of laboratory animals

The obesity induction diet used in this experiment was a high fat diet (HFD: 40% fat calorie, 17 g Laad + 3% corn oil / 100 g diet) and the diet containing indole-3-carbinol was HFD The composition is the same as indole-3-carbinol contained 0.1% level. Normal diet (ND) was prepared based on the composition of the AIN-76 rodent diet (American Institute of Nutrition, Report of the American Institute of Nutrition ad hoc committee on standards for nutritional studies. J. Nutr. 107: 1340-1348, 1977). (Table 2).

Experimental Formula ingredient Normal diet
(g / kg diet) Control diet
(g / kg diet) Indole-3-carbinol diet
(g / kg diet) Casein 200 200 200 DL-Methionine 3 3 3 Corn starch 149.99 110.96 109.96 Sucrose 500 370 370 cellulose 50 50 50 Corn oil 50 30 30 Laad - 170 170 Vitamin complex 10 12 12 Mineral complex 35 42 42 Choline Bitartrate 2 2 2 cholesterol - 10 10 tert-butyhydroquinone 0.01 0.04 0.04 Indole-3-carbinol - - 1.00 Total (g) 1,000 1,000 1,000 Fat (% calories) 11.5 39.0 39.0 Total calories, kJ / kg diet 16,439 19,315 19,315

Five-week-old male C57BL / 6J mice (Orient, Korea) were adapted to the laboratory environment for 1 week with solid feed, and then randomly placed into the high fat diet control group and the experimental group according to the egg mass method, and reared for a total of 10 weeks. The diet was fed with water between 10 and 11 am daily, and dietary intake was measured daily and body weight was measured every three days. In order to prevent sudden weight change due to feed intake, body weight was measured 2 hours after removing the feed container, and the dietary efficiency was the total test period from the feeding date to the sacrifice day, and the cumulative weight gain during the test period was determined by the total diet. Calculated by dividing by intake. After fasting the test animals for 12 hours or more, blood, liver, and visceral fat tissue (diplordial fat, perirenal fat, mesenteric fat, and abdominal fat) were collected under anesthesia with diethyl ether to prepare 0.1 M phosphate buffer solution (pH). 7.4), and then weighed. Blood collected from the abdominal aorta was centrifuged at 1000 x g for 15 minutes to separate plasma.

Body weight and visceral fat weight

The final body weight and the weight gain for the 10 weeks after ingesting the experimental diet for 10 weeks, the final body weight was 36%, and cumulative in the group fed the indole-3-carbinol compared to the high-fat diet control group (HFD) Weight gain decreased 59% significantly (FIG. 2).

After eating the experimental diet for 10 weeks, the weight of epididymal fat, periphery fat, mesenteric fat, and posterior cavity fat were measured, and the epididymis converted to unit weight in the indole-3-carbinol group compared to the control group (HFD). Fat weight was 44%, peripheral fat weight 74%, mesenteric fat weight 57%, posterior cavity fat weight 46%, and total visceral fat weight 49% ( P <0.001). Therefore, it was confirmed that indole-3-carbinol had an excellent weight loss and visceral fat reduction effect (FIG. 3).

Example 3: Prevention and Treatment Efficacy of Indole-3-carbinol in Obese Hyperlipidemia, Fatty Liver and Type 2 Diabetes

Plasma total cholesterol, triglyceride and glucose concentration were measured twice each using a commercial clinical kit (Bio Clinical system), insulin concentration was measured by ELISA using a mouse insulin kit (Shibayaki, Japan). The lipid component of liver tissue was extracted according to the method of Folch et al. 1 mL of distilled water was added to 0.25 g of liver tissue and homogenized using a polytron homogenizer (IKA-WERKE GmbH & Co., Ultra-Turrax, Staufen, Germany). 5 mL of chloroform: methanol solution (2: 1, v / v) was added to the homogeneous solution, mixed well, and the lower layer was separated by centrifugation at 1000 xg for 10 minutes, and the chloroform: methanol solution (2: 1, v / v) 2 mL was added, and the same procedure was repeated to completely separate the liver lipid components. 3 mL of chloroform: methanol: 0.05% CaCl ₂ (3:48:47, v / v / v) solution was added to the lower layer solution, mixed for 1 minute, and centrifuged at 1000 xg for 10 minutes. After completely drying with nitrogen gas, the dried lipid was dissolved in 1 mL of methanol and used for lipid component analysis. Triglyceride concentrations in hepatic lipid extracts were measured using the same commercial lipid analysis kit (Bio Clinical system) used for the analysis of plasma.

Plasma lipid concentrations of mice fed the experimental diet shown in Table 2 for 10 weeks were 72% triglyceride and 54% total cholesterol in the indole-3-carbinol group compared to the high-fat diet control group (HFD). , LDL + VLDL cholesterol concentration was 63%, free fatty acid concentration was significantly reduced by 48% (Table 3).

Dietary obesity It is well known that animal type or human obesity is accompanied by type 2 diabetes, which increases blood insulin levels and increases fasting blood glucose. In addition, the term 'metaflammation' has recently appeared for inflammatory reactions caused by nutrient or metabolic oversupply, and obesity is interpreted as 'chronic and low-level inflammation'. There is an ongoing research into the relationship between obesity and the immune system. In other words, a variety of obesity complications (type 2 diabetes, insulin resistance, arteriosclerosis, cancer and asthma, etc.) is a new view that is caused by the interaction with the immune system in the course of obesity. For example, TLR4 (toll-like receptor 4), which is involved in the innate immune response, uses dietary fats (particularly saturated fatty acids) as ligands, acting as an important factor in the inflammatory response and insulin resistance pathways, while in the central nervous system It is also suggested to be involved in appetite control. Therefore, the concentration of free fatty acids in blood and tissues has recently been recognized as a major indicator of the development of insulin resistance and metabolic diseases caused by obesity.

In this study, ingestion of indole-3-carbinol for 10 weeks in mice fed high fat diet resulted in a 22% reduction in fasting blood glucose and a 36% significant decrease in blood insulin levels compared to the high fat diet control group. Insulin resistance index (IRI) was also significantly reduced by 50% (Table 3). Therefore, indole-3-carbinol has an effect of improving type 2 diabetes or insulin resistance, and is also expected to improve the metabolic inflammatory response closely related thereto.

Obesity-related Biochemical Indicators of Mouse Blood Ingested with Indole-3-carbinol Normal diet.
(ND) High Fat Diet Control Group
(HFD) Indole-3-carbinol group Triglycerides (mmol / L) 0.60 ± 0.09 1.41 ± 0.12 0.39 ± 0.09 ^* Total Cholesterol (mmol / L) 3.07 ± 0.09 6.20 ± 0.54 2.84 ± 0.19 ^* HDL cholesterol (mmol / L) 1.33 ± 0.09 2.33 ± 0.27 1.42 ± 0.08 ^* LDL + VLDL Cholesterol (mmol / L) 1.74 ± 0.06 3.87 ± 0.57 1.42 ± 0.22 ^* Free fatty acid (uEq / L) 357 ± 26 534 ± 8 ^* 280 ± 40 ^* Glucose (mmol / L) 6.16 ± 0.95 8.49 ± 0.53 6.63 ± 0.31 ^* Insulin (ng / mL) 0.51 ± 0.06 0.76 ± 0.10 0.48 ± 0.06 ^* IRI ^a 0.54 ± 0.07 1.10 ± 0.10 0.55 ± 0.08 ^*

^* Significantly different from the value for HFD group by Student's t-test at P <0.001.

^a Insulin resistance index (IRI) = 10 ^-3 pmol insulin x mmol glucose x L ^-2

The liver weight of the mice fed the experimental diet for 10 weeks was significantly reduced by 24% in the indole-3-carbinol group compared to the high-fat diet control group. The lipid concentrations of liver tissues were significantly lower in the indole-3-carbinol-treated group than in the high-fat diet control group (89%), cholesterol (87%), cholesterol (65%), and free fatty acid (65%). ). Therefore, it can be seen that indole-3-carbinol remarkably alleviates the fatty liver phenomenon in high fat diet-induced obesity and significantly improves obesity inflammation and insulin resistance in the liver tissue.

Obesity-related Biochemical Indicators of Mouse Liver Tissue Ingested with Indole-3-carbinol Normal diet.
(ND) High Fat Diet Control Group
(HFD) Indole-3-carbinol group Liver weight (g / 100 g body wt) 3.6 ± 0.1 5.5 ± 0.3 4.2 ± 0.3 ^* Triglyceride (μmol / g) 20.4 ± 0.30 35.1 ± 0.32 3.78 ± 0.20 ^* Cholesterol (μmol / g) 21.6 ± 0.58 74.8 ± 0.68 9.4 ± 0.10 ^* Free fatty acids (uEq / g) 8.56 ± 0.35 24.3 ± 0.22 ^* 8.47 ± 0.13 ^*

^* Significantly different from the value for HFD group by Student's t-test at P <0.001.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

1 is a diagram showing the effect of inhibiting the differentiation of indole-3-carbinol in 3T3-L1 cells.

2 is a diagram showing the body weight and weight gain of the mice ingested the experimental diet. The blue solid line represents the normal diet group (ND), the red solid line represents the high fat diet control group (HFD), and the light blue solid line represents the indole-3-carbinol administration group.

3 is a diagram showing the weight vs. epididymal fat, peripheral kidney fat, mesenteric fat, celiac fat and total visceral fat weight of mice fed the experimental diet. The blue bar graph shows the normal diet group (ND), the red bar graph shows the high-fat diet control group (HFD), and the light blue bar graph shows the indole-3-carbinol administration group.

Claims (5)

A composition for preventing or treating obesity, hyperlipidemia, fatty liver or diabetes, comprising an indole-3-carbinol derivative represented by any one of the following Chemical Formulas 1 to 6 as an active ingredient: Formula 1

Formula 2

Formula 3

Formula 4

Formula 5

Formula 6

The composition of claim 1, wherein the indole-3-carbinol derivative is included in an extract or fraction of a Cruciferae / Brassicaceae plant. The method of claim 2 wherein the cruciferous plant is Brassica oleic la chair (Brassica oleracea), Brassica rapa (Brassica rapa), Brassica or crispus (Brassica napus), Rapa Taunus Sati booth (Raphanus sativus), aromatic cyano ruseutikana ( Armoracia rusticana), Marty up (Matthiola) and arabinose-doped systems Italia or (Arabidopsis thaliana) with a composition being selected from the group consisting of. (a) a therapeutically effective amount of the composition of any one of claims 1 to 3; And (b) a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia, fatty liver or diabetes comprising a pharmaceutically acceptable carrier. (a) a food-effective amount of the composition of any one of claims 1 to 3; And (b) a food composition for preventing or improving obesity, hyperlipidemia, fatty liver or diabetes, which includes a food acceptable carrier.

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