KR101722297B1 - ISOLATION OF SINGLE COMPOUND FROM Albatrellus fletti ITS APPLICATION IN TREATING AND PREVENTING OBESITY - Google Patents

Abstract

(1)The present invention relates to a compound extracted from albatrolus pluti mushroom, and more particularly, to a compound for treating or preventing obesity, which is isolated from an alcoholic extract of albatrolus pluti mushroom which is effective for the treatment and prevention of obesity, .
The compound of the present invention is represented by the following formula (1).

(One)

Description

Technical Field [0001] The present invention relates to a compound for treating or preventing obesity, which is derived from albatrolus platelis.

The present invention relates to a compound extracted from albatrolus pluti mushroom, and more particularly, to a compound for treating or preventing obesity, which is isolated from an alcoholic extract of albatrolus pluti mushroom which is effective for the treatment and prevention of obesity, .

Obesity is a disease in which excess fat tissue accumulates in the body. Obesity is a major cause of various cancers such as cardiovascular disease, type 2 diabetes, colorectal cancer, rectal cancer, prostate cancer, breast cancer, and ovarian cancer. Obesity is mainly caused by excessive calorie meals and lack of exercise, resulting in the accumulation of surplus energy sources in the form of fat in the body. Treatment of obesity using drugs is attracting attention.

Drugs for treating obesity include direct drugs acting on the neurotransmitter or hormone system to give fullness, suppression of appetite, direct drugs, and indirect drugs acting on the digestive enzymes to inhibit the absorption of fat.

Xenical ^TM (orlistat) is a well-known indirect drug, which covalently binds to serine, the active site of triglyceride lipase (TGL), which inhibits the activity of TGL, do.

"Development of specific egg yolk antibodies to the pancreatic lipase domain for obesity inhibition" Kor. J. Microbiol. Biotechnol. Vol. 36, No. 4, 299-306 (2008) proposes a method of inhibiting the activity of pancreatic lipase by inhibiting the absorption of fat by using a specific egg yolk antibody to inhibit obesity.

The selection of the present inventors in PCT / KR2012 / 10558, PCT / KR2012 / 10602, and PCT / KR2012 / 10621 discloses that the extracts of Pseudomonas aeruginosa and Abacus japonicus mushroom inhibit the activity of pancreatic lipase, respectively.

However, the prior art fails to identify a compound exhibiting efficacy as an obesity therapeutic agent, and refers only to the efficacy of the extracts.

The object of the present invention is to provide a compound which exhibits almost no in vivo side effects, inhibits the activity of pancreatic lipase and exhibits the therapeutic or preventive effect of obesity caused by triglycerides, medicines containing the same, and a health functional food.

In order to accomplish the above object, the present invention provides a composition for treating or preventing obesity comprising an effective amount of a compound of the following formula (1) or an extract of albatrossus pleti mushroom containing the same.

(1)

(One)

The present invention also provides a medicament for the treatment or prevention of obesity comprising the compound of the above formula (1) or an extract of albatrolus pleti mushroom containing the compound as an active ingredient.

The present invention also provides a method of treating or preventing obesity comprising the step of administering to a animal an extract of Albatrilus pluti mushroom comprising the compound of formula (1) or the compound.

The present invention also provides a health functional food for improving or preventing obesity, which comprises the compound of the above formula (1) or the extract of Albatrilus pluti mushroom containing the compound as an active ingredient.

The present invention also relates to the use of the compound of formula (1) or the albatellus pleti mushroom containing the compound effectively inhibits the activity of pancreatic lipase, which is a triglyceride lipase (TGL) Can be used for prevention.

The present invention also relates to a method for extracting a composition for preventing or treating obesity comprising the compound of formula (1) or the compound of formula (1) from albatrolus pluti mushroom using an alcohol having 1 to 10 carbon atoms to provide. The alcohol is preferably a lower alcohol having 1 to 5 carbon atoms, and most preferably ethanol. The ethanol may be 90 to 99.99% (v / v) ethanol.

In the present invention, the alcohol extract of Albatrilus pluti mushroom containing the compound of formula (1) may be one prepared by a conventional method for producing plant extracts. Preferably, the albatolus pluti mushroom is dry-pulverized and then the residue is removed. Then, 0.1 to 20 g of the pulverized product is added per 100 ml of the alcohol having 1 to 10 carbon atoms, more preferably, 100 ml of the extraction solvent 1 to 5 g may be added to the solution. When the content of the albatrolus pleti mushroom pulverized product is too small as compared with the extraction solvent, the effect of albatrolus pleti mushroom is not sufficient and the effect is increased due to an increase in the content when the amount of the solvent is too large relative to the amount of the extraction solvent But it is not preferable from the viewpoint of productivity because the production cost is increased. At this time, the extraction conditions are not limited, but it is preferable to mix the albatrolus pluti mushroom with an alcohol having 1 to 10 carbon atoms as an extraction solvent, and then, at a temperature of 20 to 60 ° C for 12 to 36 hours, Can be extracted at a temperature of 30 to 40 占 폚 for 20 to 24 hours. Extraction at an excessively low temperature condition requires a long time for extracting an active ingredient, and extraction at an excessively high temperature may deteriorate the activity, which is undesirable. When the extraction time is too short, the concentration of the effective component to be extracted is low, and when the extraction is performed for an excessively long time, the concentration of the extraction effective ingredient increases with the increase of the extraction time. Then, the alcohol extract of Albatrolus pluti mushroom extracted by the above method is filtered through a filter cloth, and the filtrate is centrifuged to remove the precipitate, and then the filtrate can be concentrated or concentrated under reduced pressure, followed by lyophilization.

In the present invention, the compound of the above formula (1) can be separated from the alcohol extract of Albatrilus pluti mushroom by HPLC. In a preferred embodiment, the alcohol extract can be obtained by dissolving the alcohol extract in a solvent, collecting a range of fractions and then fractionating again with HPLC.

On the other hand, the content of the compound of formula (1) as an active ingredient in the composition for treating or preventing obesity in the above-described embodiment is not limited, but may be preferably 0.001 to 50% by weight. When the content of the compound of the formula (1), which is an effective ingredient, is less than 0.001% by weight, the digestion and absorption of the fat in the body is insignificant, while when it exceeds 50% by weight, It is not economical. More preferably, the content of the alcohol extract of albatolus plutium mushroom in the composition may be 0.01 to 50% by weight, and most preferably 0.1 to 30% by weight.

The present invention provides a medicament for treating or preventing obesity comprising the compound of formula (1) as an active ingredient according to an embodiment. Such medicines containing the compound of the above formula (1) as an active ingredient may further contain suitable carriers, excipients and diluents conventionally used in the production thereof.

Examples of carriers, excipients and diluents that can be included in the pharmaceutical agent containing the compound of the formula (1) of the present invention as an active ingredient include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, , Alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil have.

Each of the medicines containing the compound of formula (1) of the present invention as an active ingredient may be used as an oral preparation such as powders, granules, tablets, suspensions, emulsions or syrups according to conventional methods.

The oral formulations are meant to include solid and liquid formulations for oral administration and solid formulations for oral administration may include tablets, pills, powders, granules, capsules and the like, For example, starch, calcium carbonate, sucrose or lactose, gelatin and the like in combination with at least one excipient. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, have.

The preferred dosage of the composition containing the compound of formula (1) of the present invention varies depending on the condition and body weight, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. Preferably, the medicinal product containing the alcohol extract of albatrolus plutium mushroom of the present invention as an active ingredient is administered at a dose of 0.0001 to 100 mg / kg per day based on the amount of the alcohol extract of albatrolus pleti mushroom, It is preferable to administer it at 0.01 to 10 mg / kg. The administration may be carried out once a day or divided into several doses. The dose and the number of administrations do not limit the scope of the present invention in any aspect.

According to another embodiment of the present invention, there is provided a health functional food for improving or preventing obesity comprising the compound of formula (1) as an active ingredient. Herein, the term " health functional food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be directly eaten through a certain processing step.

Examples of the health functional food to which the compound of the formula (1) of the present invention can be added include various foods, beverages, gums, tea, vitamin complex, and the like. In addition, in the present invention, the food may contain special nutritional foods (eg, crude oil, infant formula, etc.), meat products, fish products, tofu, mackerel, noodles (eg, (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods (For example, fruits and vegetables, beverages, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.). The food, beverage or food additive may be prepared by a conventional production method.

In the present specification, the functional food refers to a food group which is imparted with added value to function and express the function of the food by using physical, biochemical, biotechnological techniques, etc., Means a food which is processed and designed so that the body control function related to restoration and the like is sufficiently expressed to the living body. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In the present specification, beverage means a generic term for drinking or enjoying a taste and is intended to include a functional beverage. The beverage is not particularly limited as long as it contains the compound of formula (1) as an active ingredient as an essential ingredient at the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient can do. Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and Xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention. In addition, the composition of the present invention may be used for natural fruit juice, fruit juice drink, And may further contain flesh for manufacture.

In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not so important, but may be selected in the range of 0 to 20 parts by weight per 100 parts by weight of the compound of formula (1) of the present invention.

In the present specification, functional beverages are used to control the bio-defense rhythm of the beverage group or beverage composition to which the added value is imparted so that the function of the beverage functions for a specific purpose by using physical, biochemical, biotechnological techniques, Means a beverage which has been designed so that the body control function related to restoration and the like is sufficiently expressed in the living body.

The above-mentioned functional beverage is not particularly limited to the other ingredients other than the compound of the above-mentioned formula (1) as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient can do. Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and xylitol , Sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention.

In addition, in the health functional food for the purpose of improving or preventing obesity, the amount of the extract may be 0.01 to 15% by weight of the total food, and the beverage composition may contain 0.02 to 5 g May be contained in a proportion of 0.3 to 1 g.

The composition comprising the compound of formula (1) of the present invention or an alcohol extract of albatrolus pleti mushroom containing the same as an active ingredient inhibits the activity of pancreatic lipase and inhibits digestion and absorption of fat in the body, Can be useful for therapeutic or prophylactic purposes.

FIG. 1 is a graph showing changes in activity of pancreatic lipase using each fraction.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the structure and effects of the present invention will be described in detail with reference to specific embodiments of the present invention. It should be understood, however, that the following examples are provided for a better understanding of the invention and are not to be construed as limiting the scope of the invention.

Hereinafter, the present invention will be described in detail with reference to examples.

1-1 Alcohol Extract Preparation of Albatrolus Pluti Mushroom

Alkyl (50 ml) was mixed with 2 g of dried Albatrellus fletti powder obtained from Gyeongsangbuk-do Forestry Research Institute and extracted with a shaking incubator at 37 ℃ for 24 hours. The supernatant of the shaking incubator was centrifuged at 2500 rpm for 10 minutes, and the alcohol extract of albatrolus pleti mushroom was collected and stored at -20 ° C.

1-2. HPLC analysis Experimental conditions

The reagents used for HPLC analysis were acetonitrile (JT Baker, USA) and tri- acetoacetic acid (Thermo Scientific, USA). AKTA basic (GE, USA) was used as an HPLC system to identify the active substance from the alcohol extract of albatrolus pletifusa mushroom and UV detection (215, 254, 280 nm). The HPLC conditions were as follows: A buffer (0.1% TFA in H2O) and B buffer (0.1% TFA in ACN) were flowed at a retention time of 63 min (B 100%) and 1 ml / min in a gradient manner , A single substance was isolated from albatrolus pleti mushroom alcohol extract using a reversed-phase C18 column (VYDAC? 218TP54 250 X 4.6 mm ID, 5 μm particle) manufactured by GRACE and the purity measurement was also confirmed by HPLC Respectively.

Experimental Example

2-1. Analysis of single substances isolated from alcohol extract of Albatella lupus pleti mushroom

40 mg of albatrolus plutium mushroom alcohol extract was dissolved in 1 ml of DMSO in an Eppendorf tube (AXYGEN Scientific, USA) under the HPLC condition of Example 1-2, and 91 fractions (1 ml / tube) were obtained. The activity of pancreatic lipase was measured using each fraction. The active fractions (E12, F1) exhibiting the highest activity were collected and analyzed for the active substance.

2-2. Measurement and Results of Pancreatic Lipase Activity of Fractions Isolated from Albatrolus pleti mushroom Alcohol Extract

Pancreatic lipase plays a role in recognizing acyl chain-bearing substrates nonspecifically and isolating acyl chains. In order to measure the activity of pancreatic lipase by a single substance isolated from the mushroom alcohol extract of the Examples, porcine pancreatic lipase type 2 (Type II, Sigma chemical) produced by Sigma Chemical Co., Ltd. was used Respectively. Using pancreatic lipase's ability to decompose chromo-genic substrate (p-nirophenylbutyrate), the activity of the enzyme was confirmed through color development. The enzyme inhibition activity was measured by a 405 nm absorbance change using a microplate reader and expressed as a percentage based on the 405 nm absorbance of the fraction-untreated group. The enzymatic activity of 91 fractions isolated from the alcohol extract of albatrolus pleti mushroom was measured and the results showed that the activity of p12 was decreased to 10% or less in the E12 F1 fraction. The results are shown in Fig.

2-3. Mass analysis of single substances isolated from alcohol extracts of albatrolus pleti mushroom

ESI-MS and NMR were used for mass spectrometry of the compounds isolated from the active fractions of FIG. A single material is observed in the ESI-MS to 327.32 [MH], 371.28 [of 655.44 MS / MS], 655.44 [ 2M-H] Molecular weight was found to be 328.24, molecular formula was C ₂₂ H ₃₂ O _2.

2-4. Structural analysis of single substances isolated from alcohol extracts of albatrolus pleti mushroom

The ¹ H-NMR spectrum and the ¹³ C-NMR spectrum were measured at 600 MHz (solvent: CDCl ₃ ) to determine the separated single substance structure, and it was confirmed to be glypholine by mass data.

¹ H-NMR spectrum (600 MHz, _{solvent; CDCl 3): δ (ppm} ) = 6.26 (s, H-1, Aromatic-H), 6.26 (s, H-3, Aromatic-H), 2.24 (s, H-7, Aromatic-H), 3.409-3.421 (d, H-1 ', J = 7.2 Hz), 5.290-5.292 (t, H-2), 1.994-2.008 = 7.2 Hz and 7.8 Hz), 2.069-2.120 (q, H-5 ', J = 7.8 Hz), 5.101-5.114 (q, H- 7.2 Hz and 7.8 Hz), 2.123-2.147 (q, H-9 ', J = 7.8 Hz), 5.101-5.114 (q, H-10), 1.702-1.703 , H-13 '), 1.614 (s, H-14'), 1.844 (s, H-15 '

¹³ C-NMR spectrum (600 MHz, _{solvent; CDCl 3): δ (ppm} ) = 110.36 (C-1), 137.56 (C-2), 109.05 (C-3), 154.82 (C-4), 113.55 ( C-2), 137.56 (C-3), 39.66 (C-4), 139.6 (C- , 26.68 (C-5`), 123.58 (C-6`), 135.62 (C-7`), 39.69 (C-8`), 29.36 (C-11), 26.36 (C-12), 17.69 (C-13), 16.24

Molecular formula: 5-Methyl-2- (3,7,11-trimethyl-2,6,10-dodecatrienyl) -1,3-benzenediol, 9CI. 2-Farnesyl-5-methylresorcinol.

CRC Number: CJT91-X

CAS Registry Number: 6903-07-7

Type of Compound Code (s): DS9900 VS9900 WI3800 ZG4000 ZQ5280

Molecular formula: C ₂₂ H ₃₂ O ₂

Molecular weight: 328.494

Acceleration mass: 328.24023

Composition ratio: C 80.44%; H 9.82%; O 9.74%

Preparation Example: Preparation using glypholin extracted from albatellus pleti mushroom

Production Example 1. Preparation of powder

Glyponin 20 mg, lactose 100 mg, talc 10 mg.

The above ingredients were mixed and filled in an airtight container to prepare powders.

Production Example 2. Preparation of tablets

10 mg of glypholin, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate.

After mixing the above components, tablets were prepared by tableting according to the usual preparation method of tablets.

Production Example 3. Preparation of capsules

10 mg of glypholine, 3 mg of crystalline cellulose, 14.8 mg of lactose, and 0.2 mg of magnesium stearate.

The above components were mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Production Example 4. Production of liquid agent

20 mg of glypholine, 10 g of isomerized sugar, 5 g of mannitol, and purified water.

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Production Example 5. Production of Health Food

1000 mg of glypholine, a proper amount of vitamin mixture, 70 占 퐂 of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.2 mg of vitamin B12, 10 mg of vitamin B12, 10 mg of biotin, Amide 1.7 mg, folic acid 50,, calcium pantothenate 0.5 mg, inorganic mixture suitable amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium phosphate monobasic 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg.

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Claims (20)

delete A pharmaceutical composition for the treatment or prevention of obesity comprising, as an active ingredient, a Grifolin compound represented by the following formula (I):

... (I)
3. The pharmaceutical composition according to claim 2, wherein the compound has a pancreatic lipase activity depressing mechanism. 3. The pharmaceutical composition according to claim 2, wherein the compound has a mechanism of neutral fat absorption inhibition. delete delete delete delete delete delete delete delete delete delete delete The pharmaceutical composition according to any one of claims 2 to 4, wherein the composition is a powder, granule, tablet, suspension, emulsion or syrup. A health functional food composition for prevention and improvement of obesity comprising, as an active ingredient, a Grifolin compound represented by the following formula (I).

... (I) delete A method for treating or preventing obesity comprising the step of administering a Grifolin compound of the formula (I) to an animal other than a human.

... (I)
delete

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