KR101651105B1 - ISOLATED SINGLE COMPOUND FROM Mori Cortex Radicis ITS APPLICATION IN TREATING AND PREVENTING OBESITY - Google Patents

Abstract

The present invention relates to new compounds derived from bovine alfalfa and their uses, and more particularly to novel compounds derived from alcoholic extracts of bovine alfalfa and their use for the treatment and prevention of obesity.

Description

TECHNICAL FIELD OF THE INVENTION [0001] The present invention relates to a composition for treating or preventing obesity of MAC 9.23,

The present invention relates to new compounds derived from bovine alfalfa and their uses, and more particularly to novel compounds derived from alcoholic extracts of bovine alfalfa and their use 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.

Korean Patent Laid-Open Publication No. 10-2013-0065086 discloses the substitution of Xenical ^TM (orlistat) by using an ethanol extract of a flower shield mushroom.

On the other hand, it is disclosed in Korean Patent Application No. 10-2011-0040558 as a composition for treating pancreatic cancer in the form of an extract composition, and is disclosed in Korean Patent Application No. 10-2013-0112012 as a composition for preventing or treating depression.

Manganese berry is a foliage of mulberry plants belonging to the Moraceae family, including mulberry, kuwanon, mulberrofuran, albafuran, morucinon, sanggennon, it contains a large amount of flavonoids such as cholin, stilbene, benzofuran, oxydihydromorusin and the like. In addition, amyrin, betulinic acid, sitosterol ), Umbelliferone, and the like. Along with antioxidant, anticonvulsant, anti-allergic, anti-diabetic and anti-inflammatory effects, it has recently been shown that there is a liver protecting effect and hair growth effect on alopecia, whitening, antioxidation, It is known.

A problem to be solved by the present invention is to provide a novel compound which has not been disclosed.

Another object to be solved by the present invention is to provide a new compound derived from the bark of the bark.

Another object to be solved by the present invention is to provide a medicinal use of a new compound derived from bovine alfalfa.

Another object to be solved by the present invention is to provide a compound for the treatment or prevention of obesity derived from bovine alfalfa.

In order to achieve the above object, the present invention provides a compound of the formula (1) or a salt thereof.

(I)

(I)

In addition, the present invention provides an extract of Morus alba L. comprising the compound of formula (I) and / or a salt thereof.

The present invention also provides a composition for the treatment or prevention of obesity, which comprises the extract of Morus alba Fc. Comprising the compound of formula (1), the salt thereof or the compound of formula (I) and / or a salt thereof as an active ingredient.

The present invention also provides a medicament for the treatment or prevention of obesity, which comprises the extract of Morus alba L., which comprises the compound of the above formula (1), its salt or the compound of formula (I) and / or its salt 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 a bark of phornicus comprising the compound of the above formula (1), a salt thereof, or a compound of the formula (I) and / do.

The present invention also provides a health functional food for improving or preventing obesity, which comprises an extract of bovine bark extract containing a compound of the above formula (1), a salt thereof, or a compound of formula (I) and / or a salt thereof.

The present invention also relates to a method for the treatment and / or prophylaxis of diseases caused by the activity of pancreatic lipase (triglyceride lipase (TGL)), which comprises extracting the extract of Morbuschii comprising the compound of the above formula (1), its salt or the compound of formula (I) and / And can be used for treating or preventing obesity.

The present invention also provides a method for extracting a composition for preventing or treating obesity comprising the compound of the above formula (1) or the compound of the above formula (1) in an alfalfa by using an alcohol having 1 to 10 carbon atoms. The alcohol is preferably a lower alcohol having 1 to 5 carbon atoms, and most preferably methanol.

In the present invention, the alcoholic extract of the phytophthora bark containing the compound of the formula (1) may be prepared by a conventional method for producing a plant extract. Preferably, after removing the residues after drying and crushing the above-mentioned bovine perilla, 0.1 to 20 g of the pulverized product per 100 ml of the alcohol having 1 to 10 carbon atoms is added, more preferably 1 to 5 g of pulverized product per 100 ml of the extraction solvent And then extracted. When the content of the crushed cornflakes is too small as compared with the extraction solvent, the effect of the claryphobia is not sufficient and it is not preferable. If the amount of the crushed material is too large relative to the amount of the extraction solvent, Which is undesirable in view of the above. In this case, the extraction conditions are not limited, but preferably the mixture is mixed with an alcohol having 1 to 10 carbon atoms as an extraction solvent and then heated at a temperature of 20 to 60 DEG C for 12 to 36 hours, more preferably 30 to 40 DEG C RTI ID = 0.0 > 20-24 < / RTI > 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 alcoholic extract of the alfalfa extract, which is extracted by the above method, is filtered through a filter or the like, and the filtrate is centrifuged to remove the precipitate, followed by concentration under reduced pressure or concentration and freeze-drying.

In the present invention, the compound of the above formula (1) can be separated from the alcoholic extract of the above-mentioned Alfalfa by using 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 alcoholic extract of corn alcohols 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. The medicament containing the compound of the above formula (1) as an active ingredient may further contain an appropriate carrier, excipient and diluent 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.

According to the present invention, a novel natural compound such as the above-mentioned formula (1) and a method of separating it from the mantle are disclosed. The composition comprising the compound of the above formula (1) or the alcoholic extract of bovine alfalfa as an active ingredient inhibits the activity of the pancreatic lipase and inhibits digestion and absorption of fat in the body to be useful for the treatment or prevention of obesity .

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process diagram for separating and purifying a compound according to the present invention from an extract of cornucopia. FIG.
Fig. 2 is a reverse phase chromatography of a new single substance MAC 9.23 isolated from bovine alfalfa. The HPLC analysis conditions of MAC 9.23 are as follows.
- HPLC System: Waters Alliance
Column: Agilent Eclipse XDB-C18 (5 [mu] m, 4.6 mm id x 160 mm length)
- mobile phase conditions: methanol: water (60:40 → 100: 0, gradient)
- Analysis time: 60 minutes
- Sample injection amount: 20 μL
- Flow rate: 1 mL / min
- Detection wavelength: UV 265 nm
- Retention time of compound 1: 31.24 min
Figure 3 is the EI-MS spectrum of a new single substance MAC 9.23 isolated from the bark of alfalfa.
Figure 4 is the HR-EI-MS spectrum of a new single substance MAC 9.23 isolated from bovine moss.
5 is the 1 H-NMR data of the new single substance MAC 9.23 isolated from the alfalfa.
FIG. 6 is C-NMR data of a new single substance MAC 9.23 isolated from E.coli.
7 is a structural formula of a new single substance isolated from the bark of alfalfa.
Fig. 8 is a graph showing pancreatic lipase inhibitory activity of the new single substance MAC 9.23 isolated from the bovine bark.
FIG. 9 is a graph showing the measurement of neutral fat absorption inhibition of a new single substance MAC 9.23 isolated from C. bovis.

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.

Example 1

Separation of the active substance from the alfalfa extract

As shown in Figs. 1 and 2, phytomassified morulae ( Morus alba ) was commercially obtained. 15 kg of methanol was added to a finely chopped dried 9 kg of Sanghwabok sample, and the mixture was extracted by cooling with ice three times at room temperature for 3 days. The methanol extract was filtered and concentrated under reduced pressure at 40 ° C to obtain an extract. This was suspended in water, and the suspension was subjected to solvent fractionation successively using hexane, chloroform and ethyl acetate. The pancreatic lipase inhibitory activity was confirmed in chloroform and ethyl acetate fractions in these solvent fractions. Chloroform fraction (214 g) showing strong physiological activity was first subjected to chromatography (CC) using a silica gel column. A total of 19 small fractions were obtained by selecting dichloromethane-methanol as the eluting solvent and gradient elution (100: 0 → 0: 100). Silica gel CC was applied to fraction C3 (12.2 g) . The fraction 17 (C3.17, 0.89 g) of the small fraction thus separated was subjected to gradient elution (RP-18 CC: mobile phase: acetonitrile - water (1: 1)), finally obtaining 95 mg of MAC 9.23.

The physicochemical properties of the compound 1 obtained in the present invention are as follows.

Appearance: Brown colored oil

EI-MS: m / z 446 [M] ^< ⁺ & gt ^;

HR-EI-MS: m / z 446.25 (calculated for C ₂₉ H ₃₄ O ₄ )

Mass spectrometry (MS) revealed molecular weights and elemental structures of the invented compounds through low resolution EI-MS and high resolution HR-EI-MS analysis, and the results are shown in FIG. 3 and FIG. Nuclear magnetic resonance (NMR) analysis was performed at 500 MHz ( ¹ H) and 125 MHz ( ¹³ C) and the chemical shift of each signal was compared with the chemical shift value (0 ppm) of the reference TMS Respectively. The analyzed ¹ H- and ¹³ C-NMR spectra are shown in FIG. 5 and FIG. 6. In addition, the structures determined through the above-described various spectroscopic analyzes are shown in FIG. 7, and NMR spectral data providing crucial information for the structure determination are shown in Table 1. Compound 1 was determined by 2 ', 6', 7'-trihydroxyl-4'-prenyl-6-geranyl-2-arylbenzofuran through various spectroscopic analyzes such as NMR and MS.

Position MAC9.23 隆_H ( J in Hz) δc 2 154.1 3 6.68 (1H, brs) 105.8 3a 122.3 4 6.78, (1H, d, 8.0) 113.0 5 7.26, (1H, d, 8.0) 118.8 6 110.6 7 152.5 7a 154.5 One' 132.4 2' 154.8 3 ' 6.76, (1H, d, 3.0) 108.6 4' 118.0 5 ' 7.41, (1H, d, 2.5) 104.0 6 ' 156.4 8 3.67 (2H, d, 7.0) 23.3 9 5.39 (1H, t, 7.0) 121.2 10 139.1 11 1.83 (3 H, s) 16.4 12 2.07 (2 H, m) 39.8 13 2.05 (2H, m) 26.6 14 5.05 (1H, t, 6.5) 124.0 15 132.2 16 1.56 (3 H, s) 17.8 17 1.64 (3 H, s) 25.9 7 ' 3.51 (2H, d, 6.5) 26.6 8' 5.27 (1H, t, 6.5) 122.8 9 ' 134.7 10 ' 1.76 (3 H, s) 18.1 11 ' 1.74 (3 H, s) 25.8

Example 2. Measurement and Results of Pancreatic Lipase Inhibition Activity of New Single Substance MAC9.23 Isolated from Mushroom

Pancreatic lipase plays a role in recognizing acyl chain-bearing substrates nonspecifically and separating acyl chains. To measure pancreatic lipase activity by the novel single substance isolated from the bovine periwinkle of Example, porcine pancreatic lipase type 2 (Type II, Sigma chemical) manufactured by Sigma Chemical Co. was used as an enzyme. Using the property of pancreatic lipase to degrade chromogenic substrate (p-nirophenylbutyrate), we confirmed the enzyme activity through color reaction. The enzyme inhibitory 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 compound MAC 9.23 untreated group. We measured the enzyme activity of the new single substance MAC9.23 isolated from bovine alfalfa and found that the concentration of active substance (IC50), which inhibits pancreatic lipase activity by 50%, was obtained in MAC9.23 compared to the fraction - untreated group. The results are shown in Fig.

Example 3 Measurement of Neutral Fat Absorption Inhibition of New Single Substance MAC9.23 Isolated from Mushroom

The plasma triglyceride was measured to determine whether the new single substance MAC9.23 isolated from the alfalfa extract showing the inhibitory effect of pancreatic lipase activity was also effective at the individual level. A 0.5 ml oral fat suspension was orally administered to 8-week-old Balb / c mice (male, body weight ~ 25 g) fasted for 16 hours. The fat suspension consisted of 30% olive oil, 0.5% (w / v) carboxymethylcellulose sodium (CMC-Na) and 1% (v / v) Tween 80 (Tween 80) 50 ug (2 mg / kg) of new single substance MAC9.23 was added. After 2 hours, mouse blood was collected and centrifuged at 3000 rpm for 15 minutes at 4 ° C. The amount of triacylglycerol in the serum was measured using a serum triglyceride determination kit (Sigma). As a result, it was confirmed that the new single substance MAC9.23 isolated from the bark of the example lowered the neutral fat absorption. The results are briefly shown in Fig.

Preparation Example: Preparation using MAC9.23 extracted from alfalfa

Production Example 1. Preparation of powder

MAC 9.23 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

MAC 9.23 10 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg.

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 MAC 9.23, 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 MAC 9.23, 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

MAC 9.23 1000 mg Vitamin A acetate 70, Vitamin E 0.13 mg Vitamin B 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 mg Vitamin C 10 mg Biotin 10 mg, 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 (17)

A compound represented by the following formula (1) or a salt thereof.

(I) A method for extracting a compound represented by the following formula (I) or a salt thereof in an alfalfa.

(I) 3. The method according to claim 2, wherein the extract is obtained from alfalfa with an alcohol having 1-10 carbon atoms. 4. The method of claim 3, wherein the alcohol is methanol. A compound for the treatment or prophylaxis of obesity, represented by the following formula (I), or a salt thereof.

(I) 6. The compound according to claim 5, wherein the compound of formula (I) inhibits the absorption of fat. The compound for treating or preventing obesity according to claim 5, wherein the compound of formula (I) inhibits the absorption of fat by lowering the activity of pancreatic lipase. The compound for treating or preventing obesity according to claim 5, wherein the compound of formula (I) is derived from bovine bark. The compound for treating or preventing obesity according to claim 8, wherein the compound of formula (I) is extracted using alcohol. A medicament for treating or preventing obesity comprising a compound represented by the following formula (1) as an active ingredient.

(I) 11. The medicament according to claim 10, wherein the medicament is a powder, a granule, a tablet, a suspension, an emulsion, or a syrup. A health functional food for improving or preventing obesity comprising the compound of formula (I) as an active ingredient.

(I)
13. The health functional food for obesity improvement or prevention according to claim 12, wherein the health functional food is a food, a food additive, or a beverage.
A method of treating or preventing obesity comprising administering a pharmaceutically effective amount of a compound of formula (I) to an animal other than a human.

(I) delete delete delete

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