KR101075742B1 - Composition comprising the compounds isolated from the extract of Anemarrhena asphodeloides Bunge for preventing and treating lipid metabolism disorder - Google Patents

Abstract

The present invention relates to a composition containing the compounds isolated from Anemarrhena asphodeloides Bunge as an active ingredient, in detail, since the compounds isolated from the hair have an excellent effect of inhibiting adipocyte differentiation, the composition is effective in preventing lipid metabolism diseases. And it can be usefully used as a therapeutic pharmaceutical composition and health functional food.

Anemarrhena asphodeloides Bunge, Adipocyte Differentiation, Lipid Metabolism Disease

Description

Composition comprising the compounds isolated from the extract of Anemarrhena asphodeloides Bunge for preventing and treating lipid metabolism disorder}

The present invention relates to a composition for the prevention and treatment of lipid metabolism diseases containing compounds isolated from Anemarrhena asphodeloides Bunge having a fat cell differentiation inhibitory effect as an active ingredient.

[1] Park Joong-yeol and 8 others, Changes in Body Weight of Korean Insulin-Independent Diabetic Patients, Diabetes, 17 , pp.51-57, 1993

[Reference 2] Carey VJ. et al., Body fat distribution and risk of non-insulin-dependent diabetes mellitus in women, The nurses' health study, Am J Epidemiol, 145 , pp. 614-619, 1997

Golay A. et al., Obesity and NIDDMP, the retrograde regulation concept, Diabetes Rev , 5 , pp.69-82, 1997

Document 4 Hotamisligil GS. et al., Tumor necrosis factor a, a key component of the obesity-diabetes link, Diabetes , 43 , pp.1271-1278, 1994

[Document 5] Jong Hee Park, Encyclopedia of Herbal Medicine (Ha), Shin Il Sang, pp.746-748, 2002

Preparative isolation and purification of four compounds from the Chinese medicinal herb rhizoma Anemarrhenae by high-speed counter-current chromatography, Journal of chromatography , 1104 , pp.6974, A 2006

Minami E., et al., Stereochemistry of cis- and trans-hinokiresinol and their estrogen-like activity, Chemical & pharmaceutical bulletin, 48 , pp. 389-392, 2000

[8] Takasugi M., et al., Studies on phytoalexins of the Moraceae. 5.Broussonins A and B, new phytoalexins from diseased paper mulberry, Chemistry Letters 3 , pp. 339-340, 1980

[9] Jeong S J., et al., Norlignans with Hyaluronidase Inhibitory Activity From Anemarrhena asphodeloides Planta Med, 65 , pp. 367-368, 1999

Abd El-Hafiz M A., et al., Minor phenolic constituents of Crinum augustum, Journal of Natural Products, 53 , pp. 1349-1352, 1990

11, Camarda L., et al., Dragon's blood from Dracaena draco, structure of novel homoisoflavonoids, Hetero cycles, 20 , pp. 39-43, 1983

12. Namikoshi M., et al., Homoisoflavonoids and related compounds. III. Phenolic constituents of Caesalpinia japonica Sieb. et Zucc., Chemical & Pharmaceutical Bulletin, 35 , pp. 3568-3575, 1987

[13] Matsuda H., et al., Testosterone 5-reductase inhibitory active constituents from Anemarrhenaerhizoma, Biological & Pharmaceutical Bulletin, 24 , pp.586-587, 2001

Herath W., et al., Identification and biological activity of microbial metabolites of xanthohumol, Chemical & Pharmaceutical Bulletin, 51 , pp. 1237-1240, 2003

Barrero A F., et al., Resorcinol Derivatives and Flavonoids of Ononis natrix subspecies ramosissima, Journal of Natural Products, 60 , pp.6568, 1997

16 Ohashi H., et al., 4,4'-Dihydroxychalcone from the heartwood of Chamaecypar isobtusa . Phytochemistry, 27 , pp. 3993-3994, 1988

Obesity is the most common nutritional problem in the industrial community, according to US statistics, one third of Americans and 20% of children are reported as obese, and over 70% of people with type 2 diabetes are obese. There is an increased risk of developing hypertension, hyperlipidemia and some cancers. It is well known that carbohydrate metabolism abnormalities and insulin-independent diabetes mellitus are common in obese people, and that weight gain worsens existing diabetes conditions, but the exact nature of the relationship between obesity and diabetes is still controversial. . Since not all people with severe obesity have diabetes, it is believed that obesity itself is not sufficient to cause insulin-independent diabetes and diabetes is caused by genetic predisposition or environmental factors. However, the incidence of insulin-independent diabetes is much higher in obese people, and since the incidence of insulin-independent diabetes is closely related to the degree and duration of past obesity, obesity is a strong risk factor for insulin-independent diabetes.

On the other hand, not only the degree of obesity but also the distribution of fat is known to be closely related to the disease risk associated with obesity, abdominal fat is a particular problem.

Insulin-independent diabetes patients often have a history of obesity or obesity. In the case of Westerners, 60-90% of patients with insulin-independent diabetes are obese, and in Korea, about 70% of patients with insulin-independent diabetes have a history of obesity (8 patients including Park Joong-yeol and Korean patients with insulin-independent diabetes) Weight change patterns, diabetes mellitus, 17 , pp. 51-57, 1993).

Many prospective studies have also shown that obesity precedes glucose tolerance and insulin-independent diabetes (Carey VJ. Et al., Body fat distribution and risk of non-insulin-dependent diabetes mellitus in women, The nurses' health study, Am J Epidemiol, 145 , pp. 614-619, 1997). In the Framingham study, obesity was found to be more common in obese people, and in most studies, obese people with impaired glucose tolerance were at higher risk for developing insulin-independent diabetes.

The exact mechanism of progression from obesity to diabetes is unknown, but increased insulin resistance is believed to play an important role. Felber et al. Reported that obesity is associated with normal glucose tolerance, impaired glucose tolerance, diabetes with hyperinsulinemia and diabetes with hypoinsulinemia, according to epidemiological observations. It was classified into four groups and progressed from obesity to diabetes (Golay A. et al., Obesity and NIDDMP, the retrograde regulation concept, Diabetes Rev , 5 , pp.69-82, 1997).

In obesity, carbohydrate metabolism is normal in blood glucose response to cervical glucose load, but insulin resistance is higher than normal. Observed from the beginning. Various hypotheses have been suggested as to what mechanisms reduce insulin action in obesity, but it is not yet clear. In other words, the decrease in function of insulin receptors, glucose transporters, enzymes involved in glycogen synthesis and glucose oxidation are observed during obesity, but is understood as a secondary phenomenon due to obesity (Golay A. et al., Obesity and NIDDMP, the retrograde regulation concept, Diabetes Rev , 5 , pp.69-82, 1997).

Given the contextual evidence, it is thought that adipocytes contribute to the development of insulin resistance, that is, factors or metabolic messengers secreted from adipocytes are expected to inhibit the action of insulin in muscle and liver. The first and most widely recognized of these factors is the free fatty acids that are hydrolyzed from triglycerides stored in adipose tissue. In addition, TNF-α and leptin, which are released from adipocytes, have recently been suggested as a cause of insulin resistance in obesity (Hotamisligil GS. Et al., Tumor necrosis factor a, a key component of the obesity-diabetes link, Diabetes , 43 , pp. 1271-1278, 1994).

In obesity, energy metabolism is mainly lipid, and both lipid breakdown and lipid synthesis are increased. Increased lipolysis and thus increased use of free fatty acids are a hallmark of obesity, especially abdominal obesity, and a high correlation between fat volume and lipid oxidation is observed, suggesting that increased fat volume may contribute to increased free fatty acid levels and lipid oxidation. I think. Lipid oxidation is observed in the early stages of obesity, but also on fasting or after glucose load.

In recent years, as the eating habits have been westernized and the culture level has been improved, simple obesity has increased significantly. Obesity has been a cause of involvement or exacerbation of various diseases such as hypertension, arteriosclerosis, fatty liver and diabetes. Therefore, it is better to prevent obesity in advance, and if obesity is to control the body metabolism smoothly through the reduction of body fat and lipid metabolism is very important for the prevention and health of various diseases.

Anemarrhena asphodeloides Bunge is a perennial herb from Liliaceae that uses dried rhizome as a medicine and is distributed in Korea (Hwanghae, Pyeongnam), China, Mongolia, etc. Distributed. Jimo is called 'Jimo' because its root shape is similar to insects. It has a peculiar smell, tastes a little sweet and mucus, and later bitter, effective for diuresis, fever, Jinhae, and Jinhae.

Ingredients are about 6% steroidsaponin, timosaponin A-I, A-II, A-III, A-IV (in the water-insoluble part of methyl alcohol X) and timosaponin ( timosaponin B-I and B-II (in the receptacle), and as a basic saponin, sarsasapogenin, markogenin, and neogitogenin are known. Other xanthones; Mangiferin and isomangiferin are included and vitamintin contains 200µg / g of nicotinic acid. 2002) Known, but none of this document discloses or teaches the lipid metabolism improving effects of compounds isolated from the hair.

Therefore, the present inventors completed the present invention by confirming that the result of measuring the inhibition of adipocyte differentiation using the compounds isolated from the hairs was excellent in improving the adipocyte differentiation, and thus it was excellent in the prevention and treatment of the lipid metabolism disease. .

In order to carry out the above object, the present invention is ( E ) -5,7-dihydroxy-3- (represented by the formula (a), (b) or (c) separated from Anemarrhena asphodeloides Bunge 4'-hydroxybenzylidene) -6-methylchroman-4-one (( E ) -5,7-dihydroxy-3- (4'-hydroxybenzylidene) -6-methylchroman-4-one), 7-hydroxy Hydroxy-3- (4-hydroxybenzyl) coumarin (7-hydroxy-3- (4-hydroxybenzyl) coumarin) or 1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one (1 , 3-bis (4-hydroxyphenyl) prop-2-yn-1-one) novel compound or a pharmaceutically acceptable salt thereof;

                                            (a)

Compounds of formula (a) as defined herein include ( E ) -6-methylurecomalin (( E ) -6-Methyleucomnalin, ( E ) -5,7-dihydroxy-3- (4′-hydroxybenzylidene) -6 -methylchroman-4-one).

                                            (b)

The compound of formula (b) as defined herein is characterized as named Anemarcoumarin A, 7-hydroxy-3- (4-hydroxybenzyl) coumarin.

                                            (c)

Compounds of formula (c) as defined herein are characterized by being named Anemarchalconyn (1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one).

The compound represented by the formula (a), (b) or (c) may be prepared as a pharmaceutically acceptable salt according to a conventional method in the art.

As salts are acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

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

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

Pharmaceutically acceptable salts of the compounds of formula (a), (b) or (c) may be acidic or basic which may be present in compounds of formula (a), (b) or (c) unless otherwise indicated. Salts of groups. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, which are known in the art. It can be prepared through.

The present invention provides a method for the prevention of lipid metabolism disease containing as an active ingredient a novel compound represented by the formula (a), (b) or (c) isolated from Anemarrhena asphodeloides Bunge or a pharmaceutically acceptable salt thereof Provided is a therapeutic pharmaceutical composition.

The invention jimo (Anemarrhena asphodeloides Bunge) 4'- O represented by the following formulas (d) to (n) separated from -methyl California sol (4'- O -Methylnyasol), California sol (Nyasol), Brow sonin A (Broussonin A), 1,3- bis - di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one), 2 ' , 4 ', 4-trihydroxychalcone (2', 4 ', 4-Trihydroxychalcone), 7,4'-dihydroxyhomoisoflavane (7,4'-Dihydroxyhomoisoflavane), 2'- O -methyl 2'- O- Methylisoliquiritigenin, 2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6- Trihydroxy-4-methoxybenzophenone), 4 ', 7- dihydroxy-5-methoxy flavanone (4 ', 7-dihydroxy- 5-methoxyflavanone), 2'- O - methyl Floresta tin (2'- O -Methylphloretin) or 4,4'-dihydroxy chalcone ( It provides a pharmaceutical composition for the prevention and treatment of lipid metabolism disease containing a compound selected from 4,4'-Dihydroxychalcon) as an active ingredient.

In addition, the present invention is a lipid metabolism disease containing a novel compound represented by the formula (a), (b) or (c) or a pharmaceutically acceptable salt thereof isolated from Anemarrhena asphodeloides Bunge as an active ingredient Provide health functional foods for prevention and improvement.

The invention jimo (Anemarrhena asphodeloides Bunge) 4'- O represented by the following formulas (d) to (n) separated from -methyl California sol (4'- O -Methylnyasol), California sol (Nyasol), Brow sonin A (Broussonin A), 1,3- bis - di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one), 2 ' , 4 ', 4-trihydroxychalcone (2', 4 ', 4-Trihydroxychalcone), 7,4'-dihydroxyhomoisoflavane (7,4'-Dihydroxyhomoisoflavane), 2'- O -methyl 2'- O- Methylisoliquiritigenin, 2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6-Trihydroxy-4-methoxybenzophenone), 4 ', 7- dihydroxy-5-methoxy flavanone (4 ', 7-dihydroxy- 5-methoxyflavanone), 2'- O - methyl Floresta tin (2'- O -Methylphloretin) or 4,4'-dihydroxy chalcone ( It provides a health functional food for the prevention and improvement of lipid metabolism disease containing a compound selected from 4,4'-Dihydroxychalcon) as an active ingredient.

4'- O - methyl California sol (4'- O -Methylnyasol)

                                  (d)

Nyasol

                         (e)

Broussonin A

                              (f)

1,3-bis-di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one)

                              (g)

2 ', 4', 4-trihydroxychalcone

                               (h)

7,4'-Dihydroxyhomoisoflavane

                               (i)

2'- O - sound genin Curie Tea methyl (2'- O -Methylisoliquiritigenin)

                               (j)

2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6-Trihydroxy-4-methoxybenzophenone)

                             (k)

4 ', 7-Dihydroxy-5-methoxyflavanone

                            (l)

2'- O - methyl Floresta tin (2'- O -Methylphloretin)

                           (m)

4,4'-Dihydroxychalcone

                           (n)

The lipid metabolism disease is obesity, diabetes mellitus, hypercholesterolemia, hyperlipidemia, hypolipidemia, lipoproteinosis or arteriosclerosis and the like, and preferably includes obesity or diabetes.

Hereinafter, the present invention will be described in detail.

The compound of the present invention is dried hair, preferably the hair is finely divided into about 1 to 20 times, preferably about 1 to 10 times the dry weight of water, C ₁ to C ₄ lower alcohol or a mixed solvent thereof, Preferably with water or methanol, cold extraction, hot water extraction, ultrasonic waves for about 1 hour to 10 days, preferably about 20 hours to 30 hours at room temperature (20 to 100 ℃, preferably 20 to 30 ℃) extraction temperature An extraction method such as extraction and reflux cooling extraction, preferably 1 to 10 times, preferably 2 to 7 times repeated extractions obtained using a reflux cooling extraction method, followed by concentration under reduced pressure to obtain a crude extract; Suspending the crude extract in water and then extracting with ethyl acetate and water, respectively; A third step of diluting the ethyl acetate solvent extract obtained in the second step with n -hexane: ethyl acetate (1-50: 1 (v / v)) by elution with silica gel column chromatography to 25 fractions; A fourth step of dividing the fraction by selecting one of the fractions of the three steps and performing silica gel column chromatography again using chloroform: methanol mixed solvent (1-100: 1 (v / v)); Silica gel column chromatography or Semiprep. Using HPLC it was possible to obtain novel compounds 1 to 3 of the present invention and previously known compounds 4 to 14.

The composition for preventing and treating a lipid metabolism disease of the present invention, the total weight of the composition comprises 0.1 to 50% by weight of the compound.

However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

Compositions comprising a compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The compositions comprising the compounds according to the invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions according to conventional methods. Carriers, excipients and diluents that may be included in the compositions comprising the compounds of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, Alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. . When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The administration may be carried out once a day or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.

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

In addition, the present invention is a lipid metabolic disease containing a novel compound represented by the formula (a), (b) or (c) or a pharmaceutically acceptable salt thereof isolated from Anemarrhena asphodeloides Bunge as an active ingredient Provide health functional foods for prevention and improvement.

The present invention relates to 4'- O -Methylnyasol represented by Formulas (d) to (n) isolated from Anemarrhena asphodeloides Bunge, 4'- O- Methylnyasol, Nyasol, Brosonin A (Broussonin A), 1,3- bis - di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one), 2 ' , 4 ', 4-trihydroxychalcone (2', 4 ', 4-Trihydroxychalcone), 7,4'-dihydroxyhomoisoflavane (7,4'-Dihydroxyhomoisoflavane), 2'- O -methyl 2'- O- Methylisoliquiritigenin, 2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6-Trihydroxy-4-methoxybenzophenone), 4 ', 7- dihydroxy-5-methoxy flavanone (4 ', 7-dihydroxy- 5-methoxyflavanone), 2'- O - methyl Floresta tin (2'- O -Methylphloretin) or 4,4'-dihydroxy chalcone ( It provides a health functional food for the prevention and improvement of lipid metabolism disease containing a compound selected from 4,4'-Dihydroxychalcon) as an active ingredient.

The composition comprising the compound of the present invention can be used in various ways, such as drugs, foods and drinks for the prevention and improvement of lipid metabolism diseases. Examples of the food to which the compound of the present invention may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Since the compound of the present invention has little toxicity and side effects, it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The compound of the present invention may be added to food or beverage for the purpose of preventing and improving lipid metabolism disease. At this time, the amount of the compound in the food or beverage is generally the health food composition of the present invention can be added to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably It can be added at a ratio of 0.3 to 1 g.

The health beverage composition of the present invention, in addition to containing the compound as an essential ingredient in the indicated proportions, has no particular limitation on the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

As described above, the compounds isolated from the hair of the present invention have excellent fat cell differentiation inhibitory effect, and thus can be usefully used as a pharmaceutical composition for preventing and treating lipid metabolism diseases.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Reference Example 1. Compound Structure Determination

The following compounds were evaluated for purity of each component by using analytical TLC, and various spectroscopic IR (Bio-Rad Lavoratoories, USA), Mass (JOEL, Japane) in the state identified as a single active ingredient , ¹ H-NMR (Varian, CA), ¹³ C-NMR, DEPT, HSQC, HMBC, COZY, NOESY, polarimeter (Jasco P-1010), CD (Jasco J-810) and the like Decided.

Example 1 Preparation of Anemarrhena asphodeloides Bunge Crude Extract

20 kg of hairs purchased from Omni Herb.com (www.omniherb.com) were placed in 20 L of 100% methanol and extracted for 24 hours on a water bath using a reflux condenser. This process was repeated three more times to collect the supernatant, and then concentrated under reduced pressure to obtain 4000 g of crude crude extract.

Example 2 Preparation of Geopolar and Nonpolar Solvent Fractions

After 4000 g of the crude crude extract obtained in Example 1 was suspended in 1 L of distilled water, 1 L of hexane was added to dissolve it, and only the components dissolved in the hexane layer were separated and dried in vacuo. This process was repeated 10 times to obtain 80 g of hexane fraction. Left 1 liter of ethyl acetate was added to the aqueous layer, and only the components dissolved in the ethyl acetate layer were separated and dried in vacuo. This procedure was repeated 10 times to obtain 75 g of ethyl acetate fraction. The same procedure as described above gave 180 g of n-butanol fraction and 3665 g of aqueous layer.

Example 3. Compounds Isolated from Hair

3-1. Separation and Purification of (-)-4'-O-Methylazole ((-)-4'-O-methylnyasol)

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 10 g of fraction 8 (F8) of these fractions was subjected to silica gel column chromatography (10: 1 to 10: 1) using a chloroform: methanol mixed solvent (100: 1 to 10: 1). Ø8 × 70 cm, Merk) was separated into five small fractions (F8-1 to F8-5). Three fractions (F8-3) of 10 g of these fractions were subjected to silica gel column chromatography (Ø5 × 60 cm, Merk) again using chloroform: methanol mixed solvent (100: 1 → 10: 1), and 5 Small fractions (F8-3-1 to F8-3-5) were obtained, and then the second fraction (F8-3-2) was extracted from Semiprep. HPLC (YMC-pack ODS-A, Ø2 × 30 cm, Methanol: Water (75:25 v / v)) was carried out and a conventionally known compound represented by the following general formula (4) (Preparative isolation and purification of four Compound 4 (15 mg), which is a compound from the Chinese medicinal herb rhizoma Anemarrhenae by high-speed counter-current chromatography, Journal of chromatography , 1104 , pp.69-74, A 2006), was isolated.

Compound (4): (-)-4'-O- Methylniazole ((-)-4'-O- Methylnyasol)

Colorless resin;

C ₁₈ H ₁₈ O ₂ .

[a] ²⁰ _D = -65.7 ° (c = 0.75, MeOH). CD (MeOH, c = 0.15 × 10 ⁻² M): [θ] ₂₂₈ +12.5, [θ] ₂₅₂ -52.9;

EI-MS: m / z (%) = 266;

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 7.24 (2H, d, J = 8.4 Hz, H-2 ′, 6 ′), 7.12 (2H, d, J = 8.4 Hz, H-2 ″, 6 ″), 6.87 (2H, d, J = 8.8 Hz, H-3 ′, 5 ′), 6.78 (2H, d, J = 8.8 Hz, H-3 ″, 5 ″), 6.54 (1H, d, J = 11.6 Hz, H-1), 6.05 (1H, m, H-4), 5.70 (1H, t, J = 11.4, 10.6 Hz, H-2), 5.14 5.19 (2H, m, H-5), 4.52 (1H, t, J = 9.6, 6.0 Hz, H-3), 3.80 (3H, s, OCH ₃ );

¹³ C-NMR (100 MHz, CDCl ₃ ) δ: 158.7 (s, C-4 ′), 154.2 (s, C-4 ″), 140.9 (d, C-4), 135.8 (s, C-1 ″ ), 131.9 (d, C-2), 130.0 (s, C-1 ′), 129.9 (d, C-2 ′, 6 ′), 129.1 (d, C-2 ″, 6 ″), 128.8 (d , C-1), 115.6 (d, C-3 ″, 5 ″), 115.2 (t, C-5), 113.9 (d, C-3 ′, 5 ′), 55.4 (q, OCH ₃ ), 47.0 (d, C-3).

3-2. Separation and Purification of (-)-Niazole ((-)-Nyasol) and Broussonin A

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 10 g of fraction 8 (F8) of these fractions was subjected to silica gel column chromatography (10: 1 to 10: 1) using a chloroform: methanol mixed solvent (100: 1 to 10: 1). Ø8 × 70 cm, Merk) was separated into five small fractions (F8-1 to F8-5). 10 g of fraction 3 (F8-3) of these fractions was subjected to silica gel column chromatography (Ø5 × 60 cm, Merk) using a chloroform: methanol mixed solvent (100: 1 → 10: 1) again. After obtaining small fractions (F8-3-1 to F8-3-5), the third fraction (F8-3-3) from this was mixed with hexane: ethyl acetate mixed solvent (50: 1 → 10: 1 v / v). ), Silica gel column chromatography (Ø8 × 70 cm, Merk) was used as a developing solvent, and conventionally known compounds represented by the following general formulas (5) to (6) (Minami E., et al. , Stereochemistry of cis- and trans-hinokiresinol and their estrogen-like activity, Chemical & pharmaceutical bulletin, 48 , pp. 389-392, 2000; Takasugi M., et al., Studies on phytoalexins of the Moraceae. 5.Broussonins A and B, new phytoalexins from diseased paper mulberry, Chemistry Letters 3 , pp. 339-340, 1980), compound 5 (2.0 g) and compound 6 (10 mg) were isolated, respectively.

Compound (5): (-)-Niazole ((-)-Nyasol)

Yellow sticky oil

C ₁₇ H ₁₆ O ₂ . [α] ²⁰ _D = -75.3˚ ( c 0.5, MeOH)

IR _max 3369, 1609, 1511, 1460, 1033, 918 cm ^-1

UV _max (log ε)) 207 nm (4.47), 257 nm (4.09)

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 7.18 (2H, d, J = 8.4 Hz, H-2′and H-6 ′), 7.11 (2H, d, J = 8.8 Hz, H-2 ″ and H-6 ″), 6.80 (2H, d, J = 8.8 Hz, H-3 ′ and H-5 ′), 6.78 (1H, d, J = 8.4 Hz, H-3 ″ and H-5 ″) , 6.53 (1H, d, J = 11.2 Hz, H-1), 6.05 (1H, ddd, J = 16.8, 11.2, 6.4 Hz, H-4), 5.70 (1H, t, J = 11.6, 10 Hz, H -2), 5.13-5.18 (2H, m, H-5), 4.51 (1H, dd, J = 6, 10 Hz, H-3);

¹³ C NMR (100 MHz, CDCl ₃ ) δ: 128.8 (C-1), 131.9 (C-2), 47.0 (C-3), 140.9 (C-4), 115.2 (C-5), 130.1 (C -1 ′), 130.2 (C-2 ′ and 6 ′), 115.3 (C-3 ′ and 5 ′), 154.7 (C-4 ′), 135.8 (C-1 ″), 129.1 (C-2 ″ and 6 ″), 115.6 (C-3 ″ and 5 ″), 154.2 (C-4 ″)

LREIMS m / z (% rel. Int.) 252 ([M] ⁺ , 100), 237 (25), 158 (54), 145 (50), 131 22, 107 (58).

Compound (6): Broussonin A

Amorphous solid.

IR _max 3400, 1615, 1590, 1515 cm ^-1

UV _max (log ε) 287.0 nm (3.63), 280.0 nm (3.69), and 225 nm (4.23);

¹ H-NMR (400 MHz, CD ₃ OD) δ: 6.96 (2H, d, J = 8.8 Hz, H-2 ″ and 6 ″), 6.86 (1H, d, J = 8.0 Hz, H-6 ′) , 6.70 (2H, d, J = 8.4 Hz, H-3 "and 5"), 6.38 (1H, d, J = 2.0 Hz, H-3 ′), 6.31 (1H, dd, J = 7.6, 2.0 Hz , H-5 ′), 3.72 (3H, OCH ₃ ), 2.50 (2H, t, J = 7.6 Hz, H-1), 2.49 (2H, t, J = 7.2 Hz, H-3), 1.79 (2H , m, H-2).

¹³ C NMR (100 MHz, CD ₃ OD) δ: 159.8 (s, C-4 ′), 157.7 (s, C-2 ″), 156.2 (s, C-4 ″), 135.0 (s, C-1 ″), 131.2 (d, C-6 ′), 130.4 (d, C-2 ″ and 6 ″), 122.9 (s, C-1 ′), 116.1 (d, C-3 ″ and 5 ″), 107.6 (d, C-5 ″), 99.8 (d, C-3 ′), 55.7 (q, OCH ₃ ), 35.9 (C-3), 33.6 (C-2), 30.3 (C-1); LREIMS m / z (% rel. Int.) 258 ([M] ⁺ , 100), 151 (22), 137 (100), 134 (21), 107 (35), 91 (4), 77 (10) .

3-3. ( E ) -6-Methylleucomaline (( E ) -5,7-dihydroxy-3- (4′-hydroxybenzylidene) -6-methylchroman-4-one), 1,3-bis-di- p -Hydroxyphenyl-4-penten-l-one (1,3-bis-di- p -hydroxyphenyl-4-penten-1-one), 2 ', 4', 4-trihydroxychalcone (2 ', 4', 4-Trihydroxychalcone) and 7,4'-dihydroxyhomoisoflavan (7 Purification of 4, -Dihydroxyhomoisoflavane

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 3 g of fraction 11 (F11) of these fractions was purified by silica gel column chromatography using a chloroform: methanol mixed solvent (50: 1 to 5: 1). Ø8 × 70 cm, Merk) was separated into 16 small fractions (F11-1 to F11-16). Fraction 16 of these fractions (F11-16) was added to Semiprep. HPLC (YMC-pack ODS-A, Ø2 × 30 cm, methanol: water (75:25 v / v)) gave compound 1 (5 mg), which is one novel compound represented by the following general formula (1). ) And previously known compounds represented by the following general formulas (7) to (9) (Jeong S J., et al., Norlignans with Hyaluronidase Inhibitory Activity From Anemarrhena asphodeloides Planta Med, 65 , pp. 367-368, 1999 ; Abd El-Hafiz M A., et al., Minor phenolic constituents of Crinum augustum, Journal of Natural Products, 53 , pp.1349-1352, 1990; Camarda L., et al., Dragon's blood from Dracaena draco, structure Compounds 7 (8 mg), 8 (10 mg) and Compound 9 (2 mg), which are of novel homoisoflavonoids, Hetero cycles, 20 , pp. 39-43, 1983), were isolated, respectively.

Compound (1): ( E ) -5,7-dihydroxy-3- (4′-hydroxybenzylidene) -6-methylchroman-4-one (( E ) -5,7-dihydroxy-3- (4′-hydroxybenzylidene) -6-methylchroman-4-one, or less E ) -6-Methyleucomnalin

Yellow powder.

IR ν _max (KBr) 3368, 2913, 1730, 1595, 1467 cm ⁻¹ ;

UV _max (log) (MeOH) 288 (3.8) nm;

¹ H-NMR (400 MHz, CD ₃ OD) δ: 5.27 (2H, d, J = 1.6 Hz, H-2), 5.88 (1H, s, H-8), 6.87 (2H, dd, J = 2.0, 8.4 Hz, H-3 ', 5'), 7.23 (2H, dd, J = 2.0, 8.4 Hz, H-2 ', 6'), 7.72 (1H, s, H-7 '), 1.95 (3H, s, 6-CH ₃ );

¹³ C-NMR (100 MHz, CD ₃ OD) δ 66.8 (C-2), 128.9 (C-3), 186.6 (C-4), 103.4 (C-4a), 163.5 (C-5), 105.7 ( C-6), 166.3 (C-7), 95.1 (C-8), 161.6 (C-8a), 127.2 (C-1 '), 133.6 (C-2', 6 '), 116.9 (C-3 ′, 5 ′), 160.7 (C-4 ′), 137.9 (C-7 ′), 7.1 (6-CH ₃ );

HRFABMS m / z 299.0922 [M + H] ⁺ (calcd for C ₁₇ H ₁₄ O ₅ , 299.0919).

Compound (7): 1,3-bis-di- p -Hydroxyphenyl-4-penten-l-one (1,3-bis-di- p -hydroxyphenyl-4-penten-1-one)

Amorphous solid.

C ₁₇ H ₁₆ O ₃ .

IR _max 3370, 1653, 1511, 1460 cm ⁻¹ ;

UV _max (log ε) 265 nm (4.09);

¹ H-NMR (400 MHz, CD ₃ OD) δ: 7.85 (2H, d, J = 8.4 Hz, H-2`` and H-6 ''), 7.11 (2H, d, J = 8.8 Hz, H -2 'and H-6'), 6.89 (2H, d, J = 8.4 Hz, H-3 '' and H-5 ''), 6.74 (2H, d, J = 8.8 Hz, H-3 'and H-5 '), 6.03 (1H, m, H-4), 4.95 (2H, m, H-5), 4.00 (1H, q, J = 7.2 Hz, H-3), 3.32 (2H, dd, J = 3.2, 7.2 Hz, H-2);

¹³ C-NMR (100 MHz, CD ₃ OD) δ: 196.0 (C-1), 44.2 (C-2), 45.1 (C-3), 143.1 (C-4), 113.9 (C-5), 135.2 (C-1 '), 139.7 (C-2' and 6 '), 116.0 (C-3' and 5 '), 156.7 (C-4'), 130.6 (C-1 "), 131.4 (C-2 "and 6"), 116.0 (C-3 "and 5"), 162.6 (C-4 ");

EIMS m / z : 268 [M] ⁺ .

Compound (8): 2 ', 4', 4-trihydroxychalcone (2 ', 4', 4-Trihydroxychalcone)

Yellow solid.

C ₁₅ H ₁₂ O ₄ .

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 6.28 (1H, d, J = 2.4 Hz, H-3 ′), 6.41 (1H, dd, J = 2.8, 8.8 Hz, H-5 ′), 6.83 (2H, d, J = 8.4 Hz, H-2, 6), 7.59 (1H, d, J = 15.9 Hz, H-8), 7.62 (2H, d, J = 8.4 Hz, H-3, 5) , 7.77 (1H, d, J = 15.9 Hz, H-7), 7.96 (1H, d, J = 15.9 Hz, H-6 ′);

¹³ C NMR (100 MHz, CDCl ₃ ) δ: 127.9 (C-1), 131.8 (C-2, 6), 116.9 (C-3, 5), 161.6 (C-4), 145.7 (C-7) , 118.4 (C-8), 193.5 (C-9), 114.7 (C-1 '), 167.5 (C-2'), 103.8 (C-3 '), 166.4 (C-4'), 109.2 (C -5 '), 133.4 (C-6').

EIMS m / z : 256 [M] ⁺ .

Compound (9): 7,4'-dihydroxyhomoisoflavan

(7,4'-Dihydroxyhomoisoflavane)

Yellow solid.

C ₁₆ H ₁₄ O ₄ .

UV (MeOH) max (log) 288 (4.03) nm;

IR (KBr) 3350 (br), 1670, 1610, 1520, 1022, 930 cm ⁻¹ ;

EI-MS: m / z (%) = 270 [M] ⁺ .

¹ H-NMR (400 MHz, CD ₃ OD) δ 2.6, 3.0 (2H, m, H-7´), 2.7 (1H, m, H-3), 4.0 (2H, m, H-2), 6.2 ( 1H, d, J = 2.0 Hz, H-8), 6.4 (1H, dd, J = 2.0, 8.8 Hz, H-6), 6.70 (2H, d, J = 8.4 Hz, H-3 ′, 5 ′ ), 7.0 (2H, d, J = 8.4 Hz, H-2 ', 6'), 7.7 (1H, dd, J = 8.8 Hz, H-5);

¹³ C-NMR (100 MHz, CD ₃ OD) δ 70.0 (C-2), 49.0 (C-3), 195.2 (C-4), 130.4 (C-5), 111.2 (C-6), 166.5 (C -7), 103.3 (C-8), 114.6 (C-4a), 165.0 (C-8a), 130.5 (C-1 '), 131.4 (C-2' / C-6 '), 116.2 (C- 3 '/ C-5'), 157.9 (C-4 ').

3-4. 2'- O Methylisosoriquirigenin (2'- O -Methylisoliquiritigenin) 8 mg and 2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6-Trihydroxy-4-methoxybenzophenone)

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 3 g of fraction 11 (F11) of these fractions was purified by silica gel column chromatography using a chloroform: methanol mixed solvent (50: 1 to 5: 1). Ø8 × 70 cm, Merk) was separated into 16 small fractions (F11-1 to F11-16). Fraction 14 (F11-14) of these fractions was recrystallized by using a methanol: chloroform mixed solvent (1: 1). As a result, conventionally known compounds represented by the following general formulas (10) to (11) (Namikoshi M., et al., Homoisoflavonoids and related compounds.III. Phenolic constituents of Caesalpinia japonica Sieb. Et Zucc., Chemical & Pharmaceutical Bulletin, 35 , pp. 3568-3575, 1987; Matsuda H., et al., Compounds 10 (8 mg) and 11 (1.5 g), Testosterone 5-reductase inhibitory active constituents from Anemarrhenaerhizoma, Biological & Pharmaceutical Bulletin, 24 , pp.586-587, 2001, were isolated.

Compound (10): 2'- O Methylisosoriquirigenin (2'- O -Methylisoliquiritigenin)

Yellow solid.

C ₁₆ H ₁₄ O ₄ .

UV (MeOH) max (log) 286 (3.88) nm;

IR (KBr) 3360 (br), 1675, 1620, 1510, 930 cm ⁻¹ ;

EI-MS: m / z (%) = 270 [M] ⁺ .

¹ H-NMR (400 MHz, CD ₃ OD) δ 6.44 (1H, dd, J = 8.4, 2.0 Hz, H-5 ′), 6.51 (1H, d, J = 2.0 Hz, H-3 ′), 6.81 ( 2H, d, J = 8.8 Hz, H-3, 5), 7.38 (1H, d, J = 15.6 Hz, H-8), 7.50 (2H, d, J = 8.4 Hz, H-3), 7.53 ( 1H, d, J = 15.6 Hz, H-7), 7.56 (1H, d, J = 2.0 Hz, H-3 ');

¹³ C-NMR (100 MHz, CD ₃ OD) δ 128.0 (C-1), 131.5 (C-2, 6), 117.0 (C-3, 5), 161.3 (C-4), 144.2 (C-7) , 125.2 (C-8), 193.0 (C-9), 121.9 (C-1 ′), 162.6 (C-2 ′), 100.0 (C-3 ′), 164.6 (C-4 ′), 109.0 (C -5 '), 133.8 (C-6').

Compound (11): 2,4 ', 6-trihydroxy-4-methoxybenzophenone (2,4', 6-Trihydroxy-4-methoxybenzophenone)

Yellow solid.

C ₁₄ H ₁₂ O ₅ .

IR _max (KBr) 3177, 1661, 1249, 831 cm ⁻¹ ;

UV _max (log ε) (MeOH) 298 (4.3) nm;

¹ H-NMR (400 MHz, CD ₃ OD) δ: 5.97 (2H, s, H-3, 5), 6.76 (2H, d, J = 8.9 Hz, H-3 ′, 5 ′), 7.61 (2H , d, J = 8.9 Hz, H-2 ', 6'), 3.77 (3H, s, OCH ₃ );

¹³ C-NMR (100 MHz, CD ₃ OD) δ: 108.1 (C-1), 162.5 (C-2, 6), 94.5 (C-3, 5), 166.1 (C-4), 197.8 (C = O), 133.1 (C-1 ′), 115.2 (C-2 ′, 6 ′), 132.4 (C-3 ′, 5 ′), 162.0 (C-4 ′), 55.9 (OCH ₃ );

EIMS m / z 260 [M] ⁺ .

3-5. Anemarcoumarin A, 4 ', 7-dihydroxy-5-methoxyflavanone and 4'- O Methyl fluoretine (2'- O -Methylphloretin) Purification

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 5 g of fraction 22 (F22) of these fractions was purified by silica gel column chromatography using a chloroform: methanol mixed solvent (50: 1 to 5: 1). Ø10 × 70 cm, from Merk), and the fifth fraction (F22.5) from the Semiprep. HPLC (YMC-pack ODS-A, Ø2 × 30 cm, methanol: water (60:40 v / v)) showed Compound 2 (2), which is one novel compound represented by the following general formula (2). Mg) and previously known compounds represented by the following general formulas (12) to (13) (Herath W., et al., Identification and biological activity of microbial metabolites of xanthohumol, Chemical & Pharmaceutical Bulletin, 51 , pp. 1237-1240, 2003; Barrero A F., et al., Resorcinol Derivatives and Flavonoids of Ononis natrix subspecies ramosissima, Journal of Natural Products, 60 , pp.6568, 1997), Compound 12 (5 mg) and Compound 13 ( 10 mg) were separated respectively.

Compound (2): 7-hydroxy-3- (4-hydroxybenzyl) coumarin (7-hydroxy-3- (4-hydroxybenzyl) coumarin, hereinafter referred to as Anemarcoumarin A)

Yellow powder.

IR ν _max (KBr) 3 298, 2918, 1690, 1610, 1454 cm ⁻¹ ;

UV _max (log) (MeOH) 286 (4.2) nm;

¹ H-NMR (400 MHz, CD ₃ OD) δ: 3.6 (2H, s, H-7 ′), 6.68 (1H, d, J = 2.0 Hz, H-8), 6.73 (2H, d, J = 8.8 Hz, H-3 ′, 5 ′), 6.75 (1H, dd, J = 2.0, 8.4 Hz, H-6), 7.10 (2H, d, J = 8.8 Hz, H-2 ′, 6 ′), 7.32 (1H, d, J = 8.4 Hz, H-5), 7.45 (1H, s, H-3);

¹³ C-NMR (100 MHz, CD ₃ OD) δ 164.3 (C-2), 126.2 (C-3), 141.7 (C-4), 113.8 (C-4a), 130.1 (C-5), 114.5 ( C-6), 162.3 (C-7), 103.1 (C-8), 156.2 (C-8a), 130.6 (C-1 '), 131.3 (C-2', 6 '), 116.5 (C-3 ', 5'), 157.3 (C-4 '), 36.5 (C-7');

HREIMS m / z 268.0738 [M] ⁺ (calcd for C ₁₆ H ₁₂ O ₄ , 268.0735).

Compound (12): 4 ', 7-dihydroxy-5-methoxyflavanone

Yellow powder.

C ₁₆ H ₁₄ O ₅ .

EI-MS: m / z (%) = 286;

IR ν _max (KBr) 3300, 1670, 1620, 1510 cm ⁻¹ ;

UV _max (log) (MeOH) 260 (4.0) nm;

¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.10 (2H, m, H-3), 5.31 (1H, dd, J = 2.8, 12.8 Hz, H-2), 5.94 (1H, d, J = 2.4 Hz, H-8), 6.04 (1H, d, J = 2.4 Hz, H-6), 6.76 (2H, d, J = 8.4 Hz, H-3 ', 5'), 7.27 (2H, d, J = 8.4 Hz, H-2 '/ H-6'), 3.73 (3H, OMe-5);

¹³ C-NMR (100 MHz, CDCl ₃ ) δ 78.0 (C-2), 44.7 (C-3), 187.7 (C-4), 162.1 (C-5), 93.1 (C-6), 164.2 (C- 7), 95.5 (C-8), 104.0 (C-4a), 164.2 (C-8a), 129.3 (C-1 '), 128.0 (C-2' / C-6 '), 115.0 (C-3 '/ C-5'), 157.5 (C-4 '), 55.7 (OMe).

Compound (13): 2'- O Methyl fluoretine (2'- O -Methylphloretin)

Yellow solid.

C ₁₆ H ₁₆ O ₅ .

UV (MeOH) max (log) 288 (3.98) nm;

IR (KBr) 3400 (br), 1670, 1620, 1510, 930 cm ⁻¹ ;

EI-MS: m / z (%) = 288 [M] ⁺ .

¹ H-NMR (400 MHz, CD ₃ OD) δ 5.88 (1H, dd, J = 8.4, 2.0 Hz, H-5 ′), 5.95 (1H, d, J = 2.0 Hz, H-3 ′), 6.68 ( 2H, d, J = 8.8 Hz, H-3 / 5), 7.02 (2H, d, J = 8.8 Hz, H-2 / 6), 2.83, 3.15 (2H, m, H-7), 3.2 (2H , m, H-8);

¹³ C-NMR (100 MHz, CD ₃ OD) δ 133.9 (C-1), 130.4 (C-2, 6), 116.3 (C-3, 5), 156.6 (C-4), 31.5 (C-7) , 47.5 (C-8), 206.1 (C-9), 106.1 (C-1 '), 165.0 (C-2'), 92.2 (C-3 '), 168.4 (C-4'), 97.1 (C -5 '), 166.6 (C-6').

3-6. Separation and Purification of 1Anemarchalconyn and 4,4'-Dihydroxychalcon

75 g of the ethyl acetate fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (Ø15 × 70 cm, Merk) using a hexane: ethyl acetate mixed solvent (50: 1 → 1: 1 v / v). G) to separate 25 fractions (F1 to F25), and 5 g of fraction 22 (F22) of these fractions was purified by silica gel column chromatography using a chloroform: methanol mixed solvent (50: 1 to 5: 1). Ø10 × 70 cm, Merk) and the sixth fraction (F22.6) from the Semiprep. HPLC (YMC-pack ODS-A, Ø2 × 30 cm, methanol: water (40:60 v / v)) showed that Compound 3 (2), which was one novel compound represented by the following general formula (3): Mg) and a previously known compound represented by the following general formula (14) (Ohashi H., et al., 4,4'-Dihydroxychalcone from the heartwood of Chamaecypar isobtusa.Phytochemistry , 27 , pp. 3993-3994, 1988 ), Compound 14 (5 mg) was isolated, respectively.

Compound (3): 1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one (1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one, hereinafter referred to as Anemarchalconyn Naming)

Yellow powder.

UV (MeOH) λ _max (log e ) 286 (4.2) nm;

IR ν _max (KBr) 3350, 2193, 1620, 1159 cm ⁻¹ ;

¹ H-NMR (400 MHz, CD ₃ OD) δ: 6.85 (2H, d, J = 8.8 Hz, H-3 ′ ′, 5 ′ ′), 6.90 (2H, d, J = 8.8 Hz, H-3 ′, 5 ′), 7.55 (2H, d, J = 8.8 Hz, H-2 ′ ′, 6 ′ ′), 8.07 (2H, d, J = 8.8 Hz, H-2 ′, 6 ′);

¹³ C-NMR (100 MHz, CD ₃ OD) δ: 178.5 (C-1), 87.2 (C-2), 95.7 (C-3), 130.3 (C-1 ′), 133.2 (C-2 ′, 6 ′), 116.6 (C-3 ′, 5 ′), 165.2 (C-4 ′), 111.5 (C-1 ′ ′), 136.3 (C-2 ′ ′, 6 ′ ′), 117.1 (C-3 ′) ′, 5 ′ ′), 161.8 (C-4 ′ ′);

HREIMS m / z 238.0630 [M] ⁺ (calcd for C ₁₅ H ₁₀ O ₃ , 238.0630).

Compound (14): 4,4'-Dihydroxychalcon

Yellow solid.

C ₁₅ H ₁₂ O ₃ .

UV (MeOH) max (log) 234, 350 (3.98) nm;

IR (KBr) 3500 (br), 1750, 1655 cm ⁻¹ ;

EI-MS: m / z (%) = 240 [M] ⁺ .

¹ H-NMR (400 MHz, CD ₃ OD) δ 6.83 (2H, d, J = 8.4 Hz, H-3 / 5), 6.88 (2H, d, J = 8.4 Hz, H-3 ′ / 5 ′), 7.55 (2H, d, J = 8.4 Hz, H-2 / 6), 7.99 (2H, d, J = 8.4 Hz, H-2 '/ 6'), 7.61 (1H, d, J = 16.0 Hz, H -7), 7.68 (1 H, d, J = 16.0 Hz, H-8);

¹³ C-NMR (100 MHz, CD ₃ OD) δ 132.4 (C-1), 129.8 (C-2, 6), 121.9 (C-3, 5), 156.6 (C-4), 125.3 (C-7) , 145.0 (C-8), 190.5 (C-9), 134.2 (C-1 '), 130.1 (C-2' / 6 '), 120.0 (C-3' / 5 '), 168.4 (C-4 ′).

Reference Example 2. 0.3% Oil-red O Staining Solution Preparation

Dissolve the powder in 100% isopropanol to 0.3% Oil-red O, and then continue stirring for about 16 hours for good dissolution. 0.3% Oil-red O: distilled water = 6: 4 (v / v) to the distilled water and mix again, and then filtered once with 3M paper, filtered using a 0.2㎛ syringe filter (syringe filter).

Experimental Example 1. Adipocyte differentiation inhibitory effect

Pre-adipocytes, 3T3-L1 cells (American Tissue Culture.Collection, ATCC) containing 10% bovine calf serum, DMEM (Dulbecco's Modified Eagle Medium, Gibco-BRL, Grand Island, NY, USA) and then seeded by suspending 4 x 10 ⁴ cells per well in 0.5 ml in a 24-well plate. Incubate for 48 hours in the same medium, then reach 100% with 5 μg / ml insulin, 10 μM rosiglitazone, 1 μM dexamethasone, 10% Fetal Bovine Serum Change to DMEM medium and induce differentiation into mature adipocytes. At this time, after dissolving the compounds obtained in Examples 3-1 to 3-6 in DMSO and treated together so that the same concentration of DMSO is used in each concentration treatment group, as a control to control the differentiation into mature fat cells and diabetes Resveratrol, Sigma-ALdrich, 3,4 ′, 5-Trihydroxy-trans-stilbene, 5-[(1E) -2- (4-Hydroxyphenyl) ethenyl] -1,3-benzenediol Is dissolved in DMSO and treated at the same concentration. 48 hours after the induction of differentiation, the drug was again treated with 10% FBS / DMEM medium containing 5 μg / ml insulin (insulin) again, and after 48 hours, it was again changed to 10% FBS / DMEM medium. When 7 days have started to differentiate into mature adipocytes. Identify the fat produced by oil-red O staining. Remove the cell culture solution and wash the cells once with PBS. Be careful not to add too much PBS. 10% formalin was added and the cells were fixed at room temperature for 1 hour, 10% formalin was removed and washed twice with distilled water, followed by adding the Oil-red O staining reagent prepared in Reference Example 2 at room temperature. Stain for 1 hour. After dyeing, the dyeing reagent is removed, washed with distilled water, opened by drying the lid, and then scanned for image. 100% isopropanol is added to the stained cells, and the dyeing reagent is dissolved for 10 minutes. 500 nm Absorbance was measured at the wavelength and observed by comparing the degree of oil-red O staining between the compound and the control group. The IC ₅₀ value was used as a computer (BioDataFit 1.02) to quantify the effect of each compound on adipocyte differentiation.

Experiments, drugs which inhibit the differentiation of adipocytes (adipocyte) as shown in the following Table 1 are 4'- O - methyl California sol (4'- O -Methylnyasol), 2'- O - methyl sound Curie tea genin (2'- O- Methylisoliquiritigenin), brosonin A (Broussonin A), 2 ', 4', 4-trihydroxychalcone (2 ', 4', 4-Trihydroxychalcone), especially analkalcones Phosphorus (Anemarchalconyn) showed a strong adipocyte differentiation inhibitory effect at a lower concentration than Resveratrol used as a control, it was confirmed that there is an excellent effect of lipid metabolism disease.

Adipocyte differentiation inhibition rate of isolated compounds (IC ₅₀ ) compound IC ₅₀ Compound 1 ( E ) -6-Methyleucomnalin 336.1 Compound 2 Anemarcoumarin a nd Compound 3 Anemarchalconyn 5.3 Compound 4 4'- O -Methylnyasol 45.9 Compound 5 Nyasol 107.8 Compound 6 Broussonin a 74.5 Compound 7 1,3-bis-di- p -hydroxyphenyl-4-penten-1-one 326.2 Compound 8 2 ', 4', 4-Trihydroxychalcone 96.4 Compound 9 7,4'-Dihydroxyhomoisoflavane > 500 Compound 10 2'- O -Methylisoliquiritigenin 41.8 Compound 11 2,4 ', 6-Trihydroxy-4-methoxybenzophenone nd ^b Compound 12 4 ', 7-Dihydroxy-5-methoxyflavanone 259.7 Compound 13 2'- O -Methylphloretin > 500 Compound 14 4,4'-Dihydroxychalcon > 500 Positive Standard
(Control group) Resveratrol ^a 31.4 ^a positive control
^{b not} detected

Examples of the pharmaceutical compositions containing the compounds of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Formulation Example 1 Preparation of Powder

( E ) -6-Methyleucominin (( E ) -6-Methyleucomnalin) 20 mg

Lactose 100 mg

Talc 10 mg

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

Formulation Example 2 Preparation of Tablet

Anemarcoumarin A 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Anemarchalconyn 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

 Formulation Example 4 Preparation of Injection

Broussonin A 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

4'- O - methyl California sol (4'- O -Methylnyasol) 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, the lemon flavor is added, the above components are mixed, the purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

 Formulation Example 6 Preparation of Healthy Drink

2 ', 4', 4-trihydroxychalcone 100 mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

Vitamin B ₁ 0.25 g

Vitamin B ₂ 0.3 g

Water quantification

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

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

Claims (7)

( E ) -5,7-dihydroxy-3- (4′-hydroxybenzylidene) -6-methylchroman-4-one (( E ) -5,7 isolated from Anemarrhena asphodeloides Bunge) -dihydroxy-3- (4′-hydroxybenzylidene) -6-methylchroman-4-one), 7-hydroxy-3- (4-hydroxybenzyl) coumarin (7-hydroxy-3- (4-hydroxybenzyl) coumarin) Or 1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one (1,3-bis (4-hydroxyphenyl) prop-2-yn-1-one) novel compound or a pharmaceutical thereof Acceptable salts. A pharmaceutical composition for the prevention and treatment of lipid metabolism diseases comprising the novel compound of claim 1 isolated from Anemarrhena asphodeloides Bunge or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition of claim 2, wherein the lipid metabolism disease is obesity, diabetes mellitus, hypercholesterolemia, hyperlipidemia, hypolipidemia, lipoproteinosis or arteriosclerosis. Jimo the grooving sonin A (Broussonin A) separated from the (Anemarrhena asphodeloides Bunge), 1,3- bis - di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one), 2 ', 4', 4-trihydroxychalcone (2 ', 4', 4-Trihydroxychalcone), 7,4'-dihydroxy homoisoflavan (7,4 '-Dihydroxyhomoisoflavane), 2'- O - methyl sound Curie tea genin (2'- O -Methylisoliquiritigenin), 2,4', 6-trihydroxy-4-methoxybenzophenone (2,4 ', 6 Trihydroxy-4-methoxybenzophenone), 4 ', 7- dihydroxy-5-methoxy flavanone (4', 7-dihydroxy- 5-methoxyflavanone), 2'- O - methyl Floresta tin (2'- O -Methylphloretin ) Or 4,4'-Dihydroxychalcon (4,4'-Dihydroxychalcon) A pharmaceutical composition for the prevention and treatment of obesity containing as an active ingredient. Health functional food for the prevention and improvement of lipid metabolism disease containing the novel compound of claim 1 or a pharmaceutically acceptable salt thereof isolated from Anemarrhena asphodeloides Bunge as an active ingredient. Jimo the grooving sonin A (Broussonin A) separated from the (Anemarrhena asphodeloides Bunge), 1,3- bis - di-p-hydroxy phenyl-4-penten-1-one (1,3-bis-di- p -hydroxyphenyl -4-penten-1-one), 2 ', 4', 4-trihydroxychalcone (2 ', 4', 4-Trihydroxychalcone), 7,4'-dihydroxy homoisoflavan (7,4 '-Dihydroxyhomoisoflavane), 2'- O - methyl sound Curie tea genin (2'- O -Methylisoliquiritigenin), 2,4', 6-trihydroxy-4-methoxybenzophenone (2,4 ', 6 Trihydroxy-4-methoxybenzophenone), 4 ', 7- dihydroxy-5-methoxy flavanone (4', 7-dihydroxy- 5-methoxyflavanone), 2'- O - methyl Floresta tin (2'- O -Methylphloretin ) Or 4,4'-Dihydroxychalcon (4,4'-Dihydroxychalcon) A health functional food for the prevention and improvement of obesity diseases containing a compound selected as an active ingredient. The dietary supplement according to claim 5 or 6 in the form of a powder, granule, tablet, capsule or beverage.