KR100733764B1 - Composition comprising the cortex extract of albizzia julibrissin or kuraridinol isolated therefrom for preventing or treating hyperlipidemia - Google Patents

Abstract

A composition for preventing or treating hyperlipidemia comprising the extract of Albizzia julibrissin cortex or kuraridinol isolated therefrom is provided to reduce the amounts of the total cholesterol and neutral lipid in blood. The pharmaceutical composition for preventing or treating hyperlipidemia comprises 0.1-50 wt.% of the extract of Albizzia julibrissin cortex or kuraridinol isolated therefrom and represented by the formula(1) as effective ingredients, and pharmaceutically acceptable carriers or excipients, wherein the Albizzia julibrissin cortex extract is prepared by extracting Albizzia julibrissin cortex with a polar solvent selected from water, ethanol, methanol and butanol, or a nonpolar solvent selected from dichloromethane, chloroform and ethylacetate.

Description

Composition comprising the cortex extract of Albizzia julibrissin or kuraridinol isolated therefrom for preventing or treating hyperlipidemia}

The present invention relates to a pharmaceutical composition for the prevention and treatment of hyperlipidemia, which contains haphwanpi extract or kuraridinol isolated therefrom.

Hyperlipidemia refers to a condition in which lipid components in the blood are more than normal (Kim JK, Clinical Geology, pp241-270, 1995). The major components of serum lipids are composed of lipid-soluble substances such as cholesterol, triglycerides, phospholipids, and free fatty acids, and depending on which of these lipids increases, hypercholesterinemia, hypertriglyceridemia, etc. It is called. Increased serum cholesterol and triglycerides are the most common causes of hyperlipidemia. Accumulation of excess fat causes blood circulation disorders and microcirculation failure, resulting in atherosclerosis, ischemic heart disease, cerebral infarction, hypertension, obesity and diabetes. (Schoen FJ et al ., Robbins pathologic basis of disease , 5th ed., Pp 473-484, 1994).

There are many causes of atherosclerosis, but elevated plasma lipids, particularly plasma cholesterol and triglycerides, are known as one of the most important risk factors.

Cholesterol is produced mainly in the liver and is present in the blood in the form of small round particles called lipoproteins, which carry cholesterol throughout the body. There are two types of lipoproteins, low density lipo protein (LDL) and high density lipoprotein (HDL) .Low-density lipoproteins carry cholesterol from other tissues to the liver. In contrast, low-density lipoproteins are mainly produced by the liver and carry cholesterol to other tissues of the human body. Therefore, a lot of low-density lipoproteins accumulate a lot of cholesterol in blood vessels, which promotes arteriosclerosis. .

In fact, cholesterol is an essential ingredient for normal functioning of the human body, a nutrient necessary for making cells, and a component of various hormones such as corticosteroids, male hormones, and female hormones. do. In addition, cholesterol is a nutrient necessary for the human body such as being used as a constituent of biological membranes and as a starting material for hormonal synthesis. However, excessive cholesterol intake is known to accumulate in the blood vessels and cause heart disease, and there is no preventive method except low-cholesterol diet, and drugs such as cholesterol-lowering drugs are effective. Its use is extremely limited because of its side effects such as hepatic dysfunction.

Chitosan, phytosterol, inositol, and pectin are known substances that lower blood cholesterol in the human body, but the effects or metabolic mechanisms are not clear when phytosterol is excluded. . Phytosterol, a plant sterol due to its structural similarity to cholesterol, has already been identified as a mechanism for inhibiting metabolism of cholesterol absorption in the body through competition with low-density lipoprotein-cholesterol, which is harmful to the human body. As a food additive, the US Food and Drug Administration (FDA) Has been approved.

The cause of the rise of plasma cholesterol is a genetic disease. In this case, the critically ill patient is treated with dietary therapy, cholesterol inhibitors, nicocom, clofibrate, ion exchange resins, and protein steroids. Although drugs are used, these drugs have side effects such as hepatotoxicity, gastrointestinal disorder, carcinogenicity, and the like.

Another major cause of elevated plasma cholesterol is the excessive intake of fat by eating a diet that consumes large amounts of eggs, butter, meat, and the like. In the case of dietary hypercholesterolemia, it usually does not lead to severe hypercholesterolemia, but it is a problem that cholesterol accumulates slowly in the blood vessels from a young age and causes arteriosclerosis in adults, and hypertriglyceridemia (hypertriglyceridemia) In addition, there is a risk of myocardial infarction or cerebral infarction.

The pathological model of experimental hyperlipidemia is divided into exogenous model and endogenous model. Exogenous hyperlipidemic condition models include hypercholesterolemia-induced models caused by high cholesterol dietary loads, that is, hyperlipidemia by administration of vitamin D, cholesterol, olive oil, corn oil, and the like. Models by administration of WR-1339 (Triton WR-1339) (Ham IH et al ., Kor . J. Herbology , 20, pp 45-52, 2005). Among them, Poloxamer 407 is an anesthetic emulsifier (Lee HH et al ., Korean J. Anesthesiol , 40, pp515-521, 2001), poorly soluble drug solubilizer (Choi JY et al ., J. Kor . Pharm . Sci ., 18, pp240-241, 1988), skin ointment (Gill HJ et al ., J. Kor . Pharm . Sci ., 24, ppS27-S32, 1994; Kim KK et al ., J. Kor . Pharm . Sci ., 28, pp15-23, 1998) and the like, but when administered in the human body increases lipids, in particular, triglycerides increase, causing hyperlipidemia. Free fatty acids are produced in adipocytes for at least 24 hours after poloxamer injection (Nash VJ et al ., Pharmacotherapy , 16, pp10-15, 1996) and interfere with the action of lipoprotein lipase (LPL). Decreases the rate of triglyceride degradation (Stellato C. et al ., Br. J. Anaesth ., 67, pp751-758, 1991), and increases blood cholesterol and triglycerides in the liver, which is called 3-hydroxy-3 in the liver. This is because the activity of -methyl glutaryl coenzyme A (HMG-CoA; 3-hydroxy-3-methylglutaryl CoA) is decreased (Wout GM et al ., J. Paren . Sci . Technol ., 46, pp192-200 , 1992).

Alder tree ( Albizzia julibrissin DURAZZ.) is a deciduous arborescent belonging to the legumes (Leguminosae), also called silk tree or mimosa. It is a plant that grows in Japan, China, India, and South Korea, Jeju, southern and central regions, that is, south of the Hwanghae Island (Kim Tae-jung, Korea's Resource Plant II, p194, 1996). It is a small hemisphere and spreads out flat. The gray brown shell and leaves are revived, the superior two times the upper bilobal, and the pink flowers bloom. The flowers of the silk tree have been used in the folklore for the treatment of insomnia, forgetfulness, sore throat and bruises (Kang TH et al., J. Ethnopharmacol ., 71, pp321-323, 2000). As a component of silk tree from the leaf and root bark of potassium-glucopyranosyl-11-hydroxy Silja Sumo carbonate (potassium-glucopyranosyl-11-hydroxyjasmonate ), cis-para-coumarone to one mug animation (p ci s- -coumaroylmagmation) , Quercitrin, afzelin, euscaphic acid ester glucoside, luteolin-7- O- neohesperidoside and (- ) -Syringe resinol [(-)-syringaresinol] and the like (Ueda M and Yamamura S, Tetrahedron Lett . , 40 (44), pp7823-7826, 1999; Jang KG et al., Kor . J. Pharmacogn ., 33, pp18-20, 2002; ... Go JJ et al, Kor J. Pharmacogn, 35, pp194-198, 2004), the flower is a flavonoid glycoside of isopropyl quercitrin (isoquercitrin) was isolated.

Hwanpi ( Albizzia) julibrissin cortex of the Albizia ( Albizzia) julibrissin DURAZZ.) is a stem bark, which has the efficacy of hauls, septic blood, and small sarcoma, and is used for the treatment of ventricular anxiety, melancholia, carbuncles, narcissus and musculoskeletal fractures. JJ et al., Kor . J. Pharmacogn ., 35, pp 194-198, 2004).

The components of the compound skin include: sophoflavescenol, daidzein, geraldone, kuraridin, alpha-spinasterol glucoside, ( E ) -4 -Hydroxyl-dodec-2-enedioic acid [( E ) -4-hydroxy-dodec-2-enedioic acid], kurarinone, isookanin, sulfuretin, luteolin (luteolin), kuraridinol, 3 ', 4', 7-trihydroxyflavone (3 ', 4', 7-trihydroxyflavone), kurarinol, (-)-syringerezinol- 4-O-beta-di-glucopyranoside [(-)-syringaresinol-4- O- β-D-glucopyranoside], soya-cerebroside I, ononin, ikari Inc. lead E ₅ (icariside E _5), Albi debris Sino side a (albibrissinoside a), Albi debris Sino side B (albibrissinoside B) and the saponin compounds line rib side a ₁ (julibroside a _1), line rib side a ₂ ( julibroside A _2), Rib side A ₃ (julibroside A _3), line rib side I (julibroside I), line rib side II (julibroside II), line rib side III (julibroside III) and the line rib side C ₁ (julibrloside C ₁₎ was separated , Phenolic compound 3,4,5-trimethoxylphenol 1- O -beta-D-apiopuranosyl (1 → 2) -beta-D-glucopyranoside [3,4,5-trimethoxyphenol 1 - O -β-D-apiofuranosyl- ( 1 → 2) -β-D-glucopyranoside], Bomi polyol 3'- O - beta -D- Oh Pio furanyl nosil - (1 → 6) - beta -D- gluconic nose Lanoside [vomifoliol 3′- O- β-D-apiofuranosyl- (1 → 6) -β-D-glucopyranoside, (+)-lioniniresinol-9′- O -beta-D-glucopyranosyl- (1 → 4) -beta-D-glucopyranoside [(+)-lyoniresinol-9′- O- β-D-glucopyranosyl- (1 → 4) -β-D-glucopyranoside and (+)-lioniniresinol- 4,9′-di- O- beta-D-glucopyranoside [(+)-lyoniresinol-4,9′-di- O- β-D-glucopyranoside was isolated.

Studies on the leaves of Albizia julibrissin have sedative effects, and studies on the physiological activities of hailwood have reported reports of cytotoxic effects, antioxidant activity and effects on cell cycle inhibition of Jurkat T cells, a leukemia cell line (Jung MJ et al., Arch. Pharm . Res., 26 (3), pp207-209, 2003; Jung MJ et al., Arch. Pharm . Res., 26 (6), pp458-462, 2003; Jung MJ et al., Chem . Pharm . Bull., 52 (12), pp 1501-1503, 2004; Jung MJ et al., Arch. Pharm . Res., 27 (6), pp593-599, 2004; Hiroyuki H et al. , Chem Pharm Bull, 40, pp534-535 , 1992;... Kang TH et al, J. Ethnopharmacol, 71 (1-2), pp321-323, 2000;.... Go JJ et al, Kor J. Pharmacogn, 35, pp194-198, 2004; . Ueda M and Yamamura S, Tetrahedron Lett, 40 (44), pp7823-7826, 1999;.... Jang KG et al, Kor J. Pharmacogn, 33, pp18-20 , 2002; Ikeda T et al., J. Nat. Prod., 60 (2), pp 102-107, 1997; Kinjo J et al., Chem . Pharm . Bull., 40 (12), pp3269-3273, 1992 Ikeda T. et al., Tetrahedron Le tters, 36 (9), pp1509-1510 , 1995; Ikeda T. et al, Bull, Chem Soc Jpn, 68, pp3483-3490, 1995;...... Hwang SG et al, Kor J. Pharmacogn, 33 , pp 29-34, 2002; Hwang SG et al., Yakhak Hoeji , 45, pp 730-738, 2001). However, there has been no teaching or disclosure about the improvement effect of Hailhwanpi extract on hyperlipidemia.

Therefore, the present inventors have completed the present invention by identifying and improving the effect on hyperlipidemia after separating and identifying a novel extract and a lavandulyl flavonoid compound that improve hyperlipidemia in the combined blood.

It is an object of the present invention to provide a pharmaceutical composition useful for the prevention and treatment of hyperlipidemia, which contains an extract of hwanhwanpi with excellent hyperlipidemic activity or kuraridinol isolated therefrom as an active ingredient.

In order to achieve the above object, the present invention is a combination ring julibrissin Cortex) extract or a pharmaceutical composition for the prevention and treatment of hyperlipidemia comprising kuraridinol represented by the following structural formula (1) isolated from it as an active ingredient, and containing a pharmaceutically acceptable carrier or excipient do.

(1)

(One)

The extract is a crude extract or a non-polar solvent soluble extract, the crude extract comprises a lower alcohol such as water, ethanol, methanol, butanol or a mixture thereof, preferably an extract soluble in ethanol, the non-polar solvent soluble extract is dichloro Non-polar solvents selected from methane, chloroform or ethyl acetate, preferably including extracts soluble in ethyl acetate.

The pharmaceutical composition for the treatment and prevention of hyperlipidemia containing the haphwanpi extract and the compound of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition.

Hereinafter, the present invention will be described in detail.

Compound ring extract of the present invention, a non-polar solvent soluble extract and the compound of the flavonoids separated therefrom can be obtained as follows.

After the lyophilized crude extract of the present invention is lyophilized and pulverized and pulverized, the volume of water, methanol, ethanol and about 3 to 20 times, preferably about 3 to 6 times the dry weight of the combined ring skin Lower alcohols such as butanol or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably about 0.5 hour to 2 at an extraction temperature of 20 to 100 ° C., preferably 20 to 80 ° C., with ethanol One, preferably 1 to 5 days by extraction method such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for 1 to 5 times, preferably 3 times continuous extraction, and then filtered under reduced pressure and filtrate The vacuum rotary concentrator was concentrated under reduced pressure at 20 to 100 ° C., preferably 40 to 70 ° C., to obtain a crude ring extract extracted in water, lower alcohol, or a mixed solvent thereof.

Suspension of the crude ring skin extract in water, and then extracted with a solvent in the order of dichloromethane, ethyl acetate, n -butanol to obtain a monocyclic soluble non-polar solvent soluble extract of the present invention, preferably ethyl acetate soluble fraction More specifically, dichloromethane can be obtained by adding dichloromethane to crude chlorophyll extract, that is, ethanol extract of chlorophyllum, to obtain a dichloromethane fraction and a water-soluble fraction.In addition, ethylacetate is added to the water-soluble fraction to give ethyl acetate soluble fraction and water Soluble fractions can be obtained, and finally the water soluble fraction can be extracted with butanol to obtain butanol soluble fraction and water soluble fraction.

For the non-polar solvent soluble fraction, the ethyl acetate soluble fraction is a developing solvent, a mixed solvent of dichloromethane: methanol, preferably a mixed solvent of 20: 1 of dichloromethane: methanol, at a constant volume per hour, preferably 1000 ml. Column chromatography using at speed can be performed to obtain 10 fractions from F1 to F10. The F7 fraction of the fractions was fractionated by 1000 ml per hour using Sephadex LH-20 (Sephadex LH-20, 25-100 μm, Sigma, USA) to separate the compounds of the flavonoids represented by the formula (1). And identifiable.

The present invention provides a method for separating and purifying a crude extract, a nonpolar solvent soluble extract, and a flavonoid compound separated therefrom from a compounded skin.

The antihyperlipidemic effect of the Hwanhwanpi extract obtained by the above process or the flavonoid compounds isolated therefrom in rats induced with hyperlipidemia with poloxamer 407 was significantly lowered because the serum total cholesterol and triglyceride content were significantly lowered. It was confirmed that it can be usefully used as a preventive and therapeutic pharmaceutical composition.

Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of hyperlipidemia, which comprises a haphwanpi extract obtained by the above method and a compound separated therefrom as an active ingredient, and contains a pharmaceutically acceptable carrier or excipient. .

Hwanhwanpi extract of the present invention or a compound isolated therefrom has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of prevention.

The amount and method of application of the pharmaceutical composition comprising the haphwanpi extract of the present invention and a compound separated therefrom may vary depending on the formulation and the purpose of use.

In addition, the composition containing the hwanhwanpi extract of the present invention and a compound isolated therefrom may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Examples of carriers, excipients and diluents that may be included in the composition comprising the Hwanhwanpi extract of the present invention and the compounds separated therefrom 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 Can be.

The composition comprising the Hwanhwanpi extract according to the present invention and the compound isolated therefrom are oral formulations, external preparations, suppositories, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to conventional methods, respectively. It can be formulated and used in the form of sterile injectable solutions.

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid form preparations include at least one excipient such as starch, calcium carbonate, sucrose ( Prepare by mixing sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester 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.

The amount of the Hilhwanpi extract of the present invention and the compound separated therefrom may vary depending on the age, sex, and weight of the patient, but the amount of 0.1 to 100 mg / kg may be administered once to several times daily. In addition, the dosage of the extract and the compound may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

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

Hereinafter, the present invention will be described in detail by Examples, Formulation Examples and Experimental Examples.

However, the following Examples, Formulation Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples, Formulation Examples and Experimental Examples.

Example  One. Blood Crude extract  Produce

Haphwanpi used in the present invention was purchased from Busan medicinal herb. After lyophilization, 15 ℓ of ethanol was added to 3 kg of the powder obtained by lyophilization, and reflux-cooled was extracted three times at regular intervals (12h, 6h, 3h) at 70 ° C., followed by decompression using a filter paper (Watman, USA). After the filtration, the obtained extract was applied to a vacuum rotary concentrator to remove ethanol at 40 ° C., and then 415 g of crude blood extract was obtained as an extracted residue, of which 5 g was used as a sample for animal experiments.

Example  2. Blood  Preparation of Dichloromethane Soluble Extract

Water was added to the crude extract obtained in Example 1 to obtain a water-soluble fraction. 2 l of dichloromethane was added to the water-soluble fraction, followed by mixing three to four times to obtain 2 L of a water-soluble fraction and 2 L of a dichloromethane-soluble fraction. The dichloromethane-soluble fraction was dried to give 72 g of a dichloromethane soluble extract. Obtained.

Example  3. Blood  Preparation of ethyl acetate soluble extract

2 L of ethyl acetate was added to 2 L of the water-soluble fraction obtained in Example 2, followed by mixing 3 to 4 times to obtain 2 L of the water-soluble fraction and 2 L of the ethyl acetate-soluble fraction, and then the ethyl acetate fraction was dried. 24 g of ethyl acetate soluble extract was obtained and used as a sample, of which 5 g was used as a sample for animal experiments.

Example  4. Blood Butanol  Preparation of Soluble Extracts

2 L of butanol was added to 2 L of the water-soluble fraction layer obtained in Example 3, followed by mixing 3-4 times to obtain 2 L of water-soluble fraction and 2 L of butanol-soluble fraction, and then dried butanol-soluble fraction to soluble butanol. 52 g of the extract was obtained and used as a sample, of which 5 g was used as a sample for animal experiments.

Example  5. Blood Soluble extract  Produce

2 L of the water-soluble fraction obtained in Example 4 was dried to obtain 257 g of an aqueous extract, which was used as a sample, of which 5 g was used as a sample for animal experiments.

Example  6. Flavonoids  Isolation of the compound

24 g of the ethyl acetate extract of Example 3 was applied to silica gel column chromatography (SiO ₂ column chromatography, Merck, Germany). At this time, a column chromatography was performed by adding a mixed solvent of dichloromethane: methanol at a rate of 1000 ml / hour as a developing solvent, and 10 fractions were obtained from F1 to F10. F7 fractions of the fractions were applied to Sephadex LH-20 (Sephadex LH-20, 25-100 μm, Sigma, USA), methanol was used as the developing solvent, and fractions were performed at 1000 ml / hour, and 5 ( After dividing into the lower fractions of F7①-F7⑤), the antioxidant activity of each fraction was measured, and the most active F7③ fractions were recrystallized to obtain 150 mg of amorphous orange powdery kuraridinol having the following physical properties. Separated.

Form of material: amorphous orange powder.

Molecular Formula: C ₂₆ H ₃₂ O ₇

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 14.86 (1H, s, OH-9), 7.94 (1H, d, J = 15.6 Hz, H-2), 7.86 (1H, d, J = 15.6 Hz, H-3), 7.43 (1H, d, J = 8.6 Hz, H-6 '), 6.38 (1H, d, J = 2.2 Hz, H-3'), 6.32 (1H, dd, J = 2.2, 8.6 Hz, H-5 '), 6.04 (1H, s, H-6), 4.57 (1H, br s, H-9 "a), 4.47 (1H, d, J = 2.2 Hz, H-9 "b), 3.83 (3H, s, OCH3), 2.55 (2H, m, H-1"), 2.35 (1H, m, H-2 "), 1.64 (3H, s, H-10"), 1.35 (2H, m, H-3 ″), 1.19 (2H, m, H-4 ″), 1.02 (3H, s, H-6 ″), 1.01 (3H, s, H-7 ″).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 191.96 (C-4), 165.28 (C-9), 162.75 (C-7), 161.14 (C-4 '), 160.27 (C-5) , 159.02 (C-2 '), 148.00 (C-8 "), 138.63 (C-2), 130.31 (C-6'), 122.79 (C-3), 113.74 (C-1 '), 110.92 (C -9 "), 108.05 (C-5 '), 106.73 (C-8), 104.41 (C-10), 102.58 (C-3'), 90.66 (C-6), 68.64 (C-5"), 55.49 (OCH ₃ ), 46.41 (C-2 "), 41.57 (C-4"), 29.55 (C-6 "), 29.00 (C-7"), 27.10 (C-1 "), 26.65 (C- 3 "), 17.65 (C-10").

Experimental Example  One. Poloxamer  407 ( Poloxamer  Hyperlipidemia induced by 407) Condition On the rat  About Blood  ethanol Crude extract  Hyperlipidemic effect

In order to confirm the hyperlipidemic effect of polycyclic ethanol crude extract on hyperlipidemic rats induced by Poloxamer 407 (Sigma Chemical, USA), Sprague-Dawley male rats (samples) of about 120 g body weight were tested. Taco, Korea) was used after being adapted to the laboratory environment (temperature 22 ± 2 ℃, humidity 55 ± 5%) for more than two weeks while supplying solid feed (Samyang Blended Feed, Korea) and water. Rats that reached about 200 g were treated with a single intraperitoneal injection of 400 mg / kg of hyperlipidemia-producing substance Poloxamer 407 in 1 ml doses of physiological saline. Two hours after the intraperitoneal administration of poloxamer 407, the samples were initially orally administered and three times were orally administered 24 hours apart. As a sample, 250 and 500 mg / kg of ethanol crude extracts obtained in Example 1 were dissolved in physiological saline containing 5% ethanol and orally administered in a 1 ml dose, and a positive control group lovastatin (lovastatin) , Korea) were 70 and 35 mg / kg, respectively, and the negative control group was orally administered 1 ml of physiological saline containing 5% ethanol, respectively. Blood was collected 12 hours after the last oral administration. The collected blood was left at 25 ° C for 30 minutes and centrifuged to use the supernatant as a sample. The total cholesterol and triglyceride content of the sample was measured using a reagent kit (cholesterzyme-V, triglyzyme-V, Shinyang Chemical, Korea). For total cholesterol and neutral lipid measurement, 10 μl of the sample and 1.5 ml of enzyme solution were mixed well, warmed for 5 minutes in a 37 ° C. hot water bath, and absorbance was measured at 505 nm with the control of the reagent blind within 60 minutes. In this case, the standard and reagent blanks were replaced with the standard solution and the distilled water instead of the sample. The hyperlipidemic effect was obtained by calculating the concentration of cholesterol and triglyceride by the following Equation 1, and expressed as the mean ± standard deviation. SPSS / 10.1 was used for significance test. The significance level was 0.05 (*). The hyperlipidemia group was compared with the normal group (a), and the rest group was compared with the hyperlipidemia group (b) to verify the significance.

Concentration of Standard Solution / Absorbance of Standard Solution = Factor

Factor × absorbance of the sample = improvement in hyperlipidemia (%)

sample Total Cholesterol (mg / ㎗) Neutral Lipid (mg / ㎗)   Normal 95.0 ± 3.9 (34) 77.9 ± 2.8 (25)   Hyperlipidemia group (physiological saline containing 5% ethanol) 279.6 ± 7.9 ^{* a} (100) 295.6 ± 7.0 ^{* a} (100)   250 mg / kg crude ethanol extract 220.9 ± 5.0 ^{* b} (79) 242.4 ± 5.3 ^{* b} (82)   500 mg / kg ethanol crude extract 187.3 ± 2.8 ^{* b} (67) 221.7 ± 2.3 ^{* b} (75)   Lovastatin 35 mg / kg 223.7 ± 4.4 ^{* b} (80) 233.5 ± 5.8 ^{* b} (79)   Lovastatin 70 mg / kg 181.7 ± 6.8 ^{* b} (65) 195.1 ± 5.3 ^{* b} (66)

As shown in Table 1, the total cholesterol and triglyceride concentrations of 500 mg / kg of ethanol crude ethanol crude extract showed an improvement of 33 and 25%, respectively, which was 35 mg / kg of lovastatin (20). , 21%). This indicates that the combined extract of ethanol crude ethanol contains a large amount of excellent cholesterol and triglyceride-lowering substances.

Experimental Example  2. Poloxamer  407 ( Poloxamer  Hyperlipidemia induced by 407) Condition On the rat  About Blood  Nonpolar solvent Fraction  Hyperlipidemic effect

In order to confirm the effect of improving hyperlipidemia of the polycyclic non-polar solvent fractions obtained in Examples 3, 4, and 5 on the hyperlipidemic rats, the results were performed in the same manner as in Experimental Example 1, and the results are shown in Table 2 below. .

As shown in Table 2 below, the total cholesterol and triglyceride concentrations in the butanol soluble fraction 100 mg / kg showed an improvement of 31 and 27%, respectively, which was 35 mg / kg (20, 21%) of the positive control lovastatin. It showed a much more significant effect. Total cholesterol and triglyceride concentrations were improved by 25 and 24% at 100 mg / kg of ethyl acetate soluble fraction, respectively, which was better than the lovastatin 35 mg / kg (20, 21%). 50 mg / kg of ethyl acetate soluble fraction showed 13 and 14% inhibition of total cholesterol and triglyceride levels, respectively.

sample Total Cholesterol (mg / ㎗) Neutral lipid (mg / ㎗)   Normal 86.2 ± 3.9 (34) 72.3 ± 2.0 (25)   Hyperlipidemia group (physiological saline containing 5% ethanol) 253.4 ± 9.9 ^{* a} (100) 289.3 ± 14.5 ^{* a} (100)   50 mg / kg of ethyl acetate soluble fraction 221.6 ± 6.9 ^{* b} (87) 250.7 ± 13.2 ^{* b} (86)   100 mg / kg of ethyl acetate soluble fraction 192.3 ± 9.5 ^{* b} (75) 221.5 ± 11.6 ^{* b} (76)   100 mg / kg of combined skin butanol soluble fraction 174.9 ± 10.1 ^{* b} (69) 211.2 ± 6.9 ^{* b} (73)   100 mg / kg of mixed skin water soluble fraction 230.4 ± 13.6 ^b (90) 265.1 ± 13.9 ^b (91)   Lovastatin 35 mg / kg 202.7 ± 4.4 ^{* b} (80) 228.5 ± 3.8 ^{* b} (79)   Lovastatin 70 mg / kg 164.7 ± 5.8 ^{* b} (65) 190.9 ± 5.3 ^{* b} (66)

Experimental Example  3. Poloxamer  407 ( Poloxamer  Hyperlipidemia induced by 407) Condition On the rat  About Blood  Ethyl Acetate Soluble In fractions  Separated Kuraridinol  ( Kuraridinol Improvement of hyperlipidemia

In order to confirm the hyperlipidemic effect of the compound 1 obtained in Example 6 on the hyperlipidemic condition-induced hyperlipidemic rats, the same procedure as in Experimental Example 1 was carried out. Shown in

As shown in Table 3, the 20 mg / kg treatment group of the flavonoid compound Kuraridinol (Kuraridinol, Compound 1) compared with the negative control group had a total cholesterol and neutral lipid concentration of 44 and 24%, respectively. The effect was improved, which was much more effective than the lovastatin 35 mg / kg treated group (20, 21%). Kuraridinol has been shown to improve hyperlipidemia as a single compound.

sample Total Cholesterol (mg / ㎗) Neutral Lipid (mg / ㎗)   Normal 93.8 ± 10.0 (35) 81.2 ± 4.1 (27)   Hyperlipidemia group (physiological saline containing 5% ethanol) 268.0 ± 12.0 ^{* a} (100) 300.9 ± 5.9 ^{* a} (100)   Kuraridinol 10 mg / kg 194.3 ± 6.8 ^{* b} (72) 243.4 ± 2.8 ^{* b} (81)   Kuraridinol 20 mg / kg 149.6 ± 16.3 ^{* b} (56) 228.1 ± 10.9 ^{* b} (76)   Lovastatin 35 mg / kg 214.4 ± 6.5 ^{* b} (80) 237.7 ± 4.0 ^{* b} (79)   Lovastatin 70 mg / kg 176.9 ± 5.4 ^{* b} (66) 192.6 ± 8.8 ^{* b} (64)

Hereinafter, the preparation example of the pharmaceutical composition will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Formulation example  One. Powder  Produce

Curaridinol 20 mg of Example 6

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 Tablets

Curaridinol 10 mg of Example 6

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. Manufacture of capsule

Curaridinol 10 mg of Example 6

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 Injectables

Curaridinol 10 mg of Example 6

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. Liquid  Produce

Curaridinol 20 mg of Example 6

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 and dissolved in purified water, lemon flavor is added thereto, the above components are mixed, and then, purified water is added to adjust the total volume to 100 ml, and then sterilized by filling into a brown bottle. do.

Hwanhwanpi of the present invention ( Albzzia julibrissin Cortex) extract or kuraridinol compound isolated therefrom shows an improvement effect on hyperlipidemia closely related to atherosclerosis, and thus can be used as a pharmaceutical composition for the prevention and treatment of hyperlipidemia and atherosclerosis.

Claims (5)

Hwanhwanpi ( Albzzia julibrissin cortex) or a pharmaceutical composition for the prevention and treatment of hyperlipidemia comprising kuraridinol represented by the following structural formula (1) isolated therefrom as an active ingredient, and containing a pharmaceutically acceptable carrier or excipient:

(One) The pharmaceutical composition according to claim 1, wherein the extract is a crude extract or a nonpolar solvent soluble extract. The pharmaceutical composition according to claim 2, wherein the crude extract is an extract available in water, ethanol, methanol, butanol lower alcohol or a mixed solvent thereof. The pharmaceutical composition according to claim 2, wherein the nonpolar solvent soluble extract is an extract soluble in a nonpolar solvent selected from dichloromethane, chloroform or ethyl acetate. According to claim 1, wherein the extract or compound is a pharmaceutical composition, characterized in that contained in 0.1 to 50% by weight relative to the total weight of the composition.