KR20020085507A - Preparation of 3,4-dicaffeoylquinic acid and use thereof as anti-cholesterol agent - Google Patents

Abstract

PURPOSE: A process of preparing 3,4-dicaffeoylquinic acid having a cholesterol-lowering effect from a Ligularia fischeri extract is provided. Whereby, the compound is effectively used in a pharmaceutical composition for treatment of ailments such as high cholesterol levels or a food composition for reducing the high cholesterol levels. CONSTITUTION: Ligularia fischeri var. spiciformis is extracted in methanol, suspended or dissolved in water and then extracted in ether. The obtained extract contains 30% by weight of 3,4-dicaffeoylquinic acid or 5-0-butyl-3,4-dicaffeoylquinic acid having a cholesterol-lowering effect as an effective ingredient and a composition containing the extract can be used for food, beverage, medicine, tea or other health auxiliary foodstuffs.

Description

Preparation method of 3,4-dicafeoylquinine acid and use as cholesterol lowering agent {Preparation of 3,4-dicaffeoylquinic acid and use thereof as anti-cholesterol agent}

Cholesterol is a constituent of biological membranes and one of the essential nutrients for the human body as one of the starting materials for hormone synthesis. However, when cholesterol is present in excessive concentrations in blood, it can cause various diseases such as atherosclerosis, so it is desirable to maintain it at an appropriate concentration. In order to control blood cholesterol levels, low-cholesterol diets and drugs such as cholesterol-lowering drugs have been developed, but dietary therapy is limited to prevention, and in the latter case, side effects such as liver dysfunction have been reported. Because of this limitation, the development of safer and more effective cholesterol-lowering substances is urgently needed.

In the past, pure compounds have been preferred as raw materials for pharmaceuticals. In recent years, however, natural extracts containing various substances have been developed as additives for functional foods or as raw materials for pharmaceuticals, because a mixture containing several substances rather than a single substance exhibits excellent physiological activity. This is because many natural extracts have been proved to be harmless to humans over a long period of time. In view of this, many natural extracts with low toxicity and excellent physiological effects have been developed in Korea, such as cinnamon oil, gamma-orizanol, horse chestnut extract, silymarin (a mixture of silybin and silycristine), and garlic oil.

Overflowing is one of the wild vegetables edible as an asteraceae plant native to Korea, whose scientific name is Ligularia fischeri var. Spiciformis . From the plant of the genus Ligularia (Zhao Y. et al., Journal of Natural Products. 1994; 57: 1626-1630; Gao K. et al., Planta Medica 1997; 63: 461-463, Jia Z. et al., Journal of Natural Products 1993; 56: 194-499; Bohlmann F. and, Fritz U., Phytochemistry 1980; 19: 2471-2472), phenylpropanoid system (Zhao Y. et al., Phytochemistry 1994; 37: 1149-1152; Ma B. et al., Phytochemistry 1997; 46: 915-919), pyrrolidinine type alkaloids (Asada Y. and Furuya T., Chemical and Pharmaceutical Bulletin 1984; 32: 475- 482), and the most known plant components are eremophyllan sesquiterpenes (Park HJ et al., Planta Med. 2000; 66: 783-784).

The inventors of the present inventors studied the overflow, and the solvent extract fraction contained a large amount of a series of quinic acid derivatives, and the effect of these solvent extracts and the purified 3,4-dicapoylquininic acid lowered serum serum cholesterol Discovered excellent and led to the present invention.

The present invention provides a flushing extract containing a large amount of quinic acid derivatives and having a lowering effect on cholesterol, and a method of preparing the overflowing extract.

The present invention provides a method for preparing 3,4-dicaffeoylquinic acid (3,4-DCQA) from an overflow or overflow extract.

The present invention provides a composition for inhibiting cholesterol having the overflow extract or 3,4-DCQA as an active ingredient. Examples of such compositions include foods, beverages, pharmaceuticals, teas and other health supplements.

The present invention provides an overflow extract having a cholesterol-lowering effect containing a large amount of quinic acid derivatives and a method of preparing the same. Overflow is a Korean specialty plant whose name is Ligularia fischeri var. Spiciformis and is distributed mainly in Gangwon-do. Azalea is a bicotyledonous plant of the Asteraceae family. It is a perennial wild vegetable with the development of a latex leaf, and the related plants include bear, gondalbi, long-leaf bear, hairy duck, horned bear, royal bear, giant bear, and dog tobacco. Overflow is plant ecologically similar to Ligularia fischeri Turcz , one of the more widely used wild vegetables. In China, the bear odor is called horochil, and it is used as a herbal medicine for bruises, back pain, Jinhae, expectorants. The bear odor contains a lot of sesquiterpene-based compounds, and the main pharmacological components of the bear odor are chamomile, jacobine, and amellene. It is reported that antimutagenic activity is shown (Seo Jongtaek, Folk Vegetables (wild vegetables), Nonghyup, pp. 35-44 (1997)).

In Korea, the bear odor has been used for jaundice, solitary, arthritis, hemorrhoids and liver disease. Bear smell and overflow are grown in Daegwallyeong area of Korea. It is a rare plant that grows in Korea. It is a rare botanical resource that can be eaten raw. It grows under deep forests, wet and fertile vegetation and valleys.

In order to prepare the overflow extract of the present invention, the overflow may be used by drying or not dried. It is easy to use a dry sample of overflow, and in this case, it is preferable to use the finely ground to facilitate the extraction. To prepare the overflow extract, the overflow or crushed overflow is extracted with an alcohol solvent. The alcohol solvent can be used without limitation the alcohol solvent commonly used in the field of natural product extraction. The alcohol solvent is preferably a lower or intermediate alcohol. Examples of alcohol solvents are methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol, amyl alcohol or allyl alcohol. More preferred solvent is ethyl alcohol or butyl alcohol. The amount of the extraction solvent is sufficient to cover the sample sufficiently. The alcohol solvent is extracted by heating at reflux for about 2 to 10 hours, preferably about 3 to 7 hours. Filtration of the extract or concentration of the filtered solution may optionally include.

In the second step, the alcohol extract is suspended in water and the suspension is extracted with ether. The ether fraction contains sesquiterpenes that exhibit cytotoxicity. Therefore, the ether fractions are collected and discarded and the remaining exports are taken. Ether extraction can be repeated 2-4 times as needed.

In the third step, the remaining water layer after ether extraction is extracted with ethyl acetate to prepare an overflow extract. The ethyl acetate fraction contains at least 30% by weight or more of quinic acid derivatives (Examples 1 and 2).

Further silica gel chromatography of ethyl acetate fractionated overflows yielded three substances that are the main quinic acid derivatives of the fraction, namely chlorogenic acid, 3,4-DCQA and 5-0-butyl-3,4-DCQA. Disconnect. When chloroform-methanol-water (65: 35.10, lower layer) was used as the solvent, chlorogenic acid eluted first, followed by 3,4-DCQA and 5-0-butyl-3,4-DCQA. 3,4-DCQA and 5-0-butyl-3,4-DCQA can be separated using an octadecsilane column (Example 3).

The three major quinic acid derivatives concentrated in the overflow extract fractionated with ethyl acetate were chlorogenic acid, 3,4, respectively, by methods such as melting point, IR-spectrum, IH- and 13C-spectrum, UV-absorbance and mass spectrum. It was identified as -DCQA and 5-0-butyl-3,4-DCQA (Table 1 and Table 2).

It was confirmed through animal experiments that the overflow extract, which was fractionated with ethyl acetate, contained at least 30% of quinic acid derivatives and had an excellent effect of lowering serum cholesterol. Cholesterol-lowering effects were also observed in 3,4-DCQA.

Literature has reported that DCQA and methyl-DCQA act as protective agents of hepatic function as an active ingredient of propolis extracts. (P. Basnet et al., Biol. Pharm. Bull., 19 (4), 655 (1996)). Coffee beans contain a lot of 3,4-, 3,5-, 4,5-di-0-cafeoylquininic acid, and Loch's is 1,3,4 isolated from Pluchea symphytifolia. 1,3-di-0- [3,4-bis- (3,4-dihydroxyphenyl) -cyclobutane-1,2-dicarbonyl] -4,5-di Caffeoylquinic acid has been shown to have antimicrobial activity (E. Scholtz et al., Planta Med., 60, 360 (1994)), and Nishizawa is a 3,4-di- derivative that inhibits lipooxygenase activity. Caffeoyl-5- (3-hydroxy-3-methylglutaroyl) quinic acid has been isolated from Gardeneuiae Fructus (M. Nishizawa and Y. Fujimoto, Vhem. Phar, Bull. , 1419 (1986)). However, the cholesterol lowering effect of ethylacetate fractionated overflow extract and 3,4-DCQA, in which quinic acid derivatives are concentrated, is the first finding by the present inventors.

The present inventors used Sprague-Dawley male rats to demonstrate the therapeutic effect of hypercholesterolemia by the overflow extract or 3,4-DCQA. Rats were fed high fat and serum total cholesterol, HDL-cholesterol and LDL-cholesterol levels were investigated and the atherosclerotic index (A. I.) was calculated.

In general, a high cholesterol animal model does not form well even if a large amount of cholesterol is administered to many experimental animals. However, dietary supplementation with choline acid, one of the bile acids, can dramatically reduce the metabolism of cholesterol into choline acid, making serum high cholesterol model mice. Cholic acid dramatically decreases the activity of 7a-hydroxylase, the first step in cholesterol metabolism, resulting in a significant increase in serum cholesterol.

In this experimental model, as shown in Table 2, serum total cholesterol was increased about 1.8 times and LDL-cholesterol was increased about 3.6 times. HDL-cholesterol did not increase much. The excess ether fraction hardly reduced serum cholesterol, while the ethyl acetate fraction containing quinic acid derivatives and 3,4-DCQA reduced both total cholesterol and LDL-cholesterol levels in a significant range. The cholesterol lowering effect was highest in 3,4-DCQA. This indicates that one of the active ingredients lowering cholesterol is 3,4-DCQA (Table 4). Specifically, the overflow extract and 3,4-DCQA significantly reduced LDL-cholesterol associated with atherosclerosis, while not significantly increasing HDL-cholesterol. Atherosclerosis index (A.I.) was significantly decreased in the group treated with ethyl acetate fraction or 3,4-DCQA.

In atherosclerosis patients, clinically, total cholesterol is generally higher, the amount of LDL-cholesterol is higher than normal, and VLDL-cholesterol is the same type as normal, and VLDL-cholesterol is increased and LDL- There are several types, including normal cholesterol. HDL-cholesterol is high in normal people and low in atherosclerosis patients because HDL-cholesterol plays a role in depositing cholesterol in blood vessels and in liver (Harper's Biochemistry, Robert K Murray et al., Prentice-Hall international Inc., 1993 , pp. 266-278).

Due to the complex metabolism and clinical significance of these lipoprotein cholesterol, the use of LDL-cholesterol / HDL-cholesterol level or arteriosclerosis risk index (AI) -Cholesterol) / HDL-cholesterol}. Recently, the arteriosclerosis risk index (A. I.) has been widely used.

As shown in Table 2, the arteriosclerosis risk index increased about 1.7 times in the high cholesterol diet group than the negative control group. The excess methane extract and ethyl acetate fraction reduced the risk of atherosclerosis. The oral administration of 250 mg / kg and 500 mg / kg of methane extract reduced the atherosclerotic risk index to 34% and 66%, respectively. The ether and butanol fractions were not very beneficial in improving the atherosclerotic risk index. This result is supported by the fact that the ether fraction contains cytotoxic substances, which may lead to lipid peroxidation.

Although 10 mg / kg of 3,4-DCQA was intraperitoneally administered, the effect of reducing arteriosclerosis risk index was only 11%. It was. This suggests that the ethyl acetate fraction containing quinic acid derivatives and 3,4-DCQA have an effect of reducing serum cholesterol, and in particular, 3,4-DCQA is a very good substance for lowering cholesterol. Changes in total cholesterol and LDL-cholesterol were not significantly different from those in the atherosclerotic risk index. There was no significant effect on the amount of HDL-cholesterol throughout the entire treatment group.

New drugs related to cholesterol lowering include cholestyramine resins that inhibit the intestinal-to-hepatic circulation of bile acids, β-sitosterol, a phytosterol that antagonizes the absorption and distribution of cholesterol, and HMG-CoA reductase that acts on the biosynthesis of cholesterol ( The reductase inhibitors mevastatin and lovastatin are well known, and probucol is known to be applied to atherosclerosis patients by increasing LDL metabolism. Probucol has been used as a drug for atherosclerosis, but the mechanism is unknown. However, it has been established that the recent antioxidant action on probucol is related to LDL metabolism (Harper's Biochemistry, Robert K Murray et al., Prentice-Hall international Inc., 1993, pp. 266-278). DCQA and ethyl acetate fractions revealed in the present invention showed an antioxidant effect in a separate experiment.

The present invention provides a composition having a lowering effect on the cholesterol containing the ethyl acetate fraction or 3,4-DCQA. The present invention provides a pharmaceutical composition for lowering cholesterol comprising the overflow extract or 3,4-DCQA in an amount sufficient to treat hypercholesterolosis and a method for lowering cholesterol by administering the same. The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier. Carriers include solvents, dispersion media, absorption retardants, and the like commonly used in the pharmaceutical art. The pharmaceutical composition of the present invention may be administered via any general route as long as it can reach the desired tissue. Therefore, the composition of the present invention can be administered topically, orally, parenterally, intravenously, intramuscularly, subcutaneously, transdermally, and can be formulated into solutions, suspensions, tablets, pills, capsules, sustained release preparations and the like. Dosage is determined in consideration of the skill of the art depending on the type and extent of the disease, age, sex, etc. of the patient.

The pharmaceutical composition for lowering cholesterol according to the present invention contains an overflow extract or 3,4-DCQA as an active ingredient. Pharmaceutically acceptable carriers that are compatible with different formulations include all forms of diluents or solvents, fillers, scattering agents, binders, dispersants, disintegrants, surfactants, lubricants, excipients, and wetting agents. Furthermore, if necessary, commercially available dissolution aids, buffers, preservatives, colorants, flavors, sweeteners, and the like may also be added to the drug.

The various forms of medicaments described above can be prepared by conventional methods, for example in the preparation of oral dosage forms such as tablets, capsules, granules and pills, they are excipients such as white sugar, lactose, glucose, starch, mannitol ; Binders such as syrup, gum arabic, sorbitol, trigacanth rubber, methylcellulose, polyvinylpyrrolidone and the like; Disintegrants such as starch, carboxymethyl cellulose and its calcium salts, microcrystalline cellulose, polyethylene glycol and the like; Lubricants such as talc, magnesium stearate, calcium stearate, silica, and the like; It may be prepared by conventional methods using wetting agents such as sodium laurate, glycerol and the like.

In the preparation of injectables, solutions, emulsions, suspensions and syrups, they are prepared by conventional methods, such as ethyl alcohol, isopropyl alcohol, propylene glycol, 1,3-butylene glycol, polyethylene glycol, castor oil and the like. Solvent for component dissolution; Sorbitol fatty acid ester, polyoxyethylene sorbitol fatty acid ester, polyoxyethylene ester, hydrogenated castor oil

Surfactants such as polyoxyethylene ether, lecithin and the like; Manufactured using appropriate preservatives such as cellulose derivatives such as sodium carboxymethyl cellulose, methyl cellulose, suspending agents such as natural rubbers such as trigacanth rubber, gum arabic, esters of paraoxybenzoic acid, benzalkonium chloride, sorbitan fatty acid salts, etc. Can be. Cytotoxicity was not observed when the present invention overflow or 3,4-DCQA was administered.

The present invention provides a food composition containing overflow extract or 3,4-DCQA. The food used as a base material is not specifically limited, Various foods, such as water, soft drinks, fruit drinks, confectionery, discharge, noodles, seasonings, etc. can be based on. The food of this invention is easily made by adding the process of adding the composition of this invention mentioned above to the manufacturing process of the base food, or by adding the process of adding the composition of this invention mentioned above after manufacture of the food of base materials. You can get At this time, you may add a taste and odor correction agent as needed. The content of the above-mentioned composition in the food of the present invention varies depending on the kind, preference, etc. of the food of interest, but preferably contains an amount sufficient to exhibit a lowering effect on the cholesterol contained in the overflow extract or 3,4-DCQA. . Since the food of this invention obtained in this way contains the overflow extract of this invention or 3, 4-DCQA, it can be usefully utilized as a functional food with a cholesterol lowering effect.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the examples.

<Example>

Example 1

Preparation of Overflow Samples and Preparation of Alcohol Extracts

Ligularia fischeri var. Spiciformis , Compositae, grown in Jinbu-myeon, Pyeongchang-gun, Gangwon-do, Korea , was purchased and used as a material. The plants used in the experiments were referred to Dr. Yoo Myung, a biology botanist at Kangwon National University for taxonomic identification. After the overflow was dried in the shade, the dried sample was ground finely and used as an extraction material.

The dry pulverized overflow (dry weight, 2 kg) was placed in a back flask, and methanol as an extraction solvent was added to sufficiently cover the sample. The reflux cooler was mounted and the active ingredient was extracted by heating using a heating mantle for 5 hours. The methane was filtered out using a filter paper (Watman No. 1, Whatman) and the filtrate was concentrated under reduced pressure using a vacuum concentrator while warming the filtrate with a water bath at 40 ° C. As a result, l90 g of methanol extract was obtained.

Example 2

Fraction of extract

170 g of the methanol extract obtained in Example 1 was suspended in 800 ml of distilled water. The solution was placed in a separatory funnel in a 2 L container, 800 ml of ether was added thereto, shaken vigorously for about 5 minutes, and left at room temperature for 10 minutes. The ether layer located in the upper layer was taken and extracted twice more with ether in the same manner. 600 ml of ethyl acetate was added to the aqueous phase and fractionated three times in the same manner as in the ether fraction. The upper ethyl acetate layer was collected and stored in another container. After extracting ethyl acetate, 600 ml of butane was added to the remaining aqueous layer and fractionated three times, and the upper layer of butane was taken. The fraction samples obtained through the fractionation process were separately concentrated in a vacuum concentrator to obtain 25 g of ether fraction, 63 g of ethyl acetate fraction and 45 g of butanol fraction.

Example 3

Silica Gel Chromatography

A column with an inner diameter of 7 cm was fixed vertically to the stand and the bottom part of the column was closed with cotton. 1.2 L of the previously developed developing solvent chloroform-methanol-water (65:35:10, lower layer) and 300 g of silica gel (Kiesel gel 60 (1.07734.1000), Merk) were suspended in an Erlenmeyer flask, and then quickly added to the column and The stopper was opened to remove the developing solvent. Using this, all remaining silica gel in the Erlenmeyer flask was washed and filled in the column. Elution was made until the developing solvent was on top of the silica gel and the column was capped.

In order to adsorb the sample onto silica gel, 20 g of the ethyl acetate fraction prepared in Example 2 was mixed well with 200 ml of methanol to dissolve the fraction. Then, 30 g of silica gel was added, and the solvent was evaporated while warming to 60 ° C. in a vacuum concentrator. The dried powdery product obtained was added to mortar and then finely ground the pestle. This was carefully loaded onto a preprepared column using a spoon. Aliquots of 60 ml were continued to obtain 100 fractions altogether.

All the fractions obtained were analyzed for elution by thin layer chromatography (TLC) (TLC plate; Siliga gel 60F ₂₅₄ (1.057l5), Merck). TLC was by the method described in the manufacturer's manual. Spots were confirmed by UV irradiation and ammonia dipping. The white needle formed when the 18th to 22nd fractions were concentrated and dissolved in methanol was identified as chlorogenic acid by spectroscopic analysis (yield amount: 130 mg). Fractions 55 to 90 containing 3,4-DCQA and 5-0-butyl-3,4-dicafeoylquinolinic acid (retention volume, 3.3 L-5.4 L) were combined and concentrated, followed by column chromatography. Separated.

60 g of filler octadecylsilane (ODS) was added to the column with a diameter of 3 cm by mixing with a developing solvent (methanol-water (10: 2)). The solvent was slowly withdrawn and the stopper closed when the developing solvent came to the top of the column stop phase. Samples for separation were dissolved in 20 ml of the same developing solvent and loaded onto the column. While developing the ODS column chromatography, a sufficient amount of developing solvent was injected when the sample solution came to the top of the stationary phase. The eluate was aliquoted 20 ml to obtain 50 fractions. Thirty fractions were TLC in the same manner as above and spots were identified by UV irradiation. From 18 to 31 fractions were concentrated and dissolved in methanol to obtain a white amorphous powder produced by the spectroscopic analysis was confirmed to be 3,4-DCQA (yield 3.2 g). Fractions 38 to 46 were concentrated and left to dissolve in methanol. When the white amorphous powder produced at this time was filtered off, compound 5-0-butyl-3,4-DCQA was obtained (amount 1.2 g).

Example 4

Structure Identification of Separation Compounds

Melting points of the three compounds separated were measured using a melting point measuring instrument (Electrical Digital Melting Point Apparatus, Electrothermal). The luminescence was measured at 25 ° C. using a JASCO DIP-360 Digital Polarimeter (JASCO) by the manufacturer's manual method. IR spectra were measured by a KBr Disk method using a Bomem MB-100 FT-IR Spectrometer, and mass spectra were measured by Finnigan Mat TSQ-700. ¹ H- and ¹³ C-NMR spectra were measured on a Bruker AM-500 Spectrometer with the addition of the internal standard tetramethylsilane (TMS). UN absorbance was measured by UV-160A UN-visible Recording Sepctrophotometer (Shmadzu). At this time, the wavelength and absorbance at the absorption maximum were read, and the log? Value was calculated and recorded. Chlorogenic acid, 3,4-DCQA and 5-0-butyl-DCQA were identified by the above method.

Chlorogenic acid: Molecular formula C ₁₆ H ₁₈ O ₉

Melting point 208 °

[α] _D -35.2 (c = 2.8) at 26 ° C

FAB-MS m / z: 377 [M + Na] ⁺

¹ H-NMR (500 MHz, DMSO): Table 1

¹³ C-NMR (125 MHz, DHSO): Table 2

3,4-DCQA: molecular formula C ₂₅ H ₂₄ O ₁₂

[α] _D -225 ° (MeOH)

FAB-MS m / z: 539 [M + Na] ⁺

¹ H-NMR (500 MHz, DMSO): Table 1

¹³ C-NMR (125 MHz, DMSO): Table 2

5-0-butyl-3,4-DCQA: UV λ _max (MeOH) nm (log ε): 218 (4.30), 242 (4.12), 299 sh (4.16), 330 (4.32)

IR v (cm _-1 ): 3372 (broad, OH), 1701 (α, β -unsaturated ester), 1630, 1602, 1516, 1439 (aromatic C = C), 1285 (C-0)

FAB-MS (m / z): 591 [M + Na] ⁺

¹ H-NMR (500 MHz, DMSO): Table 1

¹³ C-NMR (125 MHz, DMSO): Table 2

Chemical shifts can be interchanged between two caffeyl oils in the same molecule.

^a 5-0-butyl-3,4-di-0-caoylquinic acid

^b 3,4-di-0-cafeoylquinic acid

Chemical shifts can be interchanged between two caffeyl oils in the same molecule.

^a 5-0-butyl-3,4-di-0-caoylquinic acid

^b 3,4-di-0-cafeoylquinic acid

^c Chemical shift values are cited in the following literature. (Arch. Pharm. Res. 14 (2) 114-117 (1991))

Example 5

High Cholesterol Diet

Sprague-Dawley male rats (120 ± 5g) were purchased from the Korean Experimental Animal Development Company and pre-stocked with solid feed for a week. Formulation of the high-fat diet is shown in Table 3. The diet and water were freely ingested for 6 weeks, and the last one week was orally administered with an oral needle zonde extract. The temperature of the feeding room was maintained at 22 ± 2 ℃, the humidity was 55 ± 5%, and the contrast was adjusted every 12 hours.

Example 6

Measurement of Serum Cholesterol and Estimation of Atherosclerosis Index

Total cholesterol, triglyceride and phospholipid amounts were determined by the method of Noma et al. Using a commercial enzyme kit (Sigma) (Noma, A. et al., Clin. Chem ., 24, 1504 (1978); Noma, A. et al, Clin. Chem ., 25, 1480 (1979)). HDL- and LDL-cholesterol amounts were measured according to the manufacturer's manual using a measurement kit (Sigma).

In the high-fat diet rat group, methanol extract, ether fraction, butyl alcohol fraction and ethyl acetate fraction (250, 500 mg / kg) prepared in Examples 1 to 3 were orally administered daily over a week or 3,4- DCQA (10, 20 mg / kg) was intraperitoneally administered. After 24 hours, rats were caught to measure serum cholesterol levels and the arteriosclerosis index was calculated (Table 2). Total cholesterol and LDL-cholesterol contents were significantly increased in the high-fat diet group than the normal group, but there was no significant change in the content of HDL-cholesterol. Total cholesterol and LDL-cholesterol contents were decreased in the high fat diet group compared to the control group, but the HDL-cholesterol content was not significantly changed.

Values are mean ± S.D. (n = 8). 3,4-DCQA = 3,4-dicafeoylquinic acid. Values in parentheses for total cholesterol, LDL, and atherosclerosis index (A. I.) represent the mean values of percent inhibition or percent reduction to normal values, and values in parentheses for HDL represent the mean percentage of gains relative to normal values (%).

Overdose is a plant that has been edible for a long time in Korea and the extract of overdose or 3,4-DCQA isolated from it is excellent in lowering blood cholesterol level and is a pharmaceutical composition for treating hypercholesterolemia or foods for reducing cholesterol It can be usefully used in compositions and the like.

Claims (9)

Overflowing extract, characterized in that it contains more than 30% by weight of the quinic acid derivative, has a cholesterol-lowering effect. The method according to claim 1, wherein the quinic acid derivative is chlorogeninic acid, Extract characterized in that it comprises at least one substance selected from 3,4-dicafeoylquinic acid or 5-0-butyl-3,4-dicafeoylquinic acid. Extracting Ligularia fischeri var. Spciciformis with methanol; Suspending or dissolving the methanol extract in water; Extracting the suspension or melt with ether; Extracting the ether and extracting the remaining aqueous layer with ethyl acetate. A method for separating 3,4-dicafeoylquinic acid from overflow. A method for separating chlorogenic acid from overflow. A pharmaceutical composition for lowering cholesterol, comprising the overflow extract of claim 1 Cholesterol lowering pharmaceutical composition, characterized in that it contains 3,4-dicafeoylquinic acid A food composition for lowering cholesterol, comprising the overflow extract of claim 1 Cholesterol lowering food composition characterized by containing 3,4-dicafeoylquinic acid