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WO2003059369A1 - Extrait d'hovenia dulcis thunb insoluble dans l'alcool inferieur et polysaccharide isole a partir de cet extrait - Google Patents

Extrait d'hovenia dulcis thunb insoluble dans l'alcool inferieur et polysaccharide isole a partir de cet extrait Download PDF

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Publication number
WO2003059369A1
WO2003059369A1 PCT/KR2002/001449 KR0201449W WO03059369A1 WO 2003059369 A1 WO2003059369 A1 WO 2003059369A1 KR 0201449 W KR0201449 W KR 0201449W WO 03059369 A1 WO03059369 A1 WO 03059369A1
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WO
WIPO (PCT)
Prior art keywords
lower alcohol
insoluble fraction
polysaccharide
methanol
hovenia dulcis
Prior art date
Application number
PCT/KR2002/001449
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English (en)
Inventor
Chun Soo Na
Nam Chul Jung
Sea Hyun Kim
Original Assignee
Lifetree Biotech Co., Ltd.
Forestry Research Institute
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR10-2002-0002772A external-priority patent/KR100403721B1/ko
Application filed by Lifetree Biotech Co., Ltd., Forestry Research Institute filed Critical Lifetree Biotech Co., Ltd.
Priority to AU2002367003A priority Critical patent/AU2002367003A1/en
Publication of WO2003059369A1 publication Critical patent/WO2003059369A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/72—Rhamnaceae (Buckthorn family), e.g. buckthorn, chewstick or umbrella-tree
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

Definitions

  • the present invention relates to a lower alcohol-insoluble extract of Hovenia dulcis Thunb, a polysaccharide isolated theref om having hepatoprotective and hangover-resolving activities and a pharmaceutical composition or health care food containing same.
  • Hepatitis has been afflicting an ever-increasing number of the world population, and due to the lack of therapeutically effective drugs, it usually progresses to chronic hepatitis, liver cirrhosis or cancer.
  • Various types of hepatitis may be developed when a patient is exposed to several factors such as stress, excessive consumption of alcohol, and/or hepatotoxic substances.
  • hepatotoxic substances are CCI 4 , D-galactosamine, lipopolysaccharide (LPS), bromobenzene and aldehydes such as acetaldehyde which is known as an intermediate in the metabolic pathway of alcohol. Accordingly, there have been many attempts to find new drugs which can protect the liver from such hepatotoxic substances or restore the liver function damaged thereby.
  • triterpene glycosides isolated from the seed and fruit of Hovenia dulcis Thunb are known to inhibit the release of histamine and the absorption of alcohol in human body (Yoshikawa, K. T. et al., (1995) Chem. Pharm. Bull. Tokyo, 43(3), pp532-534); and the fruit of Hovenia dulcis Thunb has been found to inhibit liver damage caused by carbon tetrachloride or D- galactosamine/ lipopolysaccharide (Hase K. et al., (1997) Chem. Pharm. Bull. Tokyo, 20(4), pp381-385).
  • a lower alcohol-insoluble fraction obtained by treating a hot-water extract of dried Hovenia dulcis Thunb with a lower alcohol.
  • a polysaccharide having a potent hepatoprotective activity which is isolated from said lower alcohol-insoluble fraction.
  • Fig. 1 a schematic procedure for preparing a methanol-insoluble fraction of an extract of the fruit-peduncle of Hovenia dulcis Thunb and a polysaccharide isolated therefrom;
  • Fig. 2 a DSC scan of the methanol-insoluble fraction
  • Fig. 3 a MALLS spectrum of the methanol-insoluble fraction
  • Fig. 4 an IR spectrum of the methanol-insoluble fraction
  • Fig. 5 a UV spectrum of the methanol-insoluble fraction
  • Fig. 6 GC spectrums of the methanol-insoluble fraction
  • Fig. 7 MALLS spectrums of the methanol-insoluble polysaccharide
  • Fig. 8 an IR spectrum of the methanol-insoluble fraction
  • Fig. 9 an NMR spectrum of the methanol-insoluble fraction
  • Fig. 10 in vitro protein synthesis activity of the methanol-insoluble fraction to a carbon tetrachloride-induced liver slice culture
  • Fig. 11 in vitro protein synthesis activity of the methanol-insoluble fraction to a galactosamine LPS-induced liver slice culture
  • Fig. 12 in vitro protein synthesis activity of the methanol-insoluble fraction to a bromobenzene-induced liver slice culture
  • Fig. 13 LDH release inhibitory activity of the methanol-insoluble fraction to the bromobenzene-induced liver slice culture
  • Fig. 14 in vitro protein synthesis activity of the polysaccharide shown in Fig. 7 to the bromobenzene-induced liver slice culture;
  • Fig. 15 activity in lowering alcohol concentration in blood of the methanol-insoluble fraction and the polysaccharide shown in Fig. 7;
  • Fig. 16 alcohol dehydrogenase activity of the methanol-insoluble fraction and the polysaccharide shown in Fig. 7.
  • the lower alcohol-insoluble fraction of a hot-water extract of Hovenia dulcis Thunb of the present invention may be prepared in two steps. First, a hot- water extract of dried Hovenia dulcis Thunb is obtained using a high-pressure extraction procedure and then an insoluble fraction is obtained treating the hot- water extract thus obtained with a lower alcohol.
  • a fruit peduncle of Hovenia dulcis Thunb is sliced and dried in the shade.
  • An appropriate amount of water is added to the dried slices of the Hovenia dulcis Thunb fruit peduncle and the resulting mixture is kept at a temperature ranging from 110 to 150 ° C, preferably 120 to 125 ° C, under a pressure ranging from 1 to 3 arm., preferably 1.5 atm., for a period ranging from 15 minutes to 48 hours, preferably 30 minutes to 12 hours.
  • the mixture is cooled to room temperature and filtered, and the filtrate is lyophilized pursuant to a conventional lyophilizing method, to obtain a hot-water extract.
  • the hot-water extract is further dried at room temperature, evaporated under a reduced pressure e. g., -1.5 atm, and extracted with a lower alcohol, preferably methanol, ethanol or butanol, most preferably 100% methanol, to remove lower alcohol-soluble components therefrom, to obtain the intended lower alcohol-insoluble fraction.
  • a lower alcohol preferably methanol, ethanol or butanol, most preferably 100% methanol
  • the above-mentioned Hovenia dulcis Thunb is selected from the group comprising Hovenia dulcis Thunb. var. Koreana NAKAI, Hovenia dulcis Thunb. var tomentella Makino and Hovenia dulcis Thunb.
  • the lower alcohol-insoluble fraction thus obtained has the following characteristics: gel permeation chromatography (GPC) peaks at mean M.W. of 1,330,000 dalton (Da), 142,800 Da, 70,540 Da and 102,400 Da; IR (K r,.n ) absorption bands at 1000-1300 nm (ether, phenol, sulfbxide, vinyl peak); and a UV absorption at 200-300 nm (cyclic ring peak).
  • GPC gel permeation chromatography
  • the lower alcohol-insoluble fraction is dissolved in distilled water, an ion exchange column is charged with the resulting solution, polysaccharide fractions are eluted stepwise using solutions having increasing NaCl concentrations from 0 to 5M, dialyzed, concentrated, and lyophilized (see Fig. 1).
  • either a cation exchange resin or an anion exchange resin may be used.
  • exchange resins which can be used for this purpose are: strong acidic cation exchange resins such as AG 50W- x8, Amberlite IR-120 and Dowex 50W-x8; weak acidic cation exchange resins such as Amberlite IRC-50, Bio-Rex 70 and Duolite-436; weak basic cation exchange resins such as Amberlite IRA-67 and Dowex 3-x4A; strong basic cation exchange resins such as AG 2x8, Amberlite IRA-400 and Dowex 2-x8; modified cellulose cation exchange resins such as CM-Celluose and SE-Cellulose; a modified cellulose anion exchange resins such as DEAE Celluose; cationic sephadex-type resins such as G-25 and G-50 bead type cross-linked dextran resins; and modified bead-type ion exchange resins made from agarose such as Cepha
  • polysaccharide fractions are obtained according to the above isolation process and a polysaccharide that is eluted with 0.2 M NaCl solution shows the highest hepatoprotective activity.
  • polysaccharide obtained from Hovenia dulcis Thunb indigenous to China shows the following characteristics: It is composed of galactose, rhamanose, glucuronic acid, galacturonic acid, xylose, mannose and glucose in the ratio of 1 : 0.84: 1.05: 0.87: 0.89: 2.05.
  • the lower alcohol-insoluble fraction and the polysaccharide of the present invention may be employed as a health care food and a pharmaceutical agent for preventing or treating liver toxicity and liver diseases such as hepatitis, fatty liver and liver cirrhosis.
  • the present invention also provides a pharmaceutical composition for inhibiting alcohol dehydrogenase and lactic acid dehydrogenase, which comprises the lower alcohol-insoluble fraction of Hovenia dulcis Thunb or the polysaccharide isolated from Fraction 3 of the methanol-insoluble fractions as an active ingredient, in combination with pharmaceutically acceptable excipients, carriers or diluents.
  • the present invention provides a health care food comprising the extract and/or the polysaccharide described above.
  • the present invention also provides a pharmaceutical composition for preventing and treating liver diseases and hangover, which comprises the lower alcohol-insoluble fraction of Hovenia dulcis Thunb and the polysaccharide isolated therefrom as an active ingredient, in combination with pharmaceutically acceptable excipients, carriers or diluents. Additionally, the present invention provides a health care food comprising the extract and/or the polysaccharide described above.
  • the inventive pharmaceutical formulation may be prepared in accordance with any of the conventional procedures.
  • the active ingredient is preferably admixed or diluted with a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet or other container.
  • the carrier serves as a diluent, it may be a solid, semi-solid or liquid material acting as a vehicle, excipient or medium for the active ingredient.
  • the formulation may be in the form of a tablet, pill, powder, sachet, elixir, suspension, emulsion, solution, syrup, aerosol, soft and hard gelatin capsule, sterile injectable solution, sterile packaged powder and the like.
  • Suitable carriers, excipients, or diluents are lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoates, propylhydroxybenzoates, talc, magnesium stearate and mineral oil.
  • the formulation may additionally include fillers, anti-agglutinating agents, lubricating agents, wetting agents, flavoring agents, emulsifiers, preservatives and the like.
  • composition of the invention may be formulated so as to provide a quick, sustained or delayed release of the active ingredient after it is administrated to a patient, by employing any one of the procedures well known in the art.
  • the pharmaceutical formulation of the present invention can be administered via various routes including oral, transdermal, subcutaneous, intravenous and intramuscular introduction.
  • a typical daily dose of the above-mentioned fraction or polysaccharide isolated from Hovenia dulcis Thunb may range from about 0.01 to 10 g/kg body weight, preferably 1 to 5 g/kg body weight, and can be ao rrjriistered in a single dose or in divided doses.
  • the amount of the active ingredient actually administered ought to be determined in light of various relevant factors including the condition to be treated, the chosen route of administration, the age, sex and body weight of the individual patient, and the severity of the patient's symptom; and, therefore, the above dose should not be intended to limit the scope of the invention in any way.
  • the above-mentioned lower alcohol-insoluble fraction and the polysaccharide isolated therefrom can be added to food or beverage for preventing various liver diseases and hangover.
  • the amount of said fraction and/or polysaccharide that may be added to food or beverage for the purpose of preventing liver diseases or resolving hangover may generally range from about 0.1 to 15 w/w %, preferably 1 to 10 w/w % based on the total weight of food, and 1 to 30 g, preferably 3 to 10 g based on 100 ml of beverage.
  • the health care beverage composition of the present invention may contain other components, e.g., deodorants and natural carbohydrates as in conventional beverages.
  • natural carbohydrates are monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; conventional sugars such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol and erythritol.
  • a natural deodorant such as taumatin, levaudioside A, and glycyrrhizin, or a synthetic deodorant such as saccharin and aspartam may be used.
  • the amount of the above-described natural carbohydrate is generally in the range of about 1 to 20 g, preferably 5 to 12 g based on 100 ml of beverage.
  • compositions that may be added to the inventive food or beverage composition are various nutrients, vitamins, minerals, synthetic flavoring agents, coloring agents, pectic acid and its salt, alginic acid and its salt, organic acids, protective colloidal adhesives, pH controlling agents, stabilizers, preservatives, glycerin, alcohol, carbonizing agents, fruit juices and vegetable juices.
  • the molecular weight measurement was made using a GPC apparatus, equipped with a pump (spectra system, p2000 model), a guard column (TSK PWH, Tosoh Company), an Rl-detector (Shodex SE71 model), SEC (size exclusion chromatography) columns (TSK gel 3000pw, 4000pw, 5000pw (7.8x300 mi, Tosoh Company)), and a MALLS (multi angle laser light dispersion, Dawn DSP-F, Wyatt Technology Co.) detector, using a 0.02% sodium azide as developing solvent containing 0.15 M NaN0 3 at a flow rate of 0.5 m- ⁇ /min.
  • LDH lactic acid dehydrogenase
  • a 340-UV spectrometer Sigma Co., U.S.A.
  • Both the 3 H-Leucine(5 ⁇ Ci/plate) isotope used to determine the amount of synthesized protein having healing activity of the liver damaged by a hepatotoxic substance and the 3 H-Uridine isotope used to determine the amount of synthesized RNA were purchased from Sigma Co.
  • Absorbance variation measurement was conducted with a gas chromatography head-space analytical apparatus, HP 5890 gas chromatography (Hewlett Packard Company in USA) equipped with an FID (flame ionization detector), to determine the activity of alcohol dehydrogenase.
  • Example 1 Preparation a lower alcohol-insoluble fraction of Hovenia dulcis Thunb indigenous to Korea
  • Example 2 Analysis of the methanol-insoluble fraction of Hovenia dulcis Thunb
  • Example 1 The properties of the methanol-insoluble fractions obtained in Example 1 were analyzed as follows.
  • the melting temperature and the melting entalphy were determined by DSC (Differential Scanning Calorimeter, Seiko Instruments Inc. DSC 6100). Samples of the methanol-insoluble fraction of Hovenia dulcis Thunb indigenous to Korea was placed in an aluminum pan, sealed, and then heated from 20 ° C to 200 ° C at a rate of 10 U/rnin to obtain a melting heat absorption curve and the melting temperature, and the crystallinity of the sample was determined therefrom, respectively. The DSC scans of the methanol-insoluble fractions indigenous to Korea showed that a main peak starting at 164.9 °C and reaching a maximum melting temperature 185.3 °C. The methanol-insoluble fraction was formed to several carbohydrates (mp: 60 — 100 °C) and a minor amount of proteins (mp: 60- 100 ° C) (Fig. 2).
  • Acidic sugar moieties of the methylated product were reduced using LiB(C 2 H 5 ) 3 D (Super-Deupride, 1 m , Aldrich Company) in THF and the reduction product was recovered using a s 8x10 cartridge column(Sep-Pak). Subsequently, the treated sample was subjected sequentially to: hydrolysis at 121 ° C for 2 hours in 1.0 M TFA; reduction by NaBD 4 ; and acetylation. The resulting partially methylated alditol acetate was analyzed by GLC and GC-EIMS, and the peak areas were measured with an FID (flame ionization detector).
  • FID flame ionization detector
  • IR spectrum (Fig. 4) The sample was analyzed by IR spectroscopy (Vector 22 model, Bruker Analytician Messtechnik GMBH): resolution, 4.0; source, sphere; velocity, 6, 10 KHz; capture mode, dual wall/forward-backward condition).
  • IR (KBr, cm “1 ): ether, phenol, sulphoxide and vinyl peak (1000-1300nm; main 1039nm), aromatic ring peak (665, 939, 1313, 1663), hydroxyl peak (3435nm).
  • the sample was analyzed by UV-Vis spectroscopy (HP 8453 model, Hewlett Packard Company) and the result showed the maximum absorbance at 200-300 nm, suggesting the existence of a cyclic ring.
  • Example 3 Isolation of polysaccharides having hepatoprotective and hangover- resolving activities
  • Example 4 Characterization of the isolated fractions of Hovenia dulcis Thunb indigenous to Korea
  • Fractions 1 to 5 0.3 and 3 M NaCl were designated as Fractions 1 to 5, respectively.
  • the contents of total sugar and polyphenol components were determined by the phenol-sulfuric acid method (Dubois M. et al; Anal. Chem. 28, 350-356, 1956), and the result thus obtained is shown in Table 2.
  • Fraction 3 showed the highest hepatoprotective and hangover-resolving activities among the above fractions, and it was further characterized as below.
  • Example 5 Characterization of the isolated fractions of Hovenia dulcis Thunb indigenous to China
  • Example 2 The procedure of Example 1 was repeated to obtain a methanol-insoluble fraction (dry weight 61.5 g, yield: 4.1% w/w) and a methanol-soluble fraction (dry weight 133.5 g, yield: 8.9% w/w) from the fruit-peduncles of Hovenia dulcis Thunb indigenous to China.
  • the methanol-insoluble fraction was subjected to ion-exchange chromatography according to the method of Example 3 and the isolated fractions obtained by eluting with 0, 0.1, 0.2, 0.3 and 3 M NaCl were designated as Fractions 1 ' to 5 ', respectively.
  • Test Example 1 Hepatoprotective activity of the methanol-insoluble fraction of
  • Livers taken out from five-week old Sprague-Dawley rats were sliced into disc-shaped samples each having a diameter of about 0.8 mm and a thickness of 200 ⁇ m (wet weight: 18-22 mg), using a tissue cutter (Brendel/Vitron Co., USA).
  • the sliced samples were divided into 4 groups, two(group 3 and group 4) of which were treated with the methanol-insoluble fraction and the methanol-soluble fraction prepared in Example 1, respectively, each at a concentration of 200 /ig/m-E.
  • liver detoxification efficacy of the extract fraction of Hovenia dulcis Thunb was evaluated by determining the amount of synthesized protein according to the method described by Bonney et al. ("Some Characteristics and Function of Adult Rat Liver Primary Culture, in Gene Expression and Carcinogenesis in Cultured Liver", (1975) Gerschenson, E and Thompson, E. B. (Eds), Academic Press, New York, pp24-45). As shown in Figure 10a, it was demonstrated that the methanol-insoluble fraction has a much higher activity than the methanol-soluble fraction.
  • Fig. 10b was obtained by using the methanol- insoluble fraction or methanol-soluble fraction prepared in Example 5 according to the above method, wherein the methanol-insoluble fraction shows a much higher than the methanol-soluble fraction.
  • Test Example 1 The procedure of Test Example 1 was repeated using 500 ⁇ M of D- galactosamine and 1 ⁇ glml of LPS in place of carbon tetrachloride to measure the activity for restoring a damaged liver. As shown in Figure 11a, both of the methanol-insoluble and methanol-soluble fractions had significant restoration activities. Further, the result of Fig. 1 lb shows that the methanol-insoluble and methanol-soluble fractions obtained in Example 5 also exhibit significant restoration activities.
  • LDH lactic acid dehydrogenase
  • Test Example 2 Hepatoprotective activity of the polysaccharide isolated from the methanol-insoluble fraction of Hovenia dulcis Thunb
  • Livers taken out from five-week old Sprague-Dawley rats were sliced using a tissue cutter (Brendel/Vitron Co., USA) to obtain disc shaped samples each having a diameter of about 0.8 mm and a thickness of 200 ⁇ m (wet weight:
  • the sliced samples were divided into 7 groups, five of which were treated with the five polysaccharide fractions prepared in Example 4, respectively, each at a concentration of 200 ⁇ glm ⁇ (groups 1 to 5, respectively). Then, the sliced samples were surface-cultured for 1 hour under an atmosphere of
  • thermodynamic organ tissue cultivator (Sankyo Co., Japan).
  • bromobenzene was added to a concentration of 4 mM to each sample of groups 1 to 5 as well as to one of the remaining two groups (group 6).
  • the last remaining group (group 7) was treated with distilled water in place of bromobenzene (control).
  • liver detoxification efficacy ofeachpolysacchari.de fraction of Hovenia dulcis Thunb indigenous to Korea was evaluated by determining the amount of synthesized protein according to the method described by Bonney et al.
  • Fig. 15 showed that the alcohol concentration in blood samples taken out from the groups 1 to 3 are 0.058%, 0.032% and 0.028%, respectively. Namely, the methanol-insoluble fraction of Hovenia dulcis Thunb and the polysaccharide isolated therefrom lower the blood alcohol concentration by 44.8%) and 51.7%, respectively, as compared with the control group. This result demonstrates that the alcohol-insoluble fraction of Hovenia dulcis Thunb and the polysaccharide isolated therefrom have significant activities in lowering the alcohol concentration in blood.
  • Alcohol dehydrogenase activity in damaged liver was examined as follows.
  • the rats were suffocated by CO 2 and livers were taken out from the rats and washed with distilled water. Each liver sample was put in 10-fold volume of 0.1 M potassium phosphate buffer (pH 7.4) containing 0.154 M KC1 and homogenized with Teflon-glass homogenizer. The homogenized solution was subjected to centrifugation at 9,000 x g for 30 minutes at 4 ° C and the resulting supernatant was subjected to ultra-centrifugation at 110,000 x g for 1 hour at 4 ° C to obtain a supernatant cytosol fraction.
  • 0.1 M potassium phosphate buffer pH 7.4
  • Teflon-glass homogenizer Teflon-glass homogenizer
  • cytosole fraction 2-3 mg was added to a reaction mixture comprising 55 mM sodium pyrophosphate buffer(pH 7.4), 20 mM ethanol and 0.2 mM NAD.
  • concentration of NAD used in the reaction mixture was varied within the range of 0.025 mM to 2 mM.
  • the change of absorbance of the reaction mixture at 340 nm was measured for 3 minutes and the alcohol dehydrogenase activity was calculated from the slope of the absorbance curve.
  • the lower alcohol-insoluble fraction of the present invention and the polysaccharide isolated therefrom can be used in preparing a pharmaceutically effective powder, tablet, capsule, injection or liquid composition according to any one of the known conventional methods, as exemplified below.
  • Example 2 2 g of the dried extract obtained in Example 1 was mixed with 1 g of lactose to obtain a powder preparation, which was filled and sealed in a sealed package.
  • Example 1 100 mg of the dried extract obtained in Example 1, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were mixed and tabletted to obtain a tablet preparation.
  • Example 1 100 mg of the dried extract obtained in Example 1, 100 mg of the corn starch, 100 mg of lactose, 2 mg of magnesium stearate were mixed and filled in a gelatin capsule to obtain a capsule preparation.
  • the health care food was exemplarily prepared by the following method.
  • a scorched dried meal mixture of brown rice, barley, glutinous rice and Job's tear was pulverized and sieved to obtain grain particles of 60 mesh or less. Also, a mixture of black bean, black sesame and wild sesame was steamed, dried, scorched, pulverized and sieved to obtain seed particles of 60 mesh or less.
  • Example 1 The dried methanol-insoluble fraction of Hovenia dulcis Thunb obtained in Example 1 was pulverized and sieved to obtain particles of 60 mesh or less, which were mixed with the grain particles and seed particles in the following proportions to prepare a granule type health food.
  • Grains brown rice 30 w%, Job's tear 15 w%, barley 20 w%, Seeds : wild sesame 7 w%, black bean 8 w%, black sesame 7 w%, Dried powder of Hovenia dulcis Thunb. var. Koreana NAKAI : 3 w%,

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Abstract

L'invention concerne une composition pharmaceutique et un aliment diététique comprenant un extrait d'Hovenia dulcis Thunb insoluble dans l'alcool inférieur ou un polysaccharide isolé à partir de cet extrait, et présentant une puissante activité hépatoprotectrice et anti-gueule de bois.
PCT/KR2002/001449 2002-01-17 2002-07-31 Extrait d'hovenia dulcis thunb insoluble dans l'alcool inferieur et polysaccharide isole a partir de cet extrait WO2003059369A1 (fr)

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AU2002367003A AU2002367003A1 (en) 2002-01-17 2002-07-31 Lower alcohol-insoluble extract of hovenia dulcis thunb and a polysaccharide isolated therefrom

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KR2002/2772 2002-01-17
KR10-2002-0002772A KR100403721B1 (ko) 2001-01-31 2002-01-17 헛개나무로부터 분리된 간독성 및 숙취 해소 활성을 갖는저급알콜 불용성 추출 분획 및 다당체 물질 및 이를함유한 조성물

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Cited By (4)

* Cited by examiner, † Cited by third party
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CN101423558B (zh) * 2008-12-17 2011-08-10 四川大学 紫茎泽兰多糖及制备方法和用途

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WO2005072758A1 (fr) * 2004-01-31 2005-08-11 Kiyoung Kim Composition comprenant un extrait de plante hovenia dulcis, un extrait de lindera obtusiloba blume, ou un extrait de melange d'herbes de celles-ci
US7846484B2 (en) 2004-01-31 2010-12-07 Kiyoung Kim Composition comprising Hovenia dulcis thunb. extract, Lindera obtusiloba blume extract, or herbal mixture extract thereof
CN100396200C (zh) * 2005-11-24 2008-06-25 许滔 枳椇鲜果解酒饮料及其制备方法
WO2008002019A1 (fr) * 2006-06-28 2008-01-03 Il Hwan Cho Composition anti-gueule de bois
CN101423558B (zh) * 2008-12-17 2011-08-10 四川大学 紫茎泽兰多糖及制备方法和用途

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