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WO2007007993A1 - Composition pharmaceutique destinee a la prevention et au traitement de maladies du foie contenant un extrait de lonicera caerulea l. var. edulis - Google Patents

Composition pharmaceutique destinee a la prevention et au traitement de maladies du foie contenant un extrait de lonicera caerulea l. var. edulis Download PDF

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Publication number
WO2007007993A1
WO2007007993A1 PCT/KR2006/002668 KR2006002668W WO2007007993A1 WO 2007007993 A1 WO2007007993 A1 WO 2007007993A1 KR 2006002668 W KR2006002668 W KR 2006002668W WO 2007007993 A1 WO2007007993 A1 WO 2007007993A1
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Prior art keywords
extract
caerulea
liver
fruits
var
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PCT/KR2006/002668
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English (en)
Inventor
Ju Hwan Eum
Man Chul Suh
Dur Han Kwon
Byeong Ku Yoon
Wha Jeong Choi
Kyung Mi Kwon
Chan Soo Kim
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H & K Bioscience
Korea Polytechnic University
Korea Research Institute Of Bioscience And Biotechnology
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Application filed by H & K Bioscience, Korea Polytechnic University, Korea Research Institute Of Bioscience And Biotechnology filed Critical H & K Bioscience
Priority to JP2008521303A priority Critical patent/JP2009500446A/ja
Publication of WO2007007993A1 publication Critical patent/WO2007007993A1/fr

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    • 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)
    • A61K36/59—Menispermaceae (Moonseed family), e.g. hyperbaena or coralbead
    • 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)
    • A61K36/35—Caprifoliaceae (Honeysuckle family)
    • A61K36/355—Lonicera (honeysuckle)
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/08—Antiepileptics; Anticonvulsants

Definitions

  • the present invention relates to a pharmaceutical composition having preventative and therapeutic effects on liver diseases, comprising an extract from Lonicera caerulea L. var. edulis.
  • the liver situated between the digestive system and the systemic circulatory system, plays important roles in protecting the whole body from foreign toxic substances and in the metabolism of exogenous materials. Since the exogenous materials taken up by the body initially enter the liver to be filtered, the liver has a high risk of being exposed to numerous toxic substances as well as nutrients. Thus, the liver is highly vulnerable to damage relative to other organs.
  • Liver diseases are classified into two major types according to cause: one is toxic liver disease caused by the excessive ingestion of alcohol or the like, and the other is viral liver disease caused by viral infection.
  • Viral liver diseases arise from infection with hepatitis B virus, hepatitis C virus, or the like. Recently, toxic liver disease is increasing due to food, medicaments, medicinal herbal substances, alcohol, and the like. Liver diseases are difficult to diagnose in early stages due to the absence of subjective symptoms. By the time individuals develop subjective symptoms, the liver has suffered great damage. The liver is an organ which has greater ability to recover its full function than other organs. However, it is difficult to restore normal liver function when hepatocytes have already been transformed.
  • silymarin Since silymarin has been domestically introduced as a therapeutic agent for liver damage in the 1970' s, many pharmaceutical preparations containing it as a major ingredient have been developed and are now available on the market. Pharmaceutical preparations containing silymarin as a major ingredient have already been widely applied for the clinical purpose of treating liver diseases. However, the major ingredient silymarin has a drawback in that it is not highly water-soluble, and thus has a low uptake in the body when orally administered. At present, silymarin preparations have been used merely in auxiliary therapy for liver diseases, such as toxic liver diseases, chronic hepatitis and liver cirrhosis. Thus, there is a need for the development of drugs capable of rapidly restoring the normal function of hepatocytes .
  • Korean Pat. Registration No. 80759 discloses fermented milk, which is useful for maintaining and improving liver function, and a method of preparing the same.
  • Korean Pat. Laid-open Publication No. 2003-0027615 discloses a functional food composition containing an extract from fruits of Hovenia dulcis Thunb, the composition having effects of enhancing liver function and relieving hangover symptoms.
  • Korean Pat. Laid-open Publication No. 2003-0011818 discloses the use of an extract from Eleutherococcus senticosus in the production of functional rice coated therewith.
  • 2003-0063308 discloses a therapeutic agent for hepatitis B comprising an extract from the medicinal herb Phyllanthus urinaria, and a method of preparing the same.
  • Korean Pat. Laid-open Publication No. 2004-0018733 discloses a composition for treating viral liver diseases comprising an extract from Ixeris sonchifolia.
  • the present inventors conducted intensive and thorough research to obtain from natural materials a substance having good therapeutic activity against liver diseases, other than conventional therapeutic compositions as described above.
  • the research resulted in the finding that when a damaged hepatic cell line was dosed with an extract from Lonicera caerulea L. var. edulis, cell growth was stimulated, bringing about the restoration of damaged liver function, which was determined by remarkable decreases in GOT and GPT levels as biochemical markers for liver function.
  • a composition comprising the extract from Lonicera caerulea L. var. edulis has good preventive and therapeutic effects on liver diseases, such as hepatitis, liver cirrhosis and fatty liver, thereby leading to the present invention.
  • the present invention aims to provide a pharmaceutical composition having preventative and therapeutic effects on liver diseases, comprising an extract from Lonicera caerulea L. var. edulis.
  • FIG. 1 shows the effects of an extract from fruits of Lonicera caerulea L. var. edulis on the growth of HepG2 cells .
  • Fig. 2 shows the effects of ethyl alcohol on the growth of HepG2 cells.
  • Fig. 3 shows the comparison of absorbance of a well treated with ethyl alcohol alone and wells treated with ethyl alcohol and then an extract from fruits of Lonicera caerulea L. var. edulis with that of a control well not treated with either ethyl alcohol or the extract from fruits of Lonicera caerulea L. var. edulis.
  • Fig. 4 shows the restoration of liver function by administration of an extract from fruits of Lonicera caerulea L. var. edulis, which was observed on a SPOTCHEMTM II strip.
  • Fig. 5 shows the results of histochemical analysis on the effects of an extract from fruits of Lonicera caerulea L. var. edulis and silymarin on acute hepatitis when they were administered into a transgenic mouse model of acute hepatitis. [Best Mode]
  • the present invention relates to a pharmaceutical composition having preventative and therapeutic effects on liver diseases, comprising an extract from Lonicera caerulea L. var. edulis.
  • extract refers to an active ingredient isolated from a natural material.
  • the extract may be obtained by an extraction process using water, an organic solvent, or a solvent mixture thereof, and includes dry powder of the extract or all forms formulated therefrom.
  • Lonicera caerulea L. var. edulis refers to all organs, for example, roots, branches, stems, leaves, flowers and fruits, of natural, hybrid or variant types of Lonicera caerulea L. var. edulis, but preferably indicates fruits of Lonicera caerulea L. var. edulis.
  • Lonicera caerulea L. var. edulis is a dicotyledonous plant belonging to the Family Caprifoliaceae of the Order Rubiales. It is a deciduous shrub that grows to 1.5 m tall, is densely branched, and has shield-shaped bracts at nodes of twigs.
  • the inner part of branches is white.
  • the leaves are opposite, lanciform or elliptic and blunt- or sharp-ended, lack teeth on the margins, have short hairs on the margins and surface, and have many wooly hairs underneath.
  • the flowers usually have short stalks, which arise from leaf axils, have trumpet-shaped creamy white corollas, and bloom in summer.
  • Each calyx contains five toothed sepals.
  • the corollas are yellowish white, cylindrical campanulate, 1.2-1.5 cm long, and slightly hairy.
  • the stamens are shorter than styles and have no hairs, and the two ovaries are fused together.
  • the fruits are oval or nearly circular, ripen to purplish black between July and October, and are covered with white powder.
  • This deciduous shrub is an arctic plant that is widespread in Siberia, Sakhalin, the Northern region of China, Cambodia, North Korea, and the like.
  • Lonicera caerulea L. var. edulis was not investigated prior to the present invention for effects of stimulating the growth of hepatocytes and improving liver function due to the stimulatory effect.
  • liver function can be objectively evaluated by measuring the degree of stimulation of growth rates of damaged hepatocytes due to composition administration and levels of hepatic enzymes, aspartate aminotransferase (AST, also known as GOT) and alanine aminotransferase (ALT, also known as GPT) .
  • AST aspartate aminotransferase
  • ALT alanine aminotransferase
  • GOT composition administration and levels
  • ALT and AST are enzymes present in hepatocytes. When hepatocytes are damaged or disrupted, these enzymes are released therefrom, leading to an increase in concentrations thereof in the blood.
  • ALT and AST levels are used as biochemical indicators for liver diseases caused by liver cell damage.
  • a human liver cell line, HepG2 was damaged with ethyl alcohol and then dosed with an extract from Lonicera caerulea L. var. edulis, and stimulated growth rates were compared with those of a control not dosed with the extract from Lonicera caerulea L. var. edulis (also referred herein to simply as "L. caerulea extract”) .
  • the L. caerulea extract was found to stimulate the growth of damaged hepatocytes in a dose-dependent manner.
  • the L. caerulea extract stimulated the cell growth by about 27% at 0.25 mg/ml and about 54% at 0.5 mg/ml.
  • ALT and AST levels were measured to determine whether liver function was enhanced.
  • the administration of L. caerulea extract resulted in a reduction of ALT and AST levels in HepG2 cells.
  • L. caerulea extract was administered into a transgenic mouse of acute hepatitis, it exhibited greater ability to restore liver function by 25% more than the conventional drug silymarin.
  • liver diseases refers to all diseases that bring about decreased liver function. Liver diseases are caused by viruses (e.g., hepatitis virus A, B, C, D or E) , alcohol, drugs (antituberculosis drugs, aspirin, antibiotics, anesthetics, antihypertensive drugs, oral contraceptives, etc.), congenital metabolic disorders, and the like. Detailed examples of liver diseases are liver hepatitis, liver cirrhosis and fatty liver. Liver hepatitis includes chronic and acute liver hepatitis.
  • prevention refers to all actions that inhibit or delay the reduction of liver function through composition administration.
  • treatment refers to all actions that restore or beneficially change liver function and liver regeneration through composition administration.
  • the administration of the present composition may prevent and treat liver hepatitis, liver cirrhosis, fatty liver, and other liver diseases, as well as symptoms or complications caused by the diseases .
  • symptoms of liver diseases which can be prevented and treated by the L. caerulea extract, include fatigue, vomiting, decreased appetite, abdominal pain, and jaundice.
  • complications include edema, ascites, gastrointestinal bleeding, esophageal variceal bleeding, and hepatic encephalopathy (hepatic coma) .
  • the L. caerulea extract is prepared using extraction with water, an organic solvent, or a solvent mixture thereof.
  • the resulting extract may be used as it is or after being concentrated and/or dried.
  • an organic solvent such as methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethylacetate, butylacetate, dichloromethane, N, N- dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3- butylene glycol, propylene glycol, or a solvent mixture thereof. Since the degree of extraction and loss of an effective ingredient may vary depending on the organic solvent used, a suitable organic solvent must be selected and employed.
  • the extraction method is not specifically limited, and includes cold precipitation, ultrasonic extraction, and reflux extraction.
  • the solvent extraction may further include a step of filtering the extract to remove suspending solid particles.
  • the removal of particles may be achieved using cotton, nylon, and the like, or using ultrafiltration, freezing filtration, centrifugation, and the like, but the present invention is not limited to the examples .
  • the concentration of the extract may be performed using reduced pressure, reverse osmosis, and the like.
  • the concentrate is dried by freeze drying, vacuum drying, hot wind drying, spray drying, drying under reduced pressure, foam drying, high frequency drying, infrared drying, and the like, but the present invention is not limited to the examples.
  • the present method may further include a step of pulverizing the final dried extract.
  • the extract may be optionally subjected to a fractionation process.
  • the extract is suspended in distilled water, and extracted using a nonpolar organic solvent, such as hexane, ether, dichloromethane, chloroform, ethylacetate, or a solvent mixture thereof to separate a nonpolar solvent-soluble layer.
  • a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethylacetate, or a solvent mixture thereof to separate a nonpolar solvent-soluble layer.
  • the obtained nonpolar solvent-soluble layer is concentrated and/or dried.
  • the L is a detailed practice, the L.
  • caerulea extract of the present invention was obtained by hot water extraction, cold water extraction, ultrasonic extraction or reflux extraction, preferably reflux extraction, using water, Ci to C 4 lower alcohol or a solvent mixture thereof weighing 5 to 25 times, preferably 7 to 15 times as much as the dry- weight (kg) of Lonicera caerulea L. var. edulis, preferably fruits thereof.
  • the extraction was carried out at 20 ° C to 100 ° C, preferably 60 ° C to 100 ° C, for a period ranging from 0.5 hrs to 2 days, preferably 1 hr to 1 day, and was serially performed 1-5 times, preferably 2-3 times.
  • the extract was passed through filter paper.
  • the filtrate was concentrated under reduced pressure using a rotary vacuum concentrator at 20 ° C to 100 ° C, preferably 50 ° C to 70 ° C, and dried, thereby yielding the L. caerulea extract in powder form according to the present invention.
  • the L. caerulea extract in the powder form may be used as it is or after being dissolved in a solvent at a predetermined concentration .
  • the L. caerulea extract is safe and does not cause side effects or stimulate resistance thereto because it contains substances obtained from natural material as effective ingredients.
  • the L. caerulea extract has an advantage in that it is able to be administered for a long period of time. Actually, an acute toxicity test in mice revealed that the L. caerulea extract is not toxic.
  • the composition may be applied to humans, as well as livestock including cattle, horses, sheep, pigs, goats, antelopes and dogs .
  • the L. caerulea extract is contained in the pharmaceutical composition for preventing and treating liver diseases in an amount of 0.01% to 100%, and more preferably 1% to 80% by weight based on the total weight of the composition.
  • the composition may further include an additive which does not increase efficacy but is commonly used in pharmaceutical compositions to enhance flavor, taste, color, or the like.
  • the composition may further include inorganic and organic additives, such as vitamins Bi, B 2 , B 6 , C and E, niacin, carnitine, betaine, folic acid, pantothenic acid, biotin, zinc, iron, calcium, chrome, magnesium, and mixtures thereof.
  • the composition may be used alone, or may further include a conventionally used substance having therapeutic activity against liver diseases .
  • the composition may include a pharmaceutically acceptable carrier, and may be formulated into dosage forms for human or veterinary use. According to the intended use, the composition may be formulated into a variety of ordinary forms suitable for oral, parenteral and topical administration. Oral solid preparations, such as powders, granules, tablets and capsules, may be prepared using binders, lubricants, integrators, excipients, solubilizers, dispersing agents, stabilizers, suspending agents, pigments, flavors, and the like.
  • Oral liquid preparations such as suspensions, solutions, emulsions and syrups, may be prepared with commonly used simple diluents, water and liquid paraffin, as well as humectants, sweeteners, aromatics, preservatives, and the like.
  • injectable preparations may be prepared by mixing buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, and the like.
  • Such a composition may be presented in unit-dose (single dose) or multiple dose (several doses) containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example water for injections, immediately prior to use.
  • a sterile liquid carrier for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
  • the present invention relates to a method of preventing and treating liver diseases, comprising administering a composition comprising the L. caerulea extract to a patient.
  • patient refers to a human or an animal, such as horses, sheep, pigs, goats, camels, antelopes and dogs, which has a disease characterized by reduced or damaged liver function, for example, hepatitis, liver cirrhosis, or fatty liver, and symptoms of the disease may be relieved through the administration of the present composition.
  • the aforementioned diseases may be effectively prevented and treated by administering the composition comprising the L. caerulea extract of the present invention to a patient.
  • the present composition may be administered in combination with a conventional therapeutic agent for liver diseases.
  • administration refers to the introduction of a predetermined material into a patient using any suitable method.
  • the present composition may be orally or parenterally administered via any of the common routes, as long as it is able to reach the desired tissue.
  • the composition may be administered using a certain apparatus capable of transporting active substances into target cells.
  • the present composition may be administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount refers to an amount sufficient for treating or preventing diseases, which is commensurate with a reasonable benefit/risk ratio applicable for medical treatment or prevention.
  • An effective dosage amount of the composition may be determined depending on the patient's gender and age, the severity of the illness, drug activity, sensitivity to drugs, administration time, administration routes and excretion rates, treatment duration, drugs used in combination with the composition; and other factors known in medicine.
  • the present composition may be administered as a sole therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. This administration may be provided in single or multiple doses.
  • the extract prepared according to the preparation method of the present invention is preferably administered orally or intravenously.
  • a single dose of the extract is preferably 1 to 20 mg/kg adult for oral administration, and preferably 1 to 20 mg/kg adult for intravenous administration.
  • the dosage for a specific patient may vary depending on the patient's gender, age, health state and diet, administration time, administration modes, co- administered drugs, and severity of illness.
  • EXAMPLE 1 Preparation of an extract from fruits of Lonicera caerulea L. var. edulis
  • EXAMPLE 2 Evaluation of the effect of the extract from fruits of Lonicera caerulea L. var. edulis on the growth of hepatocytes
  • HepG2 cells were seeded in a 96-well plate at a density of IXlO 4 cells per well, and incubated in a culture medium supplemented with 10% fetal bovine serum (FBS) for 16 hrs. After the 96-well plate was washed with physiological saline, the extract from L. caerulea fruits, which was diluted in culture medium in amounts of 0, 0.125, 0.25, 0.5 and 1 mg/ml, was added to each well. After incubation for 48 hrs, the cell number in each well was measured using a SRB method.
  • FBS fetal bovine serum
  • the culture medium containing the extract from L. caerulea fruits was removed from the 96-well plate.
  • the 96-well plate was washed with physiological saline, and the cells in each well were fixed with 70% acetone for 20 min.
  • the fixed cells were dried, stained with a SRB solution for 30 min, and washed with a 1% acetic acid solution five times to eliminate unbound SRB.
  • 10 mM Tris was added to the cells to dissolve cellular proteins and unbound SRB, and absorbance was measured at 562 nm using a spectrophotometer. The results were expressed as a percentage by comparing the absorbance of each well treated with the extract from L.
  • caerulea fruits with that of a control well not treated with the extract from L. caerulea fruits (Fig. 1) .
  • Fig. 1 When cells were dosed with the extract from L. caerulea fruits at up to 0.5 mg/ml, their number stayed the same or increased slightly, or their growth was little affected in comparison with the well not treated with the extract from L. caerulea fruits.
  • EXAMPLE 3 Evaluation of the effect of ethyl alcohol on the growth of hepatocytes
  • HepG2 cells were seeded in a 96-well plate at a density of IXlO 4 cells per well and incubated in a culture medium supplemented with 10% FBS for 16 hrs . After the 96- well plate was washed with physiological saline, ethyl alcohol, which was diluted in a culture medium in amounts of 0, 0.5, 1.0, 1.5 and 2.0% (v/v) , was added to each well in which HepG2 cells were cultured. After incubation for 48 hrs, the cell number in each well was measured using a SRB method.
  • the culture medium was removed, and the 96-well plate was washed with physiological saline.
  • the cells in each well were then fixed with 70% acetone for 20 min.
  • the fixed cells were dried, stained with an SRB solution for 30 min, and washed with a 1% acetic acid solution five times to eliminate unbound SRB.
  • 10 mM Tris was added to the cells to dissolve cellular proteins and unbound SRB, and absorbance was measured at 562 nm using a spectrophotometer. The results were expressed as a percentage by comparing the absorbance of each well treated with ethyl alcohol with that of a control well not treated with ethyl alcohol (Fig. 2) .
  • Ethyl alcohol was found to stimulate cell growth at concentrations of up to 1%, but reduced cell number at concentrations of 1.5% or higher.
  • EXAMPLE 4 Evaluation of the ability of the extract from L. caerulea fruits to restore the growth of cells whose growth is suppressed by ethyl alcohol
  • HepG2 cells were seeded in a 96-well plate at a density of IXlO 4 cells per well, and incubated in a culture medium supplemented with 10% FBS for 16 hrs . After the 96- well plate was washed with physiological saline, ethyl alcohol, which was diluted in culture medium in 1.5% (v/v) , was added to each well in which HepG2 cells were cultured. Cell damage was induced using ethyl alcohol for 24 hrs. Thereafter, the culture medium containing ethyl alcohol was removed, and the extract from L. caerulea fruits, which was diluted in culture medium in amounts of 0, 0.125, 0.25, 0.5 and 1 mg/ml, was added to each well.
  • Restoration rate of cell growth (group dosed with the extract from L. caerulea fruits - group dosed with ethyl alcohol alone) / (non treatment group - group dosed with ethyl alcohol alone)
  • Hep3B cells were seeded in a 96-well plate at a density of 2X10 4 cells per well, and were incubated for 12 hrs to allow them to adhere to the bottom of the plate. After the culture supernatant was removed, the plate was washed with physiological saline. 5% Ethyl alcohol in culture medium was added to each well, and incubated for 12 hrs to induced cell damage. After the culture supernatant was removed from each well, a culture medium containing the extract from L. caerulea fruits (0.3 mg/ml) and a culture medium not containing the extract were individually added to the well in which cell damage was induced by ethyl alcohol.
  • ALT and AST levels were measured in wells treated with ethyl alcohol or not using a SPOTCHEMTM II strip from the Array Company in Japan.
  • the SPOTCHEMTM II strip enables the measurement on a single strip of all of amounts of ALT (GOT) , AST (GPT) , blood urea nitrogen, glucose, total cholesterol and total bilirubin.
  • the region measuring ALT and AST levels is present as a yellowish white zone in the strip. Increased ALT and AST levels in a sample change the yellowish white zone to dark blue.
  • the culture supernatant of each well was collected and loaded onto the SPOTCHEMTM II strip, and the strip was observed for color change in the yellowish white zone (Fig. 4) .
  • the control was a culture supernatant of Hep3B cells not treated with ethyl alcohol or with the extract from L. caerulea fruits.
  • the yellowish white zone was changed to light blue due to small amounts of ALT and AST present in the serum of culture medium.
  • a culture supernatant of a well treated with 5% ethyl alcohol was applied to the strip, the yellowish white zone was changed to dark blue, indicating rapidly increased AST and ALT activity.
  • cells were dosed with ethyl alcohol to induce cell damage and then with the extract from L. caerulea fruits (0.3 mg/ml) , the yellowish white zone changed to light blue.
  • the hot water extract or hot water alcohol extract prepared in Example 1 was dissolved in distilled water, and administered to mice (ten per group) at a dosage of 500 mg/kg. Then, the mice were monitored for 7 days. No death was observed, indicating that the extract was not toxic.
  • EXAMPLE 7 Evaluation of the effect of the extract from L. caerulea fruits on acute hepatitis induced in mice
  • mice In order to determine whether the extract from L. caerulea fruits has the ability to restore liver function in acute hepatitis-induced mice, this test was carried out as follows. Forty mice (ICR) were divided into four groups
  • Group A was not dosed with any drug.
  • Group B was allowed to ingest olive oil alone.
  • Group C was orally dosed with 100 ⁇ JL of silymarin (Sigma) , which was dissolved in olive oil at 20 mg/ml, for 3 days.
  • Group D was orally dosed with 100 ⁇ i of the extract from L. caerulea fruits prepared in Example 1, which was dissolved in distilled water at 500 mg/ml, for 3 days.
  • Groups A
  • mice B, C and D all received only water. Then, 100 ⁇ Jt of 1% carbon tetrachloride in olive oil was intraperitoneally injected into mice of Groups B, C and D. After 18 hrs, blood samples were collected from the mice in Groups A, B, C and D. AST and ALT activities were determined in the mouse blood according to the same method as in Example 5, and mean AST and ALT activities were compared among groups . In Group A, which was not treated with carbon tetrachloride, mean levels of AST and ALT activities were 23 IU/L and 37 IU/L, respectively.
  • caerulea fruits decreased by 230 IU/L and 207 IU/L, respectively, in comparison with those in mice pretreated with silymarin. With respect to AST and ALT levels, silymarin restored liver function by 63.8%, and the extract from L. caerulea fruits by 89.0%. These results indicate that the extract from L. caerulea fruits had ability to restore damaged liver function which was 25% better than that of silymarin.
  • EXAMPLE 8 Comparison of the results of histochemical analysis between the extract from L. caerulea fruits and silymarin in acute hepatitis-induced mice
  • Example 7 Histochemical analysis was performed with mouse liver tissues from Test Groups A, B, C and D in Example 7, whose mean AST and ALT levels were already determined in Example 7.
  • the liver tissue from each mouse was sectioned, fixed in 10% formalin, immersed in different concentrations of ethyl alcohol to be dehydrated, and finally embedded in paraffin.
  • the paraffin-embedded tissue was sectioned to a size of 4-5 ⁇ m. Each paraffin section was covered with a slide glass, stained with hematoxylin and eosin, and observed under an optical microscope (Fig. 5) . Mice treated with carbon tetrachloride showed multiple scattered necrotic areas in the liver tissue, which were not stained with hematoxylin and eosin (Fig.
  • liver tissues from mice pretreated with silymarin and mice pretreated with the extract from L. caerulea fruits (Fig. 5, B and C panels) .
  • necrotic reaction rarely occurred compared to normal liver tissue (Fig. 5, D panel) .
  • Fig. 5, D panel a panel of liver tissue from mice pretreated with the extract from L. caerulea fruits
  • hepatitis symptoms Three volunteers having hepatitis symptoms were orally dosed with the extract from L. caerulea fruits (1 g) , prepared in Example 1, two times everyday for a period of 10 days or 20 days. Then, levels of the liver enzymes ALT and AST were measured. After the period of 10 days or 20 days, all of the three subjects showed a decrease in ALT and AST levels.
  • Subject 1 male age forty
  • AST and ALT levels decreased from 32 to 11.1 and from 73 to 14.3, respectively; 20 days after administration, AST and ALT levels decreased to 10.3 and 10.6, respectively.
  • Subject 2 male age thirty five
  • AST and ALT levels decreased from 53 to 2.25 and from 96 to 35, respectively; 60 days after administration, AST and ALT levels decreased to 2.25 and 5.75, respectively.
  • Subject 3 female age thirty five
  • AST and ALT levels decreased from 69 to 26 and from 72 to 29, respectively.
  • Preparation Example 1 Preparation of powder capsules The 100 mg of the hot water extract prepared in Example 1 were mixed with 14.8 mg of lactose, 3 mg of crystalline cellulose, and 0.2 mg of magnesium stearate. The mixture was filled into a No. 5 capsule using a suitable device. The components of powder capsules were summarized below.
  • Effective ingredient 100 mg Lactose: 14.8 mg Crystalline cellulose: 3 mg Magnesium stearate: 0.2 mg
  • An injectable solution containing 100 mg of the hot water extract prepared in Example 1 was prepared as follows . 100 mg of the hot water extract prepared in Example 1, 600 mg of sodium chloride and 100 mg of ascorbic acid were dissolved in distilled water, and the final volume was adjusted to 100 ml. The resulting solution was placed in a bottle and sterilized at 120 ° C for 30 min.
  • the components of the injectable solution were as follows .
  • Effective ingredient 1000 mg Sodium chloride: 6000 mg
  • Powders were prepared with the following composition according to the powder preparation method of the Korean Pharmacopeia .
  • Effective ingredient 100 mg Lactose: 100 mg Talc: 10 mg
  • Water-soluble fraction of effective ingredient 100 mg Lactose: 100 mg Talc: 10 mg
  • Tablets were prepared with the following composition according to the tablet preparation method of the Korean Pharmacopeia . 1) Each tablet contained
  • Effective ingredient 100 mg Corn starch: 100 mg Lactose: 100 mg Magnesium stearate: 2 mg
  • Water-soluble fraction of effective ingredient 100 mg Corn starch: 100 mg Lactose: 100 mg Magnesium stearate: 2 mg
  • Capsules were prepared with the following composition according to the capsule preparation method of the Korean Pharmacopeia . 1) Each capsule contained
  • Injections were prepared with the following composition according to the injectable preparation method of the Korean Pharmacopeia.
  • Water-soluble fraction of effective ingredient 50 mg Sterile distilled water for injection: suitable amount pH controller: suitable amount
  • Preparation Example 7 Preparation of solutions Solutions were prepared with the following composition according to the solution preparation method of the Korean Pharmacopeia .
  • Water-soluble fraction of effective ingredient 100 mg Isomerized sugar: 10 g Mannitol: 5 g Purified water: suitable amount
  • the composition containing the L. caerulea extract according to the present invention has good ability to restore liver function and liver regeneration with no side effects such as toxicity.
  • the present composition can be effectively used as a preventive and therapeutic agent for liver diseases .

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Abstract

L'invention concerne une composition pharmaceutique présentant des effets préventifs et thérapeutiques sur les maladies du foie, notamment un extrait de Lonicera caerulea L. var. edulis . La composition présente des effets préventifs et thérapeutiques sur l'hépatite, la cyrrhose du foie, la stéatose hépatique, et analogue.
PCT/KR2006/002668 2005-07-13 2006-07-07 Composition pharmaceutique destinee a la prevention et au traitement de maladies du foie contenant un extrait de lonicera caerulea l. var. edulis WO2007007993A1 (fr)

Priority Applications (1)

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JP2008521303A JP2009500446A (ja) 2005-07-13 2006-07-07 ケヨノミの抽出物を含む肝臓疾患予防及び治療効果を有する薬剤学的組成物

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KR1020050063026A KR100699790B1 (ko) 2005-07-13 2005-07-13 댕댕이나무 추출물을 포함하는 간 질환 예방 및 치료효과를 가지는 약제학적 조성물
KR10-2005-0063026 2005-07-13

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WO2007007993A1 true WO2007007993A1 (fr) 2007-01-18

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JP (1) JP2009500446A (fr)
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Cited By (2)

* Cited by examiner, † Cited by third party
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CN105636603A (zh) * 2013-09-24 2016-06-01 H&K生物科学株式会社 包含蓝靛果果实的提取物作为活性成分的预防或治疗甲状腺疾病的药物组合物
CN107582787A (zh) * 2017-09-25 2018-01-16 黄经卫 一种治疗肝硬化的中草药

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WO2015046743A1 (fr) 2013-09-24 2015-04-02 주식회사 에이치앤케이바이오사이언스 Composition pharmaceutique pour prévenir ou traiter des maladies thyroïdiennes, contenant de l'extrait de fruit de lonicera caerulea l. var. edulis comme principe actif
WO2016108400A2 (fr) * 2014-12-31 2016-07-07 주식회사 에이치앤케이바이오사이언스 Composition pour la peau, contenant des extraits de fruits de lonicera caerulea en tant que principe actif
KR102038887B1 (ko) * 2017-11-30 2019-10-31 주식회사 아리바이오 댕댕이나무열매 추출물을 포함하는 비만, 고지혈증 또는 지방간의 예방 또는 치료용 약제학적 조성물 및 이의 제조방법
KR20220117436A (ko) 2021-02-17 2022-08-24 (주)사랑과 선행 댕댕이나무 열매 추출물을 포함하는 근감소성 비만 개선용 조성물
KR20220117437A (ko) 2021-02-17 2022-08-24 가천대학교 산학협력단 댕댕이나무 열매 추출물을 포함하는 장내 균총 개선용 조성물
CN114605852B (zh) * 2022-03-30 2023-04-14 吉林大学 一种双重低温等离子体辅助酶解蓝靛果提取花色苷的方法

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CN105636603A (zh) * 2013-09-24 2016-06-01 H&K生物科学株式会社 包含蓝靛果果实的提取物作为活性成分的预防或治疗甲状腺疾病的药物组合物
CN107582787A (zh) * 2017-09-25 2018-01-16 黄经卫 一种治疗肝硬化的中草药

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