WO2023048504A1 - Composition pour prévenir ou traiter des maladies associées à une faiblesse musculaire ou une sarcopénie, contenant un extrait de rosa davurica pallas en tant que principe actif - Google Patents

Composition pour prévenir ou traiter des maladies associées à une faiblesse musculaire ou une sarcopénie, contenant un extrait de rosa davurica pallas en tant que principe actif Download PDF

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WO2023048504A1
WO2023048504A1 PCT/KR2022/014275 KR2022014275W WO2023048504A1 WO 2023048504 A1 WO2023048504 A1 WO 2023048504A1 KR 2022014275 W KR2022014275 W KR 2022014275W WO 2023048504 A1 WO2023048504 A1 WO 2023048504A1
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extract
sarcopenia
muscle weakness
preventing
composition
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PCT/KR2022/014275
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English (en)
Korean (ko)
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연성흠
김현준
박현진
필감방
문효원
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(주)휴온스
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
    • 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)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/738—Rosa (rose)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system

Definitions

  • the present invention relates to a composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia, comprising an extract of wild genus japonica as an active ingredient.
  • a representative physiological change according to aging is a decrease in muscle mass and strength. It is known that muscle mass loss occurs after the age of 40 and decreases by about 8% per 10 years until the age of 70.
  • Muscle weakness disease or sarcopenia causes direct muscle strength reduction, which not only increases the risk of death due to a decrease in various physical functions and disabilities, but also reduces metabolism and lowers immunity, such as high blood pressure and diabetes. It is also a factor that increases the prevalence of metabolic diseases such as arthritis, obesity, and cancer.
  • Myostatin is a protein that regulates the growth of skeletal muscle and belongs to the transforming growth factor- ⁇ (TGF- ⁇ ) family and is also known as growth and differentiation factor-8 (GDF-8). It is known that controlling the function of the myostatin gene in mice through gene knock-out technology shows 2 to 3 times more skeletal muscle phenotypes than normal mice.
  • Methods for inhibiting the function of myostatin at the molecular level include MSTN prodomain, activin type II receptor, follistatin, and follistatin-related proteins.
  • Myostatin which inhibits the growth of skeletal muscle, has been studied in relation to this decrease in muscle mass. Antibodies were prepared to inhibit only the function of myostatin, but side effects appeared, and anti-myostatin materials using natural products Research on this is in progress, but most of the studies are still at the cellular level, and there are no studies that have reached clinical trials.
  • the present invention is to provide a pharmaceutical composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia, which contains an extract of wild chives, a fraction thereof, or a fermented product thereof as an active ingredient.
  • the present invention provides a pharmaceutical composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia, containing an extract, a fraction thereof, or a fermented product thereof as an active ingredient.
  • the raw fever tree extract can be extracted from any one or more parts selected from the group consisting of leaves, stems, flowers, fruits, and outposts of the tree.
  • the raw heat tree extract can be extracted using water, an organic solvent, or a mixture thereof as a solvent for the raw heat tree tree.
  • the raw wild cypress extract, a fraction thereof, or a fermented product thereof may inhibit myostatin activity.
  • Concentration of the extract of the wild chives tree in the composition may be 0.1 ⁇ g/mL to 100 ⁇ g/mL.
  • the disease related to muscle weakness may be one or more selected from the group consisting of muscular dystrophy, myasthenia gravis, muscular dystrophy, myasthenia, hypotonia, muscle weakness, muscle degeneration, muscular dystrophy, amyotrophic lateral sclerosis, myasthenia gravis, hypotonia and cachexia. .
  • a health functional food composition for strengthening muscle strength or preventing or improving sarcopenia which contains an extract of genus japonica, a fraction thereof, or a fermented product thereof as an active ingredient.
  • a food composition for strengthening muscle strength or preventing or improving sarcopenia which contains an extract of wild chives, a fraction thereof, or a fermented product thereof as an active ingredient.
  • the present invention relates to a composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia, containing as an active ingredient a natural ingredient extract, a fraction thereof, or a fermented product thereof. Since it can inhibit ausstatin activity, it can be effectively used for strengthening muscle strength or preventing or treating sarcopenia.
  • Figure 1 is a graph analyzing the cytotoxicity of CAGA 12 -Luc-expressing 293T cell line according to the treatment of various concentrations of wild chives (whole plant) extract (12.5, 25, 50, 100 ⁇ g/mL).
  • Figure 2 (a) shows that CAGA 12 -Luc-expressing 293T cell line was treated with recombinant myostatin and various concentrations of wild chives extract (0.2, 0.4, 0.8, 1.6, 3.1, 6.3, 12.5, 25 ⁇ g/mL). ) is a graph analyzing the cytotoxicity and antimyostatin activity according to the treatment.
  • Figure 2 (b) shows the treatment of CAGA 12 -Luc-expressing 293T cell line with recombinant myostatin and various concentrations of fresh wild chives (flower, leaf, fruit or herb) extract (5, 20, 100 ⁇ g/mL). It is a graph analyzing the cytotoxicity and antimyostatin activity according to FIG.
  • Figure 3 is a photograph of analyzing the expression level of SMAD2 phosphorylated protein according to the treatment of HepG2 cell line with recombinant myostatin and various concentrations of extract (3.13, 6.25, 12.5, 25 ⁇ g/mL).
  • the present inventors While searching for a natural ingredient effective for muscle strength enhancement as an antimyostatin material, the present inventors found that, among Rosa plants, the extract of rosa genus sarcopenia inhibits myostatin activity without cytotoxicity, thereby strengthening muscle strength or preventing sarcopenia. Or it was confirmed that it can be effectively used for treatment, and the present invention was completed.
  • muscle weakness-related disease or sarcopenia refers to myostatin-related disease, which occurs in various catabolic conditions caused by hormonal imbalance, severe injury, sepsis, cancer, and aging. Loss of size and mass of cells and muscle tissue.
  • the diseases associated with muscle weakness include muscular atrophy, myasthenia gravis, muscular dystrophy, myasthenia gravis, hypotonia, muscle weakness, muscle degeneration, muscular dystrophy, amyotrophic lateral sclerosis, myasthenia gravis, and hypotonia. It may be one or more selected from the group consisting of (atony) and cachexia.
  • Prevention in the present specification refers to all actions that delay the onset of muscle weakness-related diseases or sarcopenia by administration of the composition of the present invention, and “treatment” and “improvement” are related to muscle weakness by administration of the composition of the present invention. Any action that improves or beneficially alters the symptoms of a disease or sarcopenia.
  • composition for preventing or treating muscle weakness-related diseases or sarcopenia for preventing or treating muscle weakness-related diseases or sarcopenia
  • the present invention provides a pharmaceutical composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia, containing an extract, a fraction thereof, or a fermented product thereof as an active ingredient.
  • the pharmaceutical composition for the prevention or treatment of muscle weakness-related diseases or sarcopenia contains, as an active ingredient, an extract, a fraction thereof, or a fermented product thereof.
  • the live hot tree belongs to a perennial plant belonging to the Rosaceae (Rosaceae) genus (Rosa), and is distributed in the Korean Peninsula, China, and Far East Russia, etc., and ornamental, wheat, spices, and fruits are edible.
  • Rosaceae Rosaceae
  • Rosa Rosaceae
  • ornamental, wheat, spices, and fruits are edible.
  • the above-mentioned green fever tree is effective in strengthening muscle strength.
  • the raw heat fir tree extract can be extracted from any one or more parts selected from the group consisting of leaves, stems, flowers, fruits and outposts of the fresh heat fir tree, and in the group consisting of flowers, leaves and It is preferable to extract any one or more selected sites, but is not limited thereto. At this time, the corresponding site may be in a dry state.
  • the extract of the raw heat fir tree can be extracted using water, an organic solvent, or a mixture thereof as a solvent, and extracting using water, C1 to C4 lower alcohol, or a mixture thereof as a solvent Preferred, but not limited thereto.
  • the extract of the fresh wild cypress tree is preferably extracted by adding water, an organic solvent or a mixture thereof 5 to 50 times the weight of the fresh cypress tree, and 10 to 30 times the weight of the dried cypress tree. It is more preferable to extract by adding pears, but is not limited thereto.
  • the extraction temperature is preferably 50 ° C to 150 ° C, more preferably 70 ° C to 120 ° C, but is not limited thereto.
  • the extraction time is preferably 1 hour to 20 hours, more preferably 3 hours to 10 hours, but is not limited thereto.
  • a cold needle extraction, ultrasonic extraction, or reflux cooling extraction method may all be used, but is not limited thereto.
  • the number of extractions is preferably 1 to 5 times, and more preferably repeated extraction 2 to 3 times, but is not limited thereto.
  • the extract of the wild genus japonica may be diluted, concentrated, or purified and dried after being diluted or concentrated.
  • a fraction of the natural cypress tree extract can be selected.
  • a polar solvent fraction and a non-polar solvent fraction may be respectively obtained by fractionation using a non-polar solvent such as ethyl acetate.
  • the fractions of the extract of the wild red ginseng plant may be separated by various chromatographies.
  • a fermented product of the natural wild plant extract may be selected instead of the natural plant extract, and the fermented product of the natural plant extract may be obtained by inoculating the plant extract with various lactic acid bacteria and then fermenting it.
  • the extract, a fraction thereof, or a fermented product thereof can inhibit myostatin activity without cytotoxicity, and also, by interfering with myostatin signal transduction, a signal that proceeds into cells (expression of SMAD2 phosphorylated protein) ) By inhibiting, it can have a muscle strengthening effect.
  • the muscle strength strengthening effect of the extract, a fraction thereof, or a fermented product thereof is far superior to that of a similar plant belonging to the genus Rosa. For example, as a similar plant of the live fever tree, white live fever ( Rosa davurica for.alba ), long live fever ( Rosa davurica var. There are limitations in which reinforcement is not effective.
  • the concentration of the wild chives extract, its fraction or its fermented product may be 0.1 ⁇ g/mL to 100 ⁇ g/mL, preferably 0.1 ⁇ g/mL to 25 ⁇ g/mL, and 0.4 ⁇ g/mL to 25 ⁇ g/mL It is more preferable, but is not limited thereto. At this time, if the concentration is less than the above range, there is a problem that the effect of inhibiting myostatin activity is insignificant and is not effective for muscle strength or sarcopenia, and if the concentration exceeds the above range, toxicity concerns including cytotoxicity This can be.
  • compositions may be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively.
  • Suitable carriers, excipients or diluents commonly used in the preparation of pharmaceutical compositions may be included.
  • carrier or excipient or diluent examples include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, undecided and various compounds or mixtures including vaginal cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
  • diluents or excipients such as commonly used fillers, weighting agents, binders, wetting agents, disintegrants, and surfactants.
  • a solid preparation for oral administration may be prepared by mixing at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the extract of the wild chive plant.
  • excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, etc.
  • lubricants such as magnesium stearate and talc may also be used.
  • Liquid preparations for oral administration include suspensions, solutions for oral use, emulsions, syrups, etc., and various excipients such as wetting agents, sweeteners, aromatics, preservatives, etc. may be included in addition to water and liquid paraffin, which are commonly used simple diluents. .
  • Formulations for parenteral administration include sterilized aqueous solutions, water-insoluble agents, suspensions, emulsions, freeze-dried preparations, and suppositories.
  • Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions.
  • injectable esters such as ethyl oleate
  • a base for suppositories witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like can be used.
  • the preferred dosage of the pharmaceutical composition varies depending on the patient's condition, body weight, disease severity, drug type, administration route and period, but can be appropriately selected by those skilled in the art. However, for desirable effects, it may be administered at 0.0001 to 2,000 mg/kg per day, preferably 0.001 to 2,000 mg/kg. Administration may be administered once a day or divided into several times. However, the scope of the present invention is not limited by the dosage.
  • the pharmaceutical composition may be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
  • composition for preventing or treating muscle weakness-related diseases or sarcopenia for preventing or treating muscle weakness-related diseases or sarcopenia
  • the present invention provides a health functional food composition or food composition for strengthening muscle strength or preventing or improving sarcopenia, which contains an extract of genus japonica, a fraction thereof, or a fermented product thereof as an active ingredient.
  • the health functional food composition or food composition for strengthening muscle strength or for preventing or improving sarcopenia contains as an active ingredient an extract of wild chives, a fraction thereof, or a fermented product thereof. Since the fermented product has been described above, redundant description will be omitted.
  • the concentration of the wild chives extract, its fraction or its fermented product may be 0.1 ⁇ g/mL to 100 ⁇ g/mL, preferably 0.1 ⁇ g/mL to 25 ⁇ g/mL, and 0.4 ⁇ g/mL to 25 ⁇ g/mL It is more preferable, but is not limited thereto. At this time, if the concentration is less than the above range, there is a problem that the effect of inhibiting myostatin activity is insignificant and is not effective for muscle strength or sarcopenia, and if the concentration exceeds the above range, toxicity concerns including cytotoxicity This can be.
  • the extract of the health functional food or food additives when using the extract of the health functional food or food additives, it may be added as it is or used together with other foods or food ingredients, and appropriately according to a conventional method can be used.
  • the mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health, or treatment.
  • the formulation of the health functional food composition or food composition may be in the form of powder, granules, pills, tablets, capsules, as well as general food or beverage forms.
  • the type of food is not particularly limited, and examples of food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, etc., and may include all foods in a conventional sense.
  • the extract of the genus japonica may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on 100 parts by weight of the raw material.
  • the amount may be less than the above range, and since the present invention uses natural substances, there is no problem in terms of safety, so the amount above the above range can also be used as
  • Beverages of the health functional food composition or food composition may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages.
  • the aforementioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used.
  • the ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage.
  • the health functional food composition or food composition is various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, It may contain glycerin, alcohol, and carbonation agents used in carbonated beverages.
  • the health functional food composition or food composition may contain fruit flesh for the production of natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not limited, but is generally selected from the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food composition or food composition.
  • the present invention relates to a composition for preventing or treating muscle weakness-related diseases or sarcopenia, which contains as an active ingredient an extract, a fraction thereof, or a fermented product thereof as a natural ingredient. , Since it can inhibit myostatin activity, it can be effectively used for strengthening muscle strength or preventing or treating sarcopenia.
  • the present invention provides a use for the use of the extract, a fraction thereof, or a fermented product thereof in a pharmaceutical composition for preventing or treating muscle weakness-related diseases or sarcopenia.
  • the present invention provides a use for the use of the extract, a fraction thereof, or a fermented product thereof in a health functional food composition or food composition for strengthening muscle strength or preventing or improving sarcopenia.
  • the present invention provides a method for preventing or treating muscle weakness-related diseases or sarcopenia, comprising the step of administering to a subject an extract, a fraction thereof, or a fermented product thereof.
  • the present invention provides a method for strengthening muscle strength or a method for preventing or improving sarcopenia, comprising the step of administering to a subject an extract, a fraction thereof, or a fermented product thereof.
  • subject means a subject in need of treatment of a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat, horse, cow, etc. of mammals.
  • Table 1 shows the yield of extracts of wild genus japonica (flowers, leaves or fruits) by classification by extraction part or extraction solvent.
  • 293T cell line expressing CAGA 12 -Luc at 5 ⁇ 10 4 cells/100 uL per well was prepared in a 96-well plate using 10% fetal bovine serum DMEM medium without phenol red. Divided and cultured at 5% CO 2 , 37°C. After 24 hours, the serum-free DMEM medium without phenol red was mixed with the extract of the wild genus japonica (herbica) and treated by concentration (12.5, 25, 50, 100 ⁇ g/mL), but the control group (Vehicle) was CAGA 12 -Luc The 293T cell line expressing the was treated with only the solvent and cultured at 5% CO 2 and 37°C.
  • the concentration of the extract of wild genus japonica (whole plant) in the 293T cell line in which CAGA 12 -Luc is expressed is less than 100 ⁇ g / mL, preferably less than 25 ⁇ g / mL, cytotoxicity is shown. It is confirmed that no
  • the 293T cell line expressing CAGA 12 -Luc at 5 ⁇ 10 4 cells/100 uL per well was dispensed into a 96 well plate using 10% fetal calf serum DMEM medium without phenol red, 5% CO 2 , 37 °C cultured in.
  • the recombinant myostatin (10 nM) and the extract of wild genus japonica (herbica) were mixed in a serum-free DMEM medium without phenol red, and the concentrations (0.2, 0.4, 0.8, 1.6, 3.1, 6.3, 12.5, 25 ug/mL), but as a control group (Vehicle), CAGA 12 -Luc-expressing 293T cell line was treated with only the solvent and cultured at 5% CO 2 and 37°C.
  • the concentration of the wild genus japonica extract in the 293T cell line expressing CAGA 12 -Luc was 25 ⁇ g/mL or less, specifically, at 0.2 ⁇ g/mL to 25 ⁇ g/mL. All of them were confirmed to show no cytotoxicity with a cell viability of 80% or more. In addition, it is confirmed that the extract of wild lentils has a tendency to suppress myostatin activity in a concentration-dependent manner.
  • the extract of the wild plant (flower, leaf, fruit or plant) by concentration (0.2, 0.4, 0.8, 1.6, 3.1, 6.3, 12.5, 25 ⁇ g/mL)
  • the extract of the plant were treated at different concentrations (5, 20, 100 ⁇ g/mL) and repeated experiments were performed.
  • the extract of wild genus japonica tends to suppress myostatin activity in a concentration-dependent manner.
  • myostatin activity was strongly inhibited at the same concentration in the case of the extract of the wild cypress (flower, leaf and herb) extract, compared to the extract of the genus cypress (fruit).
  • myostatin activity was strongly inhibited at the same concentration when 30-70% ethanol was used as the extraction solvent, compared to the case where water was used as the extraction solvent.
  • 5.0 ⁇ 10 5 cells of the HepG2 cell line were dispensed per well in a 12-well plate using 10% fetal bovine serum DMEM medium and cultured at 5% CO 2 and 37°C. After 24 hours, the medium was replaced with serum-free DMEM medium and cultured for 4 hours at 5% CO 2 and 37°C.
  • a bloodless DMEM medium was mixed with recombinant myostatin (10 nM) and the extract of wild genus chinensis, and treated by concentration (3.13, 6.25, 12.5, 25 ⁇ g/mL), 5% CO 2 , at 37°C for 30 minutes. cultured.
  • the cultured cells were treated with a mixture of EDTA and a phosphatase inhibitor in RIPA buffer after washing with PBS buffer to remove the culture medium. After scraping the cells with a scraper, the cells were recovered. After the pellet was homogenized and reacted on ice for 30 minutes, the supernatant obtained by centrifugation at 4° C. and 13,000 rpm for 10 minutes was quantified by BCA. After electrophoresis, the cells were blocked for 1 hour at room temperature and reacted with primary antibodies (pSMAD2, SMAD2, ⁇ -actin) at 4°C. After 24 hours, the secondary antibody was reacted at room temperature for 1 hour, and the protein expression level was measured using Odyssey CLX.
  • primary antibodies pSMAD2, SMAD2, ⁇ -actin

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Abstract

La présente invention concerne une composition pour prévenir ou traiter des maladies associées à une faiblesse musculaire ou une sarcopénie, contenant, en tant que principe actif, un extrait de Rosa davurica Pallas, une fraction de celui-ci ou un produit fermenté de celui-ci.
PCT/KR2022/014275 2021-09-23 2022-09-23 Composition pour prévenir ou traiter des maladies associées à une faiblesse musculaire ou une sarcopénie, contenant un extrait de rosa davurica pallas en tant que principe actif WO2023048504A1 (fr)

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KIM JOON-BUM, CHENG-BI CUI, DEUK-SIK LEE, SEUNG-SHI HAM: "Studies on Biological Activity of Leaves from Korean Rosa davurica Pall.", KOREAN JOURNAL OF FOOD PRESERVATION, KOREAN INTELLECTUAL PROPERTY OFFICE, vol. 11, no. 1, 1 March 2004 (2004-03-01), pages 100 - 105, XP093053459, ISSN: 1738-7248 *

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