WO2006115321A1 - Composition contenant un extrait de salvia miltiorrhiza bge presentant une activite facilitante pour la prevention ou le traitement des maladies circulatoires - Google Patents

Composition contenant un extrait de salvia miltiorrhiza bge presentant une activite facilitante pour la prevention ou le traitement des maladies circulatoires Download PDF

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Publication number
WO2006115321A1
WO2006115321A1 PCT/KR2006/000026 KR2006000026W WO2006115321A1 WO 2006115321 A1 WO2006115321 A1 WO 2006115321A1 KR 2006000026 W KR2006000026 W KR 2006000026W WO 2006115321 A1 WO2006115321 A1 WO 2006115321A1
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Prior art keywords
extract
polar solvent
mmp
salvia miltiorrhiza
solvent soluble
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PCT/KR2006/000026
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English (en)
Inventor
Hyun Wook Chang
Jong-Keun Son
Seung Ho Lee
Kun Ho Son
Kyung Soo Nam
Cheorl Ho Kim
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Industry Academic Cooperation Foundation, Yeungnam University
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Publication of WO2006115321A1 publication Critical patent/WO2006115321A1/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)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/537—Salvia (sage)
    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to an extract of Salvia miltiorrhiza B GE. showing potent inhibiting effect onmatrix metalloproteinase-9 activity and migration of TNF- ⁇ -induced human aortic smooth muscle cells and a composition comprising the same having preventing and treating arteriosclerosis.
  • vascular smooth muscle cells may play a key role in the development of intimal thickening after arterial wall injury or in atherosclerosis (Ross, R., New England Journal of Medicine 314. pp488-500. 1986). VSMC in the media has low mitogenic activity. During the early stages of arterial wall injury or atherosclerosis, aortic smooth muscle cells may undergo transition from a contractile to a synthetic phenotype and begin proliferating in response to various growth factors, causing intimal thickening of the arterial walls (Chamley-Campbell, et al., Physiological Review 59_, ppl-61.
  • VSMC synthesizes important components of the extracellular matrix, including collagens, elastin, and proteoglycans (Galis et al., Circulation Research 75, ppl81-189. 1994).
  • An imbalance between the accumulation and degradation of extracellular matrix may be crucial in the development of intimal thickening that forms after vascular wall interventions (Strauss et al., Circulation Research 75, pp650-658. 1994).
  • MMPs Matrix metalloproteinases
  • ECM extracellular matrix
  • MMPs comprise three main groups: the interstitial collagenases (MMP-I), the type IV collagenases or gelatinases (MMP-2 and -9), and the stromelysins (MMP-3) (Woessner, 1991 Woessner, J.F.Jr., FASEB Journal 5, pp2145-2154. 1991; Chung et al., FASEB Journal 18QOl ppl 123-1125. 2004a).
  • MMPs interstitial collagenases
  • MMP-2 and -9 the type IV collagenases or gelatinases
  • MMP-3 stromelysins
  • gelatinases degrade denatured interstitial collagens, native basement-membrane collagens
  • the expression of MMP-9 has been implicated in the progression of atherosclerotic lesions (Newby and Zaltsman, Journal of Pathology 190. pp300-309.
  • MMP-9 is critical for the development of arterial lesions by regulating both VSMC migration and proliferation.
  • TNF- ⁇ tumor necrosis factor- ⁇
  • ERK1/2 mediates TNF- ⁇ induced MMP-9 expression in VSMC via the regulation of NF-kB and AP-I (Moon, S. K., et al., Biochemical and Biophysical Research Communication 301. ppl069-1078. 2003; Moon, S.K., et al., Archives of Biochemistry and Biophysics 418, pp39-48. 2003b).
  • BB94 bathimastat
  • BB2516 marimastat
  • BB94 has a structure that mimics collagen and facilitates chelation of the zinc ion in the active site of the MMP molecule, thereby causing an inactive protease.
  • substances of this type have been used to study the importance of extracellular matrix remodeling in various types of diseases, including rheumatoid arthritis, cancer invasion and cardiovascular diseases.
  • SM (Labiatae) contains phenolic compounds that are effective in protecting liver microsomes, hepatocytes, and erythrocytes against oxidative damage (Li et al., Journal of Asian Natural Products Research 414), pp271-280, 2002; Liu et al., Liver 21 f 6 * ). pp384-390, 2001) and several active components such as tanshinones, D(+)3,4-dihydroxyphenol lactic acid, protocatechuic aldehyde, salvianolic acids (A, B, C, D, E, F) and rosmarinic acid have been isolated and identified therefrom (Li, L.N., Journal of Chinese Pharmaceutical Science 6, pp57-64.1997).
  • active components such as tanshinones, D(+)3,4-dihydroxyphenol lactic acid, protocatechuic aldehyde, salvianolic acids (A, B, C, D, E, F) and rosmarinic acid have been isolated and identified therefrom (
  • a pharmaceutical composition comprising polar solvent soluble or non-polar solvent soluble extract of Salvia miltiorrhiza BGE as an active ingredients for the treatment and prevention of blood circulatory system disorder caused by MMP hyper-activation.
  • polar solvent soluble or non-polar solvent soluble extract of Salvia miltiorrhiza BGE can be fractionated from crude extract prepared by the steps consisting of: extracting the rhizoma, root, or herb of plant material with water, lower alcohols such as methanol, ethanol, preferably methanol and the like, or the mixtures thereof.
  • polar solvent soluble extract can be prepared by extracting above crude extract with polar solvent, for example, water, lower alcohol such as methanol, ethanol, butanol preferably butanol and the like, or the mixtures thereof.
  • polar solvent for example, water, lower alcohol such as methanol, ethanol, butanol preferably butanol and the like, or the mixtures thereof.
  • non-polar solvent soluble extract can be prepared by extracting above crude extract with non-polar solvent, for example, hexane, ethyl acetate or dichloromethane, preferably ethyl acetate.
  • non-polar solvent for example, hexane, ethyl acetate or dichloromethane, preferably ethyl acetate.
  • Above described blood circulatory system disorder comprises atherosclerosis, ischemic myocardial infarction, angina pectoris, hypertension, stroke, hyperlipemia, anemia, migraine, arrhythmia, apoplexy, angioma, hemangiofibroma, vascular deformity, angiostenosis and the like, preferably, atherosclerosis.
  • Salvia japonica or Salvia plebia Salvia japonica or Salvia plebia.
  • composition of the present invention can contain about 0.01 ⁇
  • the health food of the present invention comprises above extracts as 0.01 to 80 %, preferably 1 to 50 % by weight based on the total weight of the composition.
  • Above health food can be contained in health food, health beverage etc, and may be used as powder, granule, tablet, chewing tablet, capsule, beverage etc.
  • the inventive crude extract of Salvia miltiorrhiza BGE can be prepared by follows;
  • the rhizoma of Salvia miltiorrhiza BGE is dried, cut, crushed and mixed with 5 to 25-fold, preferably, approximately 10 fold volume of distilled water, lower alcohols such as methanol, ethanol, butanol and the like, or the mixtures thereof, preferably methanol; the solution is treated with hot water at the temperature ranging from 20 to 100°C, preferably from 60 to 100°C, for the period ranging from 15mins to 24 hours with extraction method by the extraction with hot water, cold water, reflux extraction, or ultra-sonication extraction with 1 to 5 times, preferably 2 to 3 times, consecutively; the residue is filtered to obtain the supernatant to be concentrated with rotary evaporator, at the temperature ranging from 20 to 100°C, preferably from 50 to 70°C and then dried by vacuum freeze-drying, hot air-drying or spray drying to obtain dried crude extract powder of Salvia miltiorrhiza BGE which can be soluble in water, lower alcohols, or the mixtures
  • polar solvent soluble and non-polar solvent soluble extract of present invention can be prepared by following procedure; the crude extract prepared by above step, is suspended in water, and then is mixed with 1 to 100-fold, preferably, 1 to 5-fold volume of non polar solvent such as ethyl acetate, chloroform, hexane and the like; the non-polar solvent soluble layer is collected to obtain non-polar solvent soluble extract of the present invention and remaining polar solvent soluble layer is collected to obtain polar solvent soluble extract of the present invention which is soluble in water, lower alcohols, or the mixtures thereof.
  • non polar solvent such as ethyl acetate, chloroform, hexane and the like
  • a pharmaceutical composition comprising the polar solvent soluble or non-polar solvent soluble extract of Salvia miltiorrhiza prepared by above preparation method for the treatment and prevention of blood circulatory disease caused by MMP hyper-activation in human or mammal as active ingredients.
  • the inventive composition for treating and preventing blood circulatory disease may comprises above extracts as 0.01 ⁇ 50 % by weight based on the total weight of the composition.
  • inventive composition may additionally comprise conventional carrier, adjuvants or diluents in accordance with a using method well known in the art. It is preferable that said carrier is used as appropriate substance according to the usage and application method, but it is not limited. Appropriate diluents are listed in the written text of Remington's Pharmaceutical Science (Mack Publishing co, Easton PA).
  • composition according to the present invention can be provided as a pharmaceutical composition containing pharmaceutically acceptable carriers, adjuvants or diluents, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil.
  • pharmaceutically acceptable carriers e.g., lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl
  • the formulations may additionally include fillers, anti-agglutinating agents, lubricating agents, wetting agents, flavoring agents, emulsifiers, preservatives and the like.
  • the compositions of the invention may be formulated so as to provide quick, sustained or delayed release of the active ingredient after their administration to a patient by employing any of the procedures well known in the art.
  • compositions of the present invention can be dissolved in oils, propylene glycol or other solvents that are commonly used to produce an injection.
  • suitable examples of the carriers include physiological saline, polyethylene glycol, ethanol, vegetable oils, isopropyl myristate, etc., but are not limited to them.
  • the extract of the present invention can be formulated in the form of ointments and creams.
  • compositions containing present composition may be prepared in any form, such as oral dosage form (powder, tablet, capsule, soft capsule, aqueous medicine, syrup, elixirs pill, powder, sachet, granule), or topical preparation (cream, ointment, lotion, gel, balm, patch, paste, spray solution, aerosol and the like), or injectable preparation (solution, suspension, emulsion).
  • composition of the present invention in pharmaceutical dosage forms may be used in the form of their pharmaceutically acceptable salts, and also may be used alone or in appropriate association, as well as in combination with other pharmaceutically active compounds.
  • the desirable dose of the inventive extract or composition varies depending on the condition and the weight of the subject, severity, drug form, route and period of administration, and may be chosen by those skilled in the art. However, in order to obtain desirable effects, it is generally recommended to administer at the amount ranging 10 g/kg, preferably, 1 to 3 g/kg by weight/day of the inventive extract or compounds of the present invention. The dose may be administered in single or divided into several times per day. In terms of composition, the amount of inventive extract should be present between 0.01 to 50% by weight, preferably 0.5 to 40% by weight based on the total weight of the composition.
  • composition of present invention can be administered to a subject animal such as mammals (rat, mouse, domestic animals or human) via various routes. All modes of administration are contemplated, for example, administration can be made orally, rectally or by intravenous, intramuscular, subcutaneous, intracutaneous, intrathecal, epidural or intracerebroventricular injection.
  • the present invention provide a composition of the health food beverage for the prevention and improvement of blood circulatory disease adding above described extracts 0.01 to 80 % by weight, amino acids 0.001 to 5 % by weight, vitamins 0.001 to 2 % by weight, sugars 0.001 to 20 % by weight, organic acids 0.001 to 10 % by weight, sweetener and flavors of proper amount.
  • examples of addable food comprising above extracts of the present invention are various food, beverage, gum, vitamin complex, health improving food and the like, and can be used as power, granule, tablet, chewing tablet, capsule or beverage etc.
  • the extract of the present invention will be able to prevent and improve blood circulatory disease by way of adding to child and infant food, such as modified milk powder, modified milk powder for growth period, modified food for growth period.
  • composition therein can be added to food, additive or beverage, wherein, the amount of above described extract in food or beverage may generally range from about 0.1 to 80w/w %, preferably 1 to 50 w/w % of total weight of food for the health food composition and 1 to 30 g, preferably 3 to 10 g on the ratio of IOOD of the health beverage composition.
  • the health beverage composition of present invention contains above described extract as an essential component in the indicated ratio
  • the other component can be various deodorant or natural carbohydrate etc such as conventional beverage.
  • natural carbohydrate are monosaccharide such as glucose, fructose etc; disaccharide such as maltose, sucrose etc; conventional sugar such as dextrin, cy- clodextrin; and sugar alcohol such as xylitol, and erythritol etc.
  • natural deodorant such as taumatin, stevia extract such as levaudioside A, glycyrrhizin et al., and synthetic deodorant such as saccharin, aspartam et al.
  • the amount of above described natural carbohydrate is generally ranges from about 1 to 20 g, preferably 5 to 12 g in the ratio of 100 D of present beverage composition.
  • the other components than aforementioned composition are various nutrients, a vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al.
  • the other component than aforementioned ones may be fruit juice for preparing natural fruit juice, fruit juice beverage and vegetable beverage, wherein the component can be used independently or in combination.
  • the ratio of the components is not so important but is generally range from about 0 to 20 w/w % per 100 w/w % present composition.
  • Examples of addable food comprising aforement ioned extract therein are various food, beverage, gum, vitamin complex, health improving food and the like.
  • the inventive composition may additionally comprise one or more than one of organic acid, such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid; phosphate, such as phosphate, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate; natural anti-oxidants, such as polyphenol, catechin, ⁇ - tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.
  • organic acid such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid
  • phosphate such as phosphate, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate
  • natural anti-oxidants such as polyphenol, catechin, ⁇ - tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.
  • the above extract of Salvia miltiorrhiza BGE may be 20 to 90 % high concentrated liquid, power, or granule type.
  • the above extract of Salvia miltiorrhiza BGE can comprise additionally one or more than one of lactose, casein, dextrose, glucose, sucrose and sorbitol.
  • Inventive extract of the present invention have no toxicity and adverse effect therefore; they can be used with safe.
  • a pharmaceutical composition comprising the polar solvent soluble or non-polar solvent soluble extract of Salvia miltiorrhiza prepared by above preparation method for the treatment and prevention of blood circulatory disease caused by MMP hyper-activation in human or mammal as active ingredients.
  • Fig. 1 shows the effect of the extract of Salvia miltiorrhiza BGE on cell proliferation
  • Fig. 2 shows the photographs of HASMC cells treated with control group (A), 100
  • Fig. 3 represents the MMP-9 gelatinolytic effect on the cultured conditioned media of HASMC treated with TNF- ⁇ (100 ng/ml); lane 1 treated with control group, lane 2 with buffer and 50 D of 70% EtOH soluble extract, lane 3 with buffer and 50 D of chloroform soluble extract, lane 4 with buffer and 50 D of ethylacetate soluble extract, lane 5 with buffer and 50 D of butanol soluble extract, lane 6 with buffer and 50 D of water soluble extract;
  • Fig. 4 represents the inhibitory effects on the MMP-9 gelatinolytic activity of various concentration of EtOAc soluble extract (0, 10, 20, 30, 40, 50, 70, 100 and 500 D/ml) and zymographs performed by controlled medium obtained from HASMC cells in the presence of TNF- ⁇ or not;
  • Fig. 5 presents the inhibitory effect of ethylacetate soluble extract on the migration of VSMC cells
  • Fig. A shows the migrated cells excluded TNF- ⁇ or inventive extract (a)
  • the cells treated with 100 ng/ml of TNF- ⁇ (b)
  • the cells treated with TNF- ⁇ and 100 D/ml of inventive extract (c)
  • the cells treated with TNF- ⁇ and 200 D/ml of inventive extract (d)
  • Fig. B shows total number of migrated cells stained with hematoxylin and eosin;
  • Fig 6 depicts the inhibitory effect of the inventive extract compared with previously known MMP inhibitors on MMP-9;
  • A cultured medium treated with DMSO, EDC and 50 D/ml of inventive extract,
  • B the intensity of the bands estimated with Gel-Print System transformed into expressed mean+SE value in zymogram studies.
  • (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG) were obtained from Sigma Chemical Co. (MO, USA). The drugs were dissolved in dimethyl sulfoxide (DMSO) at a concentration of 40 mM. Recombinant human TNF- ⁇ and BB94 was obtained from R&D systems (MN, USA) and British Biotechnology (Oxford, U.K.), respectively.
  • DMSO dimethyl sulfoxide
  • MASMC Human aortic smooth muscle cells
  • HASMC were cultured in smooth muscle cell growth medium-2 containing 10%
  • FBS 2 ng/ml human basic fibroblast growth factor, 0.5 ng/ml human epidermal growth factor, 50 D/ml gentamicin, 50 D/ml amphotericin-B, and 5 D/ml bovine insulin.
  • early passage HASMC were grown to 80-90% confluence and made quiescent by serum starvation (0.1% FBS) for at least 24 hrs.
  • the serum-free medium contained secreted proteins such as MMP-9. The amount of secreted proteins in the conditioned media were estimated and quantified by cell numbers.
  • XTT reaction solution sodium 3'-[l-(phenyl-aminocarbonyl)-3,4-tetrazolium] - bis(4-methoxy-6-nitro) benzenesulfonic acid hydrate and N-methyl dibenzopyrazine methyl sulfate; mixed in proportion 50:1) was added to the wells.
  • the optical density was read at 490 nm wavelength in an ELISA plate reader after 4 hrs incubation of the plates with XTT in an incubator (37°C and 5% CO + 95% air). All determinations were confirmed using replication in at least three identical experiments. The data were shown mean+SE as percent of control.
  • the inventive extract prepared in Example 1 showed weak cytotoxic effect on HASMC cells in a dose dependent manner (IC > 500 D/ml), which is not so harmful to human body.
  • the CH 2 Cl 2 fraction, water extract and 70% EtOH extract exhibited weak cytotoxic activity (IC of > 100 D/ml, 90 D/ml, and >100 D/ml, re-
  • the gels were soaked in 0.25% Triton X-100 (2x30 min) at room temperature and rinsed in NanoPure water.
  • Triton X-100 2x30 min
  • the methanol extract and its fractions were freshly solubilized in the Tris buffer used for developing the zymogram; the gel slab was cut into slices corresponding to the lanes and then put in different tanks containing the stated concentrations of fractions.
  • the gel containing gelatin was incubated at 37°C for 20 h in the incubation buffer containing 50 mM Tris-HCl (pH 7.6), 20 mM NaCl, 5 mM CaCl 2 and 0.02% Brij-58 with or without 0, 1, 10, 50, 100, 500, 1000 D/ml of fraction or extract.
  • the gel was then stained for 15-30 min in 0.1% (w/v) Coomassie blue R-250 in 30% methanol and 10% acetic acid, and destained in the same solution without the Coomassie blue dye.
  • SM is a strong inhibitor of MMP-9.
  • the inhibitory effect against MMP-9 activity of SM was compared with established MMP inhibitors such as BB 94, catechin and its derivates, which are present abundantly in green tea.
  • MMP inhibitors such as BB 94, catechin and its derivates, which are present abundantly in green tea.
  • the inhibitory effect of SM against gelatinolytic activity of MMP-9 was higher than that of EGC, but lower than that of EGCG and BB94.
  • Matrigel-coated filter inserts (8 D pore size) that fit into 24-well invasion chambers were obtained from Becton-Dickinson (NJ, USA).
  • HASMC (5 X 10 cells/ well) to be tested for invasion were detached from the tissue culture plates, washed, re- suspended in conditioned medium collected from TNF- ⁇ treated HASMC for 24 h, and then added to the upper compartment of the invasion chamber in various concentration of SM (0, 50, 100, 250 and 500 D/ml). Five hundred ml of same conditioned medium was added to the lower compartment of the invasion chamber. Cells without TNF- ⁇ treated conditioned medium served as control. The Matrigel invasion chambers were incubated at 37°C for 24 hrs in 5% CO 2.
  • the filter inserts were removed from the wells, and the cells on the upper side of the filter were removed using cotton swabs.
  • the filters were fixed, mounted, and stained according to the manufacturer's instructions. The cells that invaded through the Matrigel and were located on the underside of the filter were counted. Three to five invasion chambers were used per condition. The values obtained were calculated by averaging the total number of cells from three filters.
  • VSMC were resuspended in conditioned medium (5x10 4 cells/200 ml), added to the upper components of the matrigel invasion chamber supplemented with various concentration of EtOAc fraction (0, 50, 100, 250 and 500 D/ml) and incubated for 24 h at 37°C and 5% CO 2.
  • Powder preparation was prepared by mixing above components and filling sealed package.
  • Tablet preparation was prepared by mixing above components and entabletting.
  • Tablet preparation was prepared by mixing above components and filling gelatin capsule by conventional gelatin preparation method.
  • Injection preparation was prepared by dissolving active component, controlling pH to about 7.5 and then filling all the components in 2ml ample and sterilizing by conventional injection preparation method.
  • Liquid preparation was prepared by dissolving active component, filling all the components and sterilizing by conventional liquid preparation method.
  • Vitamin mixture optimum amount
  • Vitamin B 0.5mg
  • Vitamin B 0.2mg
  • Vitamin C lOmg
  • Health beverage preparation was prepared by dissolving active component, mixing, stirred at 85°C for 1 hour, filtered and then filling all the components in IOOOD ample and sterilizing by conventional health beverage preparation method.
  • the polar solvent soluble extract and non- polar solvent soluble extract inhibit MMP-9 inhibition activity, the cytotoxicity of HASMC cell etc,therefore, it can be used as the therapeutics or health food for treating and preventing blood circulatory disorder such as atherosclerosis without adverse action.

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Abstract

L'invention concerne une composition contenant un extrait par solvant soluble polaire ou par solvant soluble non polaire de Salvia miltiorrhiza, destinée à la prévention et au traitement des troubles du système circulatoire. Les extraits de Salvia miltiorrhiza ont un puissant effet inhibiteur sur l'activité protéolytique des métalloprotéases matricielles MMP-9 et sur la migration des cellules des muscles lisses vasculaires (MLV) et conviennent par conséquent comme agents thérapeutiques ou compléments alimentaires pour le traitement et la prévention des troubles du système circulatoire causés l'hyperactivation des MMP.
PCT/KR2006/000026 2005-04-25 2006-01-04 Composition contenant un extrait de salvia miltiorrhiza bge presentant une activite facilitante pour la prevention ou le traitement des maladies circulatoires WO2006115321A1 (fr)

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KR20050034153 2005-04-25
KR10-2005-0034153 2005-04-25

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Cited By (5)

* Cited by examiner, † Cited by third party
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ITMI20101833A1 (it) * 2010-10-08 2012-04-09 Velleja Res Srl Metodo per la preparazione di estratti totali da piante medicinali o alimentari mediante estrazione frazionata
CN102526234A (zh) * 2011-12-03 2012-07-04 范文忠 一种治疗中风的药物及其制备方法
CN102552468A (zh) * 2011-12-28 2012-07-11 蚌埠丰原涂山制药有限公司 一种治疗高血脂的中药组合物
CN107519360A (zh) * 2017-09-21 2017-12-29 张昕 一种防治高血压的中药枕

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CN102441117A (zh) * 2011-11-30 2012-05-09 宋爱民 治疗血管性头痛的中药制剂
CN102920837A (zh) * 2012-10-31 2013-02-13 侯明晓 一种治疗心律失常的中药及其制备工艺和应用
KR20200058900A (ko) 2018-11-20 2020-05-28 주식회사 뉴랜드올네이처 단삼(Salvia　miltiorrhiza) 추출물 또는 발효물을 포함하는 화장료 조성물 및 이의 용도
KR102244508B1 (ko) * 2019-05-17 2021-04-27 (주)유니베라 단삼 추출물 및 계피 추출물의 혼합물을 포함하는 혈관 노화로 인한 증상의 개선, 치료 또는 예방용 조성물
KR102218529B1 (ko) * 2019-07-17 2021-02-22 주식회사 리바이오 단삼 생물전환 추출물 및 홍삼 생물전환 추출물을 포함하는 심혈관계 질환 개선, 예방 또는 치료용 조성물
KR20210073679A (ko) 2019-12-10 2021-06-21 서울대학교산학협력단 설포라핀을 유효성분으로 함유하는 동맥경화 개선용 식품 조성물 및 동맥경화 치료 또는 예방용 약학 조성물

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ITMI20101833A1 (it) * 2010-10-08 2012-04-09 Velleja Res Srl Metodo per la preparazione di estratti totali da piante medicinali o alimentari mediante estrazione frazionata
CN102526234A (zh) * 2011-12-03 2012-07-04 范文忠 一种治疗中风的药物及其制备方法
CN102552468A (zh) * 2011-12-28 2012-07-11 蚌埠丰原涂山制药有限公司 一种治疗高血脂的中药组合物
CN107519360A (zh) * 2017-09-21 2017-12-29 张昕 一种防治高血压的中药枕

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