US20060078632A1 - Composition comprising bamboo extract and the compounds isolated therefrom showing treating and preventing activity for inflammatory and blood circulation disease - Google Patents

Composition comprising bamboo extract and the compounds isolated therefrom showing treating and preventing activity for inflammatory and blood circulation disease Download PDF

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US20060078632A1
US20060078632A1 US10/522,832 US52283205A US2006078632A1 US 20060078632 A1 US20060078632 A1 US 20060078632A1 US 52283205 A US52283205 A US 52283205A US 2006078632 A1 US2006078632 A1 US 2006078632A1
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Prior art keywords
extract
bamboo
pharmaceutical composition
sasa
polar solvent
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Inventor
Sung-Sick Woo
Tae Hyung Sung
Dong Seon Kim
Sun-Young Sung
Seon-Gil Do
Young Chul Lee
Jeong Bum Nam
Jong Ha Ryu
Kang Lee
Hee Sun Sung
Young Moon Heo
Mi-Sun Oh
Ji Nyeo Cho
Sookyoung Sung
Ju Yeon Lee
Tae Woo Kim
Ji Sook Song
Seoung Ho Lee
Mi Ran Kim
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Unigen Inc
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Unigen Inc
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Priority claimed from KR1020030019229A external-priority patent/KR100417243B1/ko
Priority claimed from KR1020030019230A external-priority patent/KR100417244B1/ko
Application filed by Unigen Inc filed Critical Unigen Inc
Assigned to UNIGEN, INC. reassignment UNIGEN, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: CHO, JI NYEO, DO, SEON-GIL, HEO, YOUNG MOON, JO, TAE HYUNG, KIM, DONG SEON, KIM, MI RAN, KIM, TAE WOO, LEE, JU YEON, LEE, KANG WOO, LEE, SEOUNG HO, LEE, YOUNG CHUL, NAM, JEONG BUM, OH, MI-SUN, RYU, JONG HA, SONG, JI SOOK, SUNG, HEE SUN, SUNG, SOOKYOUNG, SUNG, SUN-YOUNG, WOO, SUNG-SICK
Publication of US20060078632A1 publication Critical patent/US20060078632A1/en
Priority to US11/872,845 priority Critical patent/US7897182B2/en
Priority to US13/013,880 priority patent/US20110117224A1/en
Priority to US13/191,619 priority patent/US20110280975A1/en
Pending legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • 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
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • 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
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • 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
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • 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
    • 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
    • A61P9/12—Antihypertensives
    • 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 bamboo extract and the compound isolated therefrom showing treating and preventing activity for inflammatory and blood circulation disease.
  • Blood circulation disorder occurred by the blockage of blood flow caused by deposited cholesterol and increased thrombus on blood vessel, and lessened elastic force of blood vessel.
  • the representative symptoms of blood circulation disorder are benumbed feet or hand, crick of neck and shoulders, loss of memory, lethargy, loss of concentration, vertigo and chronic fatigue etc., which often give difficulty in normal living life.
  • Hyperlipemia as one example of blood circulation disorder, is a condition where the blood lipid parameters are elevated in the blood. This condition manifests an abnormally high concentration of fats in the blood.
  • the lipid component in the circulating blood is total cholesterol, low density lipoproteins, very low density lipoproteins or triglycerides.
  • Inflammation occurred by the invasion of outer contaminant, for example, bacteria, fungus, virus, various allergens inducing inflammatory response and a biophysical process against them.
  • the characteristic syndrome of inflammatory response is serial and complex physiological response such as the enhancement of enzyme activity, the release of inflammatory mediators, fluidal infiltration, cell movement, tissue disruption together with external syndromes such as erythema, edema, pyrexia, ache and so on.
  • NO nitric oxide
  • NOS nitric oxide synthase
  • L-arginine L-arginine to form final products, i.e., NO and citrulline through an intermediate (hydroxyarginine).
  • the substance has small molecular weight and it has been found that it acts on blood vessel system to induce vasodilation, platelet agglutination and adhesion, neuronal transmission, gastrointestinal movement and plays a important roles in controlling metabolic pathway and physiological reaction such as neuronal transmission, blood coagulation, blood pressure regulation and the immunity against cancer cell etc. It is highly toxic because of its free radical structure and is prone to be changed to stabilized final product i.e., NO 3 and NO 2 in the air (Snyder S. H., et al, Scientific American, May pp 28-35, 1992).
  • NOS can be classified with cNOS (constitutive NOS) and iNOS (inducible NOS) according to the dependence to calcium ion or calmodulin; wherein cNOS dependent to calcium ion or calmodulin is present mainly in brain, epithelial cell, neutrophil, stomach mucous cell and; wherein iNOS independent to calcium ion or calmodulin is present mainly in macrophage, hepatic cell, cancer cell etc and is induced by several factors, for example, cytokines such as IL-1beta, IFN-gamma, TNF-alpha, or an endotoxin such as bacterial LPS (Dinerman, J. L., et al, Circ.
  • cytokines such as IL-1beta, IFN-gamma, TNF-alpha
  • endotoxin such as bacterial LPS (Dinerman, J. L., et al, Circ.
  • iNOS expression is closely correlated with COX-2 expression, therefore, formed NO may affects on (C)X-2 expression (Robert C., et al., J. Immunol., 16, pp 1582-1587, 2000; Daniela S., et al., Proc. Nat'l. Acad. Sci. USA, 90 pp 7240-7244, 1993).
  • bamboo belonged to Bambusaceae or Poaceae is distributed in Asian countries including Korea and Japan. There are about 1259 species of bamboo all over the world.
  • the representative ones belonged to Bambusaceae are Phyllostachys bamibusoides SIEB. Et Zucc, Phyllostachys nigra MUNRO, Phyllostachys nigra MUNRO var. henonis STAPF and Phyllostachys pubescens MAZEL ex H. de LEH
  • the representative ones belonged to another Poaceae are Sasa borealis Makino, Sasa coreana Nakai, Sasa japonica Makino, Sasa borealis var. gracilis, Sasa palmata Nakai, Setaria viridis BEAUV and Oryza sativa L.
  • Korean Patent Publication No. 10-2001-69130 discloses on the process for preparing the leave extract from Sasa japonica Makino and the use of the same as a food preservative using its antimicrobial activity
  • U.S. Pat. No. 3418311 discloses the polysaccharide isolated from bamboo having anticancer activity.
  • the inventors of the present invention have intensively carried out several biological experiments i.e., in vitro inhibition test on NO or PLA2 production induced by LPS activated macrophage and an effects on the expression of several gene such as u-PA, eNOS and VEGF known to play an important role in thrombolytic activity, the control of blood flow and the cell growth in blood vessel together with cytotoxicity test, as well as animal model test using LDL receptor defected mouse and normal mouse and finally completed present invention by confirming that the extract and the compound isolated therefrom have the treating and preventing activity on inflammatory or blood circulation diseases.
  • biological experiments i.e., in vitro inhibition test on NO or PLA2 production induced by LPS activated macrophage and an effects on the expression of several gene such as u-PA, eNOS and VEGF known to play an important role in thrombolytic activity, the control of blood flow and the cell growth in blood vessel together with cytotoxicity test, as well as animal model test using LDL receptor defected mouse and normal mouse and finally completed
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising bamboo extract or the compound isolated therefrom as an active ingredient in an effective amount to treat and prevent inflammatory disease caused by the over-production of NO.
  • the present invention provides a pharmaceutical composition comprising bamboo extract or the compound isolated therefrom as an active ingredient in an effective amount to treat and prevent blood circulation disease.
  • the present invention also provides a use of above extract or compound for the preparation of pharmaceutical composition to treat and prevent inflammatory disease and blood circulation disease.
  • the present invention also provides a health care food comprising above extract or compound for the prevention or alleviation of inflammatory disease by inhibiting NO production and blood circulation disease.
  • a pharmaceutical composition comprising the crude extract, polar solvent soluble or non-polar solvent soluble extract of bamboo plant as an active ingredient for the treatment and prevention of cardiovascular disease.
  • It is an object of the present invention to provide a pharmaceutical composition comprising the crude extract, polar solvent soluble or non-polar solvent soluble extract of bamboo plant as an active ingredient for the treatment and prevention of blood circulation disease by inhibiting elastase activity, healing the wound of vascular endothelial cell, activating u-PA expression, inhibiting PAI-1 expression, lowering cholesterol deposit and inhibiting neointima formation.
  • It is an object of the present invention to provide a pharmaceutical composition comprising the crude extract, polar solvent soluble or non-polar solvent soluble extract of bamboo plant as an active ingredient for the treatment and prevention of inflammation by inhibiting NO production and phospholipase A expression.
  • crude extract disclosed herein comprises the extract prepared by extracting plant material with water, lower alcohols such as methanol, ethanol, preferably methanol and the like, or the mixtures thereof.
  • polar solvent soluble extract disclosed herein can be prepared by extracting above crude extract with polar solvent, for example, water, lower alcohol such as methanol, ethanol, preferably butanol and the like, or the mixtures thereof.
  • polar solvent for example, water, lower alcohol such as methanol, ethanol, preferably butanol and the like, or the mixtures thereof.
  • non-polar solvent soluble extract disclosed herein 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.
  • it is another object of the present invention to provide a pharmaceutical composition comprising tricin and p-coumaric acid isolated from bamboo plant extract as an active ingredient for the treatment and prevention of cardiovascular disease.
  • It is an object of the present invention to provide a pharmaceutical composition comprising tricin and p-coumaric acid isolated from bamboo plant extract as an active ingredient for the treatment and prevention of blood circulation disease by inhibiting elastase activity, healing the wound of vascular endothelial cell and increasing VEGF, u-PA and eNOS gene expression.
  • It is another object of the present invention to provide a pharmaceutical composition comprising tricin and p-coumaric acid isolated from bamboo plant extract as an active ingredient for the treatment and prevention of inflammation by inhibiting NO production.
  • the term ‘bamboo plant’ disclosed herein comprises the stem or leaves of bamboo plant belonged to Bambusaceae or Poraceae.
  • Preferable plants belonged to Bambusaceae are Phyllostachys bambusoides SIEB. Et Zucc, Phyllostachys nigra MUNRO, Phyllostachys nigra MUNRO var. henonis STAPF and Phyllostachys pubescens MAZEL ex H. de LEH and more preferable one is Phyllostachys nigra MUNRO var. henonis STAPF.
  • Preferable plants belonged to Poaceae are Sasa borealis Makino, Sasa coreana Nakai, Sasa japonica Makino, Sasa borealis var. gracilis, Sasa palmata Nakai, Setaria viridis BEAUV and Oryza sativa L and more preferable one is Sasa borealis Makino.
  • u-PA urokinase type plasminogen activator gene (fibrinolytic factor)
  • PLA 2 phospholipase A 2 gene
  • VEGF vascular endothelial growth factor gene
  • eNOS endothelial nitrous oxide synthase gene
  • PAI-1 plasminogen activator inhibitor 1 gene.
  • It is an object of the present invention to provide a method of treating and preventing inflammatory disease by inhibiting NO production in a mammal comprising administering to said mammal an effective amount of crude extract, polar solvent soluble or non-polar solvent soluble extract of bamboo extract, together with a pharmaceutically acceptable carrier thereof.
  • It is an object of the present invention to provide a method of treating and preventing blood circulation disease by inhibiting elastase activity and healing the wound of vascular endothelial cell, lowering cholesterol deposit and inhibiting neointima formation in a mammal comprising administering to said mammal an effective amount of crude extract, polar solvent soluble or non-polar solvent soluble extract of bamboo extract, together with a pharmaceutically acceptable carrier thereof.
  • cardiovascular disease comprises various cardiovascular diseases such as hypertension, heart disease, brain stroke, peripheral blood disease, hyperlipemia, arteriosclerosis, stenosis, thrombosis or cardiac infarction etc.
  • inflammatory disease comprises various inflammatory diseases such as atheriosclerosis, arthritis, gastritis, colitis, nephritis, hepatitis, cancer or various degenerative diseases.
  • the pharmaceutical composition of the present invention can contain about 0.1 ⁇ 70% by weight of the above extract or compound based on the total weight of the composition.
  • the health care food of the present invention comprises the above extract or compound as 0.01 to 80%, preferably 1 to 60% by weight based on the total weight of the composition.
  • Above health care food can be contained in health care food, health beverage etc, and may be used as powder, granule, tablet, chewing tablet, capsule, beverage etc.
  • An inventive extract and compound isolated from bamboo plant may be prepared in accordance with the following preferred embodiment.
  • An inventive extract of bamboo plant can be prepared in detail by following procedures.
  • the inventive crude extract of Phyllostachys nigra MUNRO var. henonis STAPF or Sasa borealis Makino can be prepared by follows; Phyllostachys nigra MUNRO var. henonis STAPF or Sasa borealis Makino 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 1 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.,
  • 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 crude extract, polar solvent soluble or non-polar solvent soluble extract of Phyllostachys nigra MUNRO var. henonis STAPF or Sasa borealis Makino prepared by above preparation method or the treatment and prevention of inflammation by inhibiting NO production as active ingredients.
  • the inventive composition for treating and preventing inflammation by inhibiting NO production and for improving blood circulation may comprises above extracts as 0.1 ⁇ 70% by weight based on the total weight of the composition.
  • the 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 or 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.
  • 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).
  • oral dosage form prowder, tablet, capsule, soft capsule, aqueous medicine, syrup, elixirs pill, powder, sachet, granule
  • topical preparation cream, ointment, lotion, gel, balm, patch, paste, spray solution, aerosol and the like
  • 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 compound varies depending on the condition and the weight of the subject, severity, drug form, route and a 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 70% by weight, preferably 0.5 to 50% 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.
  • health care food disclosed herein comprises dietary supplement, nutraceuticals, food or food additives.
  • the present invention provide a composition of the health care food beverage for the prevention or improvement of inflammation or blood circulation 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 allergic disease and non-allergic inflammation disease by adding to child and infant food, such as modified milk powder, modified milk powder for a growth period, modified food for a 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 care food composition and 1 to 30 g, preferably 3 to 10 g on the ratio of 100 ml 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, cyclodextrin; 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 ml 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 ibod comprising aforementioned 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, alpha-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, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.
  • the above extract of bamboo plant may be 20 to 90% high concentrated liquid, power, or granule type.
  • the above extract of bamboo plantcan 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.
  • FIGS. 1 and 2 represent the HPLC analysis data of tricin and p-coumaric acid, FIG. 1 is for standard group, FIG. 2 is for inventive bamboo extract;
  • FIGS. 3 and 4 show the effects of various concentrations of bamboo crude extract and fractions isolated therefrom on NO inhibition, FIG. 3 is for 50 ?g/ml treated group,
  • FIG. 4 is for 100 ?g/ml treated group, wherein
  • A crude extract
  • B n-hexane soluble extract
  • E ethyl-acetate soluble extract
  • F water-soluble extract
  • FIG. 5 shows the cellular toxicity of bamboo extract, tricin and p-coumaric acid in the HUVEC, wherein the numbers described above the graph bars denote the concentration of treated sample (?g/ml );
  • FIG. 6 shows the cell proliferation effect of bamboo extract in the HUVEC
  • FIG. 7 to 9 show the effect of bamboo extract, tricin and p-coumaric acid in the HUVEC with complete media on mRNA expression
  • FIG. 7 is for VEGF expression
  • FIG. 8 is for u-PA expression
  • FIG. 9 is for eNOS expression, wherein the numbers described above the graph bars denote the concentration of treated sample (?g/ml)
  • FIG. 10 to 12 show the wound healing effect of the inventive bamboo extract through in vitro wound healing assay using HUVEC
  • FIG. 10 is for control group
  • FIG. 14 is for 10 ?g/ml bamboo extract-treated group
  • FIG. 12 is for 50 ?g/ml of bamboo extract-treated group;
  • FIG. 13 to 15 show the blood vessel formation of the inventive bamboo extract through in vitro tube formation assay using HUVEC, FIG. 13 is for control group, FIG. 14 is for 10 ?g/ml of bamboo extract-treated group and FIG. 15 is for 100 ?g/ml of bamboo extract-treated group;
  • FIG. 16 shows the change of body weights in the high cholesterol diet induced atherosclerosis mice with or without the treatment of bamboo extract for 16-weeks (50 and 100 mg/g) and 20-weeks (500 mg/kg);
  • FIGS. 17 and 18 show the morphometry of Oil red O stained aortic valve lesion areas by computer-associated mage analysis in the high cholesterol diet induced atherosclerosis mice with or without the treatment of bamboo extract for 16-weeks (50 and 100 mg/kg) and 20-weeks (500 mg/kg),
  • FIG. 17 represents Oil red O stained photographs of frozen sections of aortic valve lesion, wherein the left panel is for control group, the middle panel is for positive control group treated with Lovastatin and right panel is for bamboo extract-treated group, and FIG. 18 represents morphometric results of aortic valve lesion areas by computer-associated image analysis.
  • Example 2 50 g of crude extract of Phyllostachys nigra prepared in Example 1 was suspended in 1 liter of distilled water and the suspension was mixed with 1 liter of n-hexane vigorously to divide into n-hexane soluble fraction and water-soluble fraction. n-hexane soluble fraction was collected and the residual solution was subjected to the n-hexane extraction again. Above-described procedure was repeated 3 times.
  • n-hexane soluble fraction was evaporated in vacuo to give 9.1 g of n-hexane soluble extract of Phyllostachys nigra.
  • Dichloromethanesoluble fraction was evaporated in vacuo to give 4.6 g of dichloromethane soluble extract of Phyllostachys nigra.
  • Example 2-2 Water-soluble fraction of Phyllostachys nigra in Example 2-2 was mixed with equivalent volume of ethylacetate vigorously to divide into ethylacetate soluble fraction and water-soluble fraction. Ethylacetate soluble fraction was collected and the residual solution was subjected to the ethylacetate extraction again. Above-described procedure was repeated 3 times.
  • n-butanol soluble fraction and water-soluble fraction were respectively evaporated in vacuo to give 7.1 g of n-butanol soluble extract and 25.1 g of water-soluble extract of Phyllostachys nigra.
  • each polar solvent and non-polar solvent soluble extract was prepared according to the identical method disclosed in the above Example 2.
  • TABLE 2 Amount Crude extract 57 g n-hexane soluble extract 9.5 g Dichloromethane soluble extract 4.1 g Ethyl acetate soluble extract 4.8 g n-butanol soluble extract 27.9 g Water-soluble extract 27.9 g
  • Example 1 100 g of the crude extract obtained in Example 1 was suspended in 1000 ml of distilled water. 1000 ml of chloroform was added thereto in separatory funnel and the mixture was shaken vigorously to divide into chloroform soluble layer and water soluble layer. Chloroform soluble fraction was collected and the residual solution was subjected to the chloroform extraction again.
  • water-soluble layer was also obtained to use as a sample in the following experiments.
  • the 7 th fraction was subjected to preparation HPLC to obtain 4.2 mg of phenyl propanoid compound and the isolated phenylpropanoid compound was identified as p-coumaric acid derivatives by the result of 1 H-NMR spectrum by NMR spectroscopy ( 1 H: 300 MHz, DRX 300, Bruker, Germany) shown as below and the retention time of HPLC analysis compared with the data of p-coumaric acid standard purchased from Sigma Company (See FIGS. 1 and 2 ).
  • Murine macrophage cell line RAW 264.7 cell (ATCC, Rockville, Md., USA) were grown in DMEM (Gibco BRL Co., Ltd., USA), supplemented with 2.0 mM Larginine, 100 ?g /ml penicillin-streptomycin and 10% fetal bovine serum at 37° C. in 5% CO 2 and 95% air condition in humidified incubator.
  • HUVEC Human umbilical vein endothelial cell
  • MTT Co. gelatin coating flask
  • EGM-2 media Clonetics (C.) supplemented with 20% FBS, 100 ⁇ antibiotics and 200 ⁇ ECGF and the cells passaged 3 to 5 times were used in the following experiment.
  • arteriosclerosis model mice were used in the experiment.
  • mice under the artificial lamp for 12 hours, and less than five mice were bred in each mouse cage providing with free access to water (disinfected distilled water) and normal fatty feed(Harlan 2018S, Indianapolis USA).
  • high fat feed Harlan TD88051, Artherosclerotic diet, overall fat content is about 15.8%; cholesterol level of 1.25%, and sodium cholate at 0.5%, about 4 kcal/g, and 35% of kcal from fat. About half the fat come from added cocoa butter, and half from the chow) were provided.
  • NO nitric oxide
  • the inventive bamboo extract was treated with elastase enzyme, which degrades elastin protein in charge of maintaining elasticity and strength of blood vessel.
  • Example 1-5 Each bamboo extract or fraction prepared in Example 1-5 was diluted to 20, 2 and 0.2 mg/ml and aliquoted by 50 ?1 into each 96 well plate.
  • Commercial elastase (Molecula probe Co.) was added thereto at the concentration of 0.5 U/ml and elastin protein was also added at the concentration of 50 ?g/ml.
  • absorbance was detected using ELISA reader.
  • HUVEC was grown confluently on 0.2% gelatin-coated 12-well plate and then scratched by cell scraper to make original wound edge.
  • the cells were treated with 10 ?g/ ml or 50 ?g/ml of bamboo extract and cultured in 5% CO 2 incubator. The translocation of the cell was deserved by the pictures.
  • the cells were treated with 10 ?g/ml or 50 ?g/ml of bamboo extract and cultured in 5% CO 2 incubator.
  • the morphological change of the HUVEC was observed under microscope at regular interval and taken the picture.
  • the extract or compound of the present invention was treated to the cell and the RNA extracted therefrom was used in the RT-PCR to evaluate the quantitative gene expression.
  • RT-PCR was performed according to the RT reaction (25° C. 10 min, 48° C. 30 min, 95° C. 5 min, 4° C. 10 min; 1 cycle) and subsequent PCR (50° C. 2 min, 95° C. 10 min, 95° C. 15 sec, 60° C. 1 min, 40 cycles) method well known in the art.
  • 18S ribosomal RNA was used as an internal control.
  • OD260/OD280 value of extracted RNA determined by Spectrophotometer was more than 1.7 and the purity of RNA was confirmed by Denaturing agarose gel electrophoresis.
  • RT-PCR was performed according to the RT reaction (10 min at 25° C., 30 min at 48° C., 5 min at 95° C., 10 min at 4° C.; 1 cycle) and subsequent PCR (2 min at 50° C., 10 min at 95° C., 15 sec at 95° C., 1 min at 60° C., 40 cycles) method well known in the art.
  • 18S ribosomal RNA was used as an internal control.
  • RNA extraction and RT-PCR were performed according to the method above described in Experimental Example 4-1.
  • RNA was isolated from HUVEC cell by Rneasy mini kit (cat# 74103, Qiagen Co.) according to the manufacturer's instruction. And RT-PCT reaction was performed by using quantitative PCR method (SDS 7700, Applied biosystems Co., U.S.A.).
  • tricin compound treatment to HUVEC had increased the expressions of VEGF (vascular endothelial growth factor), u-PA gene and the expression of eNOS (endothelial nitrous oxide synthase), which affects the vascular expansion in atrophy.
  • VEGF vascular endothelial growth factor
  • u-PA vascular endothelial growth factor
  • eNOS endothelial nitrous oxide synthase
  • mice were anesthetized with 0.12% of avertin and exsanguinations was performed from infraorbitalis plexus venosus with heparin treated capillary. And then blood plasma was isolated by centrifugation at the speed of 11,000 g for 10 minutes and left alone at ⁇ 70° C. before use.
  • the value of blood lipid was determined by three categories i.e., TC (Total cholesterol), HDL-C (High-density lipoprotein cholesterol) and TG (Triglycerides) at KRIBB in Korea.
  • mice were anesthetized with 0.12% of avertin and exsanguinations was performed from infraorbitalis plexus venosus with heparin treated capillary. And then blood plasma was isolated by centrifugation at the speed of 11,000 g for 10 minutes and left alone at ⁇ 70° C. before use.
  • the value of blood lipid was determined by three categories i.e., TC (Total cholesterol), HDL-C (High-density lipoprotein cholesterol) and TG (Triglycerides) at KRIBB in Korea.
  • the exsanguinated heart was fixed with 4% paraformaldehyde dissolved in 0.1M phosphate buffer (pH 7.4) and delivered removing remaining blood and fixing with 10% neutral formalin. And then it is embedded with OCT compound, sliced into 0.6 ⁇ m of thickness, stained with oil red O and count-stained with Harris hematoxylin to observe the lesion.
  • lesion area was calculated by staining the lesion formed at the position between 3 rd cervical blood and aortic valve and photocopying and then the lesion area was calculated by using computer-assisted morphometry (TDI microscope Image Analyzer, USA) comparing with control group.
  • sample treatment group inhibit the formation of arteriosclerosis by about 17% compared with control group while lovastatin used as a positive control inhibit by about 47% and prevent the formation of neointima (See FIGS. 17 and 18 ).
  • HUVEC 200 ?1 of HUVEC (2 ⁇ 10 5 cells/ml) on flat bottom 96-well microtiter plates (Nunc, Sweden) were treated with tricin prepared in various concentrations and cultured at 37° C. for 24 hours.
  • MTT solution (1 mg/ml) was added to each well and incubated at 37° C. for 4 hours. And then supernatant was removed.
  • the inventive tricin compound showed the 51% of strong cellular toxicity at 5 ?g/ml, however, bamboo extract or p-coumaric acid was no cellular toxicity (See FIG. 5 ).
  • the cell proliferation assay of bamboo extract was performed using Cell Proliferation ELISA BrdU colorimetric kit (Roche). HUVEC were seed 5 ⁇ 10 3 cells/well in 95 well plate. Triplicate plate of cells were measured using ELISA Reader.
  • bamboo extract enhanced strong cellular proliferation by dose dependent manner (See FIG. 6 )
  • mice mean body weight 25 ⁇ 5 g
  • Sprague-Dawley rats 235 ⁇ 10 g, Jung-Ang Lab Animal Inc.
  • test sample or solvents 0.2 ml, i.p.
  • mice and rats were treated with the extract of the Example 1.
  • mice or rats was administrated intraperitoneally with 25 mg/kg, 250 mg/kg, 500 mg/kg and 725 mg/kg of test sample or solvents (0.2 ml, i.p.), respectively and observed for 24 hours.
  • Powder preparation was prepared by mixing above components and filling sealed package. Preparation of tablet Dried powder of Example 1 50 mg Corn Starch 100 mg Lactose 100 mg Magnesium Stearate 2 mg
  • Tablet preparation was prepared by mixing above components and entabletting. Preparation of capsule Dried powder of Example 1 50 mg Corn starch 100 mg Lactose 100 mg Magnesium Stearate 2 mg
  • Tablet preparation was prepared by mixing above components and filling gelatin capsule by conventional gelatin preparation method. Preparation of injection Dried powder of Example 1 50 mg Distilled water for injection optimum amount PH controller optimum amount
  • Injection preparation was prepared by dissolving active component, controlling pH to about 7.5 and then filling all the components in 2 ml ample and sterilizing by conventional injection preparation method.
  • Preparation of liquid Dried powder of Example 1 0.1 ⁇ 80 g Sugar 5 ⁇ 10 g Citric acid 0.05 ⁇ 0.3% Caramel 0.005 ⁇ 0.02% Vitamin C 0.1 ⁇ 1% Distilled water 79 ⁇ 94% CO 2 gas 0.5 ⁇ 0.82%
  • Liquid preparation was prepared by dissolving active component, filling all the components and sterilizing by conventional liquid preparation method.
  • Preparation of health care food Extract of Example 1 1000 mg Vitamin mixture optimum amount ? Vitamin A acetate 70 mg ? Vitamin E 1.0 mg ? Vitamin B 1 0.13 mg ? Vitamin B 2 0.15 mg ? Vitamin B 6 0.5 mg ? Vitamin B 12 0.2 mg ? Vitamin C 10 mg ? Biotin 10 mg ? Amide nicotinic acid 1.7 mg ? Folic acid 50 mg ? Calcium pantothenic acid 0.5 mg Mineral mixture optimum amount ? Ferrous sulfate 1.75 mg ? Zinc oxide 0.82 mg ? Magnesium carbonate 25.3 mg ? Monopotassium phosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg
  • 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 1000 ml ample and sterilizing by conventional health beverage preparation method.
  • the bamboo plant extract and the tricin compound therefrom have potent anti-inflammatory activity by inhibiting NO production and PLA expression, blood circulation-improving activity by inhibiting elastase activity and healing the wound of vascular endothelial cell, activating u-PA expression and inhibiting PAI-1 expression, lowering cholesterol deposit and inhibiting neointima formation, therefore, it can be used as a therapeutic, health care food for treating and preventing inflammatory or blood circulation diseases.

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KR1020030019229A KR100417243B1 (ko) 2003-03-27 2003-03-27 항염증 활성을 지닌 분죽 추출물을 함유하는 염증성 질환의 예방 및 치료용 약학조성물
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US20080233242A1 (en) * 2003-10-08 2008-09-25 Ying Zhang Antioxidant of Bamboo Leaves and Its Uses
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US10813968B2 (en) 2012-12-24 2020-10-27 Provexis Natural Products Limited Method of treating a human suffering from post exercise muscle soreness
US10905733B2 (en) 2016-11-02 2021-02-02 Provexis Natural Products Limited Water soluble tomato extract protects against adverse effects of air pollution
US10918690B2 (en) * 2018-09-06 2021-02-16 Louise Wilkie Apparatus and method for processing organic bamboo leaf extract products
US20200078433A1 (en) * 2018-09-06 2020-03-12 Louise Wilkie Apparatus and method for processing organic bamboo leaf extract products
US20210161989A1 (en) * 2019-04-06 2021-06-03 Louise Wilkie Apparatus and method for processing organic bamboo leaf extract products
US11786573B2 (en) * 2019-04-06 2023-10-17 Louise Wilkie Apparatus and method for processing organic bamboo leaf extract products

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US20110117224A1 (en) 2011-05-19
AU2004235702A1 (en) 2004-11-18
JP2006515007A (ja) 2006-05-18
BRPI0408814A (pt) 2006-04-04
CA2520207C (fr) 2010-08-17
US7897182B2 (en) 2011-03-01
JP4684893B2 (ja) 2011-05-18
EP1633379A4 (fr) 2009-09-02
EP2213294A1 (fr) 2010-08-04
CA2520207A1 (fr) 2004-11-18
EP1633379A1 (fr) 2006-03-15
AU2004235702B2 (en) 2008-07-31
WO2004098624A1 (fr) 2004-11-18
US20110280975A1 (en) 2011-11-17

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