CN1275906A - Plaque-inhibiting oral compositions - Google Patents
Plaque-inhibiting oral compositions Download PDFInfo
- Publication number
- CN1275906A CN1275906A CN98810170A CN98810170A CN1275906A CN 1275906 A CN1275906 A CN 1275906A CN 98810170 A CN98810170 A CN 98810170A CN 98810170 A CN98810170 A CN 98810170A CN 1275906 A CN1275906 A CN 1275906A
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- China
- Prior art keywords
- bacterial strain
- plaque
- oral cavity
- enzyme
- bacillus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
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- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 description 1
Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- 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
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
- A61Q11/02—Preparations for deodorising, bleaching or disinfecting dentures
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Cosmetics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to oral compositions comprising plaque-inhibiting or plaque-removing enzymes, in particular at least one starch-hydrolysing enzyme, e.g. an alpha-amylase or a debranching enzyme such as a pullulanase, and/or at least one starch-modifying enzyme, e.g. a transglucosidase or a CGTase, and to a method for inhibiting plaque formation or removing plaque using such oral compositions.
Description
Technical field that the present invention belongs to
The present invention relates to include the composition for oral cavity of plaque inhibitory enzyme or plaque removal enzyme, also relate to the method for utilizing these composition for oral cavity plaque-inhibitings to form or remove plaque.
Background of invention
Dental plaque is an antibacterial, epithelial cell, leukocyte, the mixture that macrophage and other oral cavity exudate form on unclean dental surface.Bacteriogenic hyperbranched polysaccharide and the coherent matrix that forms the continuous propagation of plaque from intraoral microorganism jointly.If do not handle, the formation of dental plaque will finally cause dental caries, gums inflammation, periodontal, and final loss of tooth.When plaque continuously accumulation the time, hard white or faint yellow deposit as the rock appears.These deposits are called the calcification plaque, calculus or tartar, and by plaque and mineral, particularly calcium forms in saliva.
The oral cavity polysaccharide is by the sucrose of for example introducing the oral cavity as food or beverage composition, by growing in the livings dental caries microorganism in the oral cavity such as the effect generation of mutan streptococcus (Streptococcus mutans) or Streptococcus sanguis (Streptococcus sanguis).These oral cavity polysaccharide comprise that water-soluble glucan (mainly contains α-1,6 glycosidic bonds), and key component is a kind of is called the water-insoluble extracellular polysaccharide of " mutan ", and it is by having α-1, the main chain of 3-glycosidic bond and have a α-1, the side chain of 6-glycosidic bond is formed.Mutan is attached on the hydroxyapatite (constituting the hard porous outer layer of tooth), is attached on the receptor protein on living dental caries bacterial cell surface again, and said living dental caries bacterial adhesion is on dental surface.
People have carried out a large amount of trial work, thereby form environment by eliminating plaque or changing plaque by the composition that changes plaque, to stop the formation of plaque.Aspect removal or plaque-inhibiting, many toothpaste and other oral cavity health protection have obtained success in various degree with compositions.Yet the purpose of plaque-inhibiting does not also realize effectively, various oral cavity health compositionss of demand still, by daily use just effectively plaque-inhibiting formation with can form the property the followed odontopathy that causes by plaque.
WO 97/06775 discloses the composition for oral cavity that is used for bleaching teeth, comprises oxidoreductase, and optionally glucanase and/or mutanase, but does not mention or point out the plaque-inhibiting of this compositions or remove the plaque effect.
In order to prevent dental caries, the formation of dental plaque and tartar, suggestion are used in oral cavity health and are added glucanase and/or mutanase and/or other enzyme in compositions and the product.
JP patent disclosure 8012544 (Lion) has been described glucanase, the prevention plaque effect of mutanase and triclosan and/or biosol.
United States Patent (USP) 4,353,891 (Guggenheim etc.) mention the mutan enzymatic degradation of using from Trichodermaharzianum CBS 243.71 and remove dental plaque by cultivating the synthetic mutan of mutan strains of streptococcus CBS 350.71 (can differentiate and be OMZ 176).According to this article record, the key component in the dental plaque is to have α-1, the water-insoluble polysaccharide of 3-glycosidic bond, and also the polysaccharide material of this being called " mutan " is not subjected to the effect of glucanase.
Guggenheim etc. (1972), dental caries research 6, p.289-297, the degree that has disclosed the formation of rat dental plaque is not to be subjected to using simultaneously glucanase and α-1, the appreciable impact of 3-glucanase (mutanase).
Hare etc. (1978), p.245-264 carbohydrate compound research 66, is found with bacterial dextran enzyme and bacterial alpha-1 from Cladosporium resinae, the 3-glucanase can obtain synergism during in conjunction with hydrolysis and solubilising oral cavity glucosan.
United States Patent (USP) 4,438,093 (Osaka microbial diseases WARF) described and included glucanase and α-1, the composition for oral cavity of 3-glucanase (mutanase), the amount of two kinds of enzymes is 0.5 to 100 enzyme unit in the said oral cavity composition of every gram, and the enzyme unit ratio is 1: 2 to 2: 1.Said glucanase is from the antibacterial in the Corynebacterium, said α-1, and the 3-glucanase is from the antibacterial in the Rhodopseudomonas.
GB 2,206, and 585 (dental chemistry company limiteies) have been described a kind of tooth cleaning agent, contain hydroxyapatite as abrasivus, laevanase, and glucanase and mutanase are fixed on the hydroxyapatite.
United States Patent (USP) 5,145,665 (Henkel) disclose a kind of compositions that is used for oral cavity and dental care, and it contains glucanase and/or α-1, and the 3-glucanase is used for the polysaccharide in enzymolysis oral cavity.
FR 2,651, and 433 (DANA) relate to the dentifrice product, and it contains: act on the glucanase of nearest plaque, act on old and mutanase insoluble plaque and the mixture with other enzyme of bactericidal action.
United States Patent (USP) 5,320,830 (Proctor ﹠amp; Gamble) described the dentifrice composition that is used to reduce plaque and gingivitis, it comprises a) surfactant), b) enzyme, C) chelating agen, d) fluoride source, e) Silicon stone abrasive material and f) carrier.Enzyme is an endoglucanase, papain, glucanase and/or mutanase.
Now being surprised to find the oral cavity health that includes one or more amylolytic enzymes or starch modification enzyme is effective with compositions for the formation of inhibition/prevention dental plaque and/or for removing plaque.
Summary of the invention
The object of the present invention is to provide and effectively to suppress/stop the composition for oral cavity that plaque formed and removed plaque, also provide a kind of plaque-inhibiting to form or remove the method for plaque.
A first aspect of the present invention relates to a kind of oral cavity health compositions, at least a amylolytic enzyme and/or at least a starch modification enzyme that this oral cavity health comprises plaque-inhibiting and/or removes the plaque effective dose with compositions.
Second aspect, the present invention relates to a kind of method that plaque-inhibiting formed or removed plaque that is used for, this method comprises makes tooth and/or chewing gum contact a period of time to reach the effect of plaque-inhibiting or removal plaque with at least a amylolytic enzyme that includes plaque-inhibiting and/or removal plaque effective dose and/or at least a starch modification enzyme's oral cavity health with compositions.
The third aspect, the present invention relates to one or more amylolytic enzymes and/or one or more starch modification enzyme be used in preparation suppressing/stop plaque formation and/remove the application of the compositions of plaque.Detailed description of the present invention
Term in the application's context " amylolytic enzyme " refers to any enzyme, and as α-Dian Fenmei (E.C.3.2.1.1), its function is the key in the hydrolyzed starch.
Term in the application's context " starch modification enzyme " refers to the enzyme cohort in E.C.2.4.1., especially comprises transglucosidase (E.C. 2.4.1.18) and CGT enzyme (E.C.2.4.1.19).
The inventor has been found that by utilizing at least a amylolytic enzyme or at least a starch modification enzyme, can suppress/stop or remove dental plaque.Former people also do not predict this kind effect of these enzymes.
A first aspect of the present invention relates to a kind of oral cavity health compositions, and said composition comprises at least a amylolytic enzyme and/or at least a starch modification enzyme who suppresses and/or remove the plaque effective dose.
The starch modification enzyme is CGT enzyme (E.C.2.4.1.19) or transglucosidase (2.4.1.18) in an embodiment of the present invention.
When the starch modification enzyme is the CGT enzyme, it may be derived by following bacterial strain: the bacterial strain of Bacillusautolyticus, the bacterial strain of bacillus cereus, the bacterial strain of Bacillus circulans, Bacillus circulans is had a liking for the bacterial strain of alkali mutation, solidify the bacterial strain of bacillus cereus, the bacterial strain of strong bacillus cereus, the bacterial strain of Bacillus halophilus, the bacterial strain of bacillus macerans, the bacterial strain of bacillus megaterium, the bacterial strain of Bacillus ohbensis, the bacterial strain of bacstearothermophilus, the bacterial strain of bacillus subtilis, the bacterial strain of Klebsiella pneumonia, hot anaerobic bacillus(cillus anaerobicus) belongs to the bacterial strain of strain, the dwell bacterial strain of hot anaerobic bacillus(cillus anaerobicus) of the bacterial strain of the hot anaerobic bacillus(cillus anaerobicus) of producing and ethanol, fin, the bacterial strain of pyrolysis clostridium amylobacter, the bacterial strain of the bacterial strain of clostridium thermosaccharolyticum or the hot anaerobic bacillus(cillus anaerobicus) of delivery in hot weather sulfur.
When the starch modification enzyme was transglucosidase, it may be derived from aspergillus niger, the product that for example Japanese Amamo pharmaceutical companies is sold.
Oral cavity health comprises a kind of amylolytic enzyme with compositions in another embodiment of the present invention.
This kind of enzyme is typically α-Dian Fenmei, as bacterial, as BAN
TMPerhaps Maltogenase
TM(all can be obtained by Novo Nordisk) is perhaps from the deutero-α-Dian Fenmei of bacillus subtilis; From the deutero-α-Dian Fenmei of bacillus amyloliquefaciens; From the deutero-α-Dian Fenmei of bacstearothermophilus; From the deutero-α-Dian Fenmei of aspergillus oryzae; Perhaps from the deutero-α-Dian Fenmei of non-pathogenic microorganism.
α-Dian Fenmei can be a fungal alpha-amylase also, as Fungamyl
TM, can obtain by Novo Nordisk.
Amylolytic enzyme may be debranching enzyme, particularly amylopectase (E.C.3.2.1.41) in another embodiment of the present invention, as Promozyme
TM
In an embodiment preferred, oral cavity health comprises at least a modification enzyme of starch as defined above, particularly CGT enzyme, and mutanase and/or glucanase with compositions.
In another embodiment preferred, oral cavity health of the present invention comprises at least a amylolytic enzyme as defined above, particularly bacterial with compositions, and mutanase and/or glucanase.
Mutanase can be derived from the bacterial strain of trichoderma strain (Trichoderma sp.), especially T.harzianum, particularly T.harzianum CBS 243.71 (can be obtained by Novo Nordisk).
Glucanase can be derived from the bacterial strain of paecilomyces strain (Paecilomyces sp.), especially Paecilomyces lilacinus (can be obtained by Novo Nordisk).
A second aspect of the present invention relates to a kind of oral health products that contains oral cavity health of the present invention with compositions.
" oral health products " of the present invention is defined as and a kind ofly can be used for keeping and/or improve human and animal's the oral hygiene and/or the product of prevention or treatment odontopathy.
The example of such oral health products comprises toothpaste, the tooth frost, tooth gel or dentifrice, the tooth irrigation, collutory, tooth cleaning agent, brush teeth preceding or brush teeth after rinsing preparation, chewing gum and lozenge.Oral health products also can be the form of dental floss or toothpick.
Toothpaste and tooth gel typically comprise polishing material, foaming agent, flavouring agent, wetting agent, binding agent, thickening agent, sweeting agent and water.As used herein, term " toothpaste " means the preparation of pastel and two kinds of forms of gel.
Collutory typically comprises water/alcoholic solution, flavouring agent, wetting agent, sweeting agent, foaming agent and coloring agent.
Can prepare with methods known in the art according to chewing gum of the present invention, by one or more enzymes being spiked in the conventional chewing gum base, be substrate for example with the additional synthetic or natural polymer of tunny gum and/or one or more, and contain for example natural and/or synthetical sweeting agent as required, flavouring agent or the like.When enzyme being added in the gummy substrate, the temperature of gummy substrate should be too not high, for example, preferably is not higher than about 60 ℃, more preferably is not higher than about 50 ℃.
According to the present invention, the polishing material that is used for oral health products is aluminium oxide and hydrate thereof, as α-gibbsite, magnesium trisilicate, magnesium carbonate, sodium bicarbonate (sodium bicarbonate), Kaolin, aluminosilicate is as the aluminium silicate and the aluminium silicate of calcination, calcium carbonate, Zirconium orthosilicate., and powdered plastic are as polrvinyl chloride, nylon, polymethyl methacrylate, polystyrene, phenol-formaldehyde resin, melamine formaldehyde resin, urea-formaldehyde resins, epoxy resin, powder polyethylene, silica xerogel, hydrogel and aeroge and analog thereof.What be fit to do grinding agent also has calcium pyrophosphate, non-water-soluble alkaline metaphosphate, dicalcium phosphate and/or its dihydrate, orthophosphoric acid dicalcium, tricalcium phosphate, graininess hydroxyapatite and an analog thereof.Also may use the mixture of these materials.
The character that depends on oral health products, the weight content of abrasive product can from 0 to 70%, and preferably from 1% to 70%.For toothpaste, grinding-material accounts for the weight content of final tooth paste product typically in 10% to 70% scope.
Using wetting agent is in order to prevent moisture loss, and for example moisture loses from toothpaste.According to the present invention, the suitable wetting agent that is used for oral health products comprises following compounds and composition thereof: glycerol, polyhydric alcohol, sorbitol, Polyethylene Glycol (PEG), propylene glycol, 1, ammediol, 1,4-butanediol, the polysaccharide of hydrogenation partial hydrolysis and analog thereof.The weight content of wetting agent in toothpaste is generally 0% to 80%, and preferably from 5 to 70%.
Suitable thickening and binding agent help the stable of dentifrice product, these examples of substances have silicon dioxide, starch, Tragacanth, xanthan gum, the Ai Lantai extract, alginate, pectin, cellulose derivative, as hydroxyethyl-cellulose, sodium carboxymethyl cellulose and hydroxyl-propyl cellulose, polyacrylic acid and salt thereof and polyethylene-ketopyrrolidine.Thickening agent is at toothpaste, and the weight content in frost and the gel is 0.1 to 20%, and the weight content of binding agent in final products is 0.01 to 10%.
Soap, anionic, cationic, non-ionic, surfactant amphoteric and/or hybrid ion can be used as foaming agent.These materials are the weight content from 0 to 15% in the product in the end, and preferably from 0.1 to 13%, more preferably from 0.25 to 10%.
Surfactant only is suitable in the context of the present invention to a certain extent, and promptly they do not have any adverse effect to the activity of enzyme.
The example of surfactant comprises fatty alcohol sulfate, sulfonation monoglyceride or have the fatty acid of 10 to 20 carbon atoms, fatty acid-albumin condensation product, the fatty acid ester salt of fatty acid acyl amine salt and taurine and/or isethionic acid.
Suitable sweeting agent comprises that glucide and/or other are suitable for the sweeting agent of oral health products.
Flavouring agent, as Mentha viridis L and lavender, its weight content in final products is lower usually, for example from 0.01% to about 5%, particularly from 0.1% to 5%.
Common water with a certain amount of adding, but for example make toothpaste become liquid form, and the weight content that promptly accounts for final products is 40% to 70%.
In addition, also can comprise water-soluble antimicrobial, as glucosulfone pickling BITAI, hexetidine, alexidine, the source thing of quaternary ammonium antibacterial compounds and water miscible some metal ion, metal ion such as zinc, copper, silver and stannous ion (for example zinc, copper and stannous chloride and silver nitrate).
According to the present invention, also can consider except that theme-enzyme of the application, to add other antilithic, anti-plaque agent can be used as the chemical compound that fluoride is originated, dyestuff/coloring agent, antiseptic, vitamin, pH-regulator, anti-caries agent, desensitizer or the like.
The toothpaste of being produced by composition for oral cavity of the present invention can comprise, for example following ingredients (% by weight in final dentifrice composition): grinding-material 10-70% wetting agent 0-80% thickener 0.1-20% adhesive 0.01-10% sweetener 0.1-5% foaming agent 0-15% starch degrading enzyme and/or other enzyme of starch modification enzyme 0.0001-20% 0-20% peroxide 0-1%
The collutory of being produced with composition by oral health of the present invention can comprise; Following ingredients (% by weight in final dentifrice composition) for example: 0-20% wetting agent 0-2% surfactant 0-5% starch degrading enzyme and/or other composition of other enzyme of starch modification enzyme 0-5% 0-20% ethanol 0-2% (flavouring agent for example, sweetener
Or other active component, as fluoride) 0-70% water
Mouthwash agent composition can be with suitable buffer agent, and for example the pH scope is in sodium citrate or the phosphate-buffered of 6-7.5.
The form of collutory can be undiluted attitude (i.e. dilution before use) or dilute attitude (stand-by).
Oral cavity health of the present invention can be made of method common in this area of compositions and product.
The invention further relates to aforesaid one or more amylolytic enzymes and/or one or more starch modification enzyme and be used for suppressing/stoping the application that plaque formed and/or removed the compositions of plaque in preparation.
But solid-state oral cavity health product such as toothpaste to flowable state are typically by using toothbrush or its analog and tooth and/or natural gum to contact.Under liquid oral product situation for health care, as collutory, product for health care typically occurs in the process of gargling with contacting of tooth.
Oral health products of the present invention is changed, as the character of compositions and experimenter's demand because of following factors is different with the time that obtains desired plaque-inhibiting or removal plaque effect with tooth and/or natural gum contact.Yet oral health products contacts about 30 seconds with tooth and/or natural gum under normal circumstances can obtain desired effects by 15 minutes, for example, compositions was contacted in about 1-3 minute with tooth by brushing teeth or gargle sometime.This preferably carries out with certain rule, for example, and one day 1-3 time.
After using up, generally to from the oral cavity, remove the oral cavity health product, for example tell it, and gargle with a kind of liquid such as tap water subsequently.
Though do not wish to be subjected to the constraint of any particular theory, but believe that the surprising plaque-inhibiting effect that the present invention finds may be because the following fact, promptly before the enzyme of microorganisms was near sucrose, amylolytic enzyme and/or starch modification enzyme and sucrose reacted and have produced a kind of coupling sugar.No matter what the mechanism of reaction is, uses the observed plaque-inhibiting effect of being discussed of enzyme astonishing, and believe that nobody described or pointed out related content before this.
In following non-limiting example, the present invention will obtain further instruction.Material and method material
Glucanase results from Paecilomyces lilacinus (providing A/S by Novo Nordisk).
Mutanase results from Trichoderma harzianum CBS 243.71 (providing A/S by NovoNordisk).
Produce maltase: Novamyl
TM(antibacterial Fructus Hordei Germinatus α-Dian Fenmei) provides A/S by Novo Nordisk.
Fungamyl
TM(fungal alpha-amylase) can be obtained by Novo Nordisk A/S.
Transglucosidase results from aspergillus niger (can be obtained by the Amona pharmaceutical companies).
BAN
TM(bacterial) can be obtained by Novo Nordisk A/S.
The CGT enzyme results from hot antibacterial and belongs to strain, can be obtained by Novo Nordisk A/S.
Promozyme
TM(antibacterial amylopectase) can be obtained by Novo Nordisk A/S.Microorganism
Can differentiate Streptococcus sobrinus bacterial strain CBS 350.71 into OMZ 176
Actinomyces viscosus DSM 43329
Fusobacterium nucleatum multiform subspecies DSM 20482 solution
The red B of algae (Sigma) device colorimeter CR-200 (Minolta) preparation hydroxy-apatite flag
Preparation hydroxyapatite (HA) sheet compresses in the sheet mould by the hydroxyapatite with 250mg, with about 5,900kg (13, pressure compression 000lbs) 5 minutes.600 ℃ of following sintering hydroxy-apatite flags 4 hours, use the sterile deionized water aquation at last then.Sterilization hydroxy-apatite flag was 180 ℃ of sterilization HA sheets two hours.The preparation mutan
The preparation mutan, by being 6.5 at pH, 37 ℃ (keeping constant), Ventilation Rate is to cultivate Streptococcus sobrinus CBS 350.71 in the culture medium of 75rpm, and culture medium includes following component: NZ--Case 6.5g/l yeast extract 6g/l (NH
4)
2SO
420g/lK
2PO
43g/l glucose 50g/lPluronic PE6100 0.1%
After 35 hours, add sucrose, making its final concentration is that 60g/l is to induce glucosyltransferase.Total fermentation time is 75 hours.Centrifugal and filter the supernatant of (aseptic) fermentation liquid.Add sucrose then and make its final concentration be 5% (regulating pH to 7.0), spend the night at 37 ℃ of following agitating solutions simultaneously with acetic acid.Filtering solution is collected in insoluble mutan on the Propex simultaneously, and with containing 1% sodium benzoate, pH is the deionized water thorough washing of 5 (regulating with acetic acid).At last, with insoluble mutan lyophilizing with grind.Dextranase activity (KDU) is measured
One thousand Novo glucanase unit (1KDU) refers to per hour destroy the required enzyme amount of glucosan that forms the reducing sugar that is equivalent to 1g maltose in the Novo Nordisk method of measuring glucanase.Assay method is based on following standard conditions: 40 ℃ of pH 5.4 of 20 minutes substrate glucosan 500 (Pharmacia) response time temperature
Holding rope about the detailed description of Novo Nordisk analytical method (AF 120) can obtain.Mutanase activity (MU) is measured
A mutan enzyme unit (MU) refers to that per minute discharges the required enzyme amount of 1 μ mol reducing sugar under standard conditions.40 ℃ of pH 5.5 of 15 minutes 1.5% mutan response time of standard conditions substrate temperature
Holding rope about the detailed description of Novo Nordisk analytical method (AF 180/1-GB) can obtain from Novo Nordisk A/S.BAN
TM(bacterial amylase Novo) active (KNU) (thousand Novo-α-Dian Fenmei units) measures
The mensuration of standard activity is with respect to the analytical standard under the following condition: substrate: right-nitrobenzophenone-α-D-Fructus Hordei Germinatus seven glucosides (pNP-G7) temperature: 37 ℃ of pH:7.1
Holding rope about the detailed description of Novo Nordisk analytical method (Novo Nordisk publication AF 215) can obtain.Fungamyl
TMActive (FAU) measures
An AMS unit (1FAU) refers to per hour decompose 5.26g starch (Merck under the standard method of measuring AMS based on following standard conditions of Novo Nordisk; Amylumsolubile Erg.B.6, lot number 9947275) used enzyme amount: substrate: 37 ℃ of pH:4.7 of soluble starch reaction time: 7-20 minute temperature
Holding rope about the detailed description of Novo Nordisk analytical method can obtain.Producing maltase activity (MANU) measures
One is produced maltogenic amylase Novo unit (MANU) and is defined in the used enzyme amount of per minute hydrolysis 1 μ mol maltotriose under the standard conditions.Hold the copy that rope can obtain this analytical method.AMG
TMActive (AGU) measures
A Novo amyloglucosidase unit, (AGU) be defined in the used enzyme amount of per minute hydrolysis 1 μ mol maltose under the following standard conditions: substrate: maltose temperature: 25 ℃ of pH:4.3,30 minutes (acetate buffer) response time
Holding rope about the detailed description (AF 22) of this analytical method can obtain.Transglucosidase activity (AGU) is measured
Utilize aforesaid mensuration AMG
TMActive method is measured the transglucosidase activity, and represents with identical unit (AGU).CGT enzymatic activity (KNU) is measured
The assay method of CGT enzyme KNU enzymatic activity is the change a little to Phadebas amylase check (Pharmacia) method.With Phadebas tablet (Phadebas
TMThe amylase check Pharmacia) is used as substrate.This substrate is crosslinked insoluble blue starch polymer, mixes with bovine serum albumin and buffering material.After suspending in water, starch is through the hydrolysis of enzyme, and then the blue fragment of generation.Under 60 ℃, pH is 6.2, and incubation was measured after 15 minutes in the 0.15nM calcium.The blue solution that obtains is at the absorbance at 620nm place, corresponding to enzymatic activity.
Enzymatic activity is compared with the activity of standard enzyme, and this enzymatic activity representation unit is identical with the active representation unit of standard enzyme simultaneously.Standard enzyme is Termamyl
TM(Novo Nordisk A/D, Denmark) utilizes potato starch to measure its amylolytic activity as substrate.This method is mixed starch/enzymatic solution sample with iodine solution after the reaction mutually based on the decomposition of enzyme to the potato starch after modifying.At first, be shown as black-and-blue, but become more and more shallow in diastatic process Smalt, and be transformed into brownish red gradually, with reactant and the comparison of stained glass standard substance.One thousand Novo α-Dian Fenmei unit (KNU) is defined in standard conditions (promptly 37 ℃+/-0.05; 0.0003M Ca
2+PH is 5.6) the following used enzyme amount of dextrinize 5.26g starch dry matter (Merck Amylum solubility).Activity is represented with following Novo unit (NU)/ml.
The mensuration of CGT enzymatic activity is by under 60 ℃, and pH is an incubation dilution enzyme and carrying out in substrate 4-10 minute in 6.0 the 10mM sodium citrate.Promozyme
TMActive (PUN) measures
An amylopectin enzyme unit Novo (PUN) is defined as being hydrolyzed amylopectin; Discharge reduced sugar, the reducing power that reduced sugar has is equivalent to the required enzyme amount of per minute 1 μ mol glucose under following standard conditions: substrate: 0.2% amylopectin temperature: 40 ℃ of pH:5.0 (0.05M citrate buffer) reaction time: 30 minutes
The detailed description of relevant this analytical method (AF 90) is held rope and can be obtained.Assessment plaque-inhibiting effect
Be used to assess the method (JP 2250816) of the method for removal plaque effect based on the Kao description.According to this method, the hydroxy-apatite flag, as mentioned above through after the sterilization, by at three kinds of oral microorganism (Streptococcus sobrinus, actinomyces viscosus and Fusobacterium nucleatum) bacterial strain exists to place in the brain-heart infusion medium that contains 0.2% sucrose (Difco) down with different enzyme and spends the night covering one deck biomembrane on it.
In order to check the effect of plaque-inhibiting, will appear at plaque on the hydroxy-apatite flag with the PBS solution (saline of phosphate-buffered) that contains the red B of 0.1% algae and dye and be redness.Red intensity (is called a
*) on colorimeter CR-200, measure.Maximum a
*Value is 60.Show red color intensity not enough (being the less of plaque appearance) less than this value.a
*Value is zero to show do not have redness (promptly not having plaque).Plaque suppresses effect and represents with relative value, based on untreated biomembranous a
*Value is 100%.The plaque-inhibiting effect of the amylolytic enzyme that EXAMPLE Example 1 is different
Three kinds of oral microorganisms, Streptococcus sobrinus, actinomyces viscosus and Fusobacterium nucleatum, respectively in the presence of various enzymes, anaerobism overnight incubation in the time of 137 ℃.The hydroxy-apatite flag is immersed in the culture broth in the nurturing period after coating the aseptic saliva of one deck, has therefore just formed oral biological film on sheet.After the cultivation, use the normal saline washing hydroxy-apatite flag of phosphate-buffered simply, contain among the PBS of the red B of 0.1% algae at 1ml subsequently and cultivated 1 minute, dye redness with the plaque that will appear on the hydroxy-apatite flag.Remove the red B solution of algae, with PBS developing sheet a few minutes.Sheet placed to spend the night in air dry.On colorimeter CR-200, measure red color intensity (a
*), and be untreated sheet relatively.
The prevention plaque effect of a large amount of different enzymes is as shown in table 1.
Table 1: the inhibition of selected amylolytic enzyme/prevention plaque activity
* significance,statistical, P＜0.01** significance,statistical, P＜0.001
| Enzyme | Active | Plaque intensity (%) |
| Do not have (contrast) | ????100 | |
| Maltogenase TM(Production by Bacteria maltogenic alpha-amylase enzyme) | ????200MANU/ml | ????36.8** |
| Fungamy TM(fungal alpha-amylase) | ????40FAU/ml | ????45.3** |
| From the deutero-transglucosidase of aspergillus niger | ????32AGU/ml | ????54.2** |
| Promozym TM(antibacterial amylopectase) | ????10PUN/ml | ????45.0* |
As seeing in the above table, on listed activity level, all obtained prevention/plaque-inhibiting effect significantly on the statistics with all amylolytic enzymes.Embodiment 2 amylolytic enzymes, the prevention plaque effect that the starch modification enzyme combines with mutanase and glucanase
Three kinds of oral microorganisms, Streptococcus sobrinus, actinomyces viscosus and Fusobacterium nucleatum are respectively in 1MU/ml mutanase and 1KDU/ml glucanase and following table in the presence of the listed various enzymes, 37 ℃ of following anaerobism overnight incubation.The hydroxy-apatite flag is immersed in the culture broth in the nurturing period after coating the aseptic saliva of one deck, so that form oral biological film on the hydroxy-apatite flag.After the cultivation, sheet with handling as the method for describing among the embodiment 1, is measured red intensity.Table 2: amylolytic enzyme and starch modification enzyme are with the activity of the inhibition/prevention plaque of mutanase and glucanase
*: all culture broth all comprise 1MU/ml mutanase and 1kDU/ml glucanase
| Enzyme | Active | Plaque intensity (%) |
| Contrast * | ?100 | |
| BAN TM(bacterial) | 12KNU/ml | ?42.7 |
| The CGT enzyme | 0.32KNU/ml | ?40.4 |
Just as can be seen, with BAN
TMUse with mutanase and glucanase with the CGT enzyme, with independent use mutanase and glucanase by comparison, obtained the plaque-inhibiting effect that improves.The bonded additional effect of CGT enzyme and mutanase and glucanase is especially remarkable.For BAN
TMWith the CGT enzyme, they are combined with mutanase and glucanase have synergism, owing in this check, use BAN separately
TMPerhaps the CGT enzyme can not reduce the formation of plaque.The pheron dose curve of embodiment 3 inhibition/prevention plaque effects
Three kinds of oral microorganisms, Streptococcus sobrinus, actinomyces viscosus and Fusobacterium nucleatum are respectively at various enzyme (Fungamyl
TM, Promozyme
TMAnd Maltogenase
TM, all are all from NovoNordisk A/S) exist down, 37 ℃ of following anaerobism overnight incubation.The hydroxy-apatite flag is immersed in the culture broth in the nurturing period after coating the aseptic saliva of one deck, so that form oral biological film on sheet.After cultivating, sheet is simply with the normal saline washing of phosphate-buffered, cultivates in containing the 1ml PBS solution of the red B of 0.1% algae then and dyes redness with the plaque that will appear on the hydroxy-apatite flag in 1 minute.Remove the red B solution of algae, with PBS developing sheet a few minutes.Sheet is placed on dried overnight in the air.Utilize colorimeter CR-200 to measure red intensity (a
*), thereby determine to remain in biomembrane on the sheet, with the value comparison of the value that obtains with untreated sample.Result such as accompanying drawing 1,2 are shown in 3.
Claims (22)
1. oral cavity health compositions, at least a amylolytic enzyme and/or at least a starch modification enzyme that comprise plaque-inhibiting and/or remove the plaque effective dose.
2. according to the oral cavity health compositions of claim 1, starch modification enzyme wherein is the CGT enzyme.
3. according to the oral cavity health compositions of claim 2, CGT enzyme wherein is to be derived by following bacterial strain: the bacterial strain of Bacillus autolyticus, the bacterial strain of bacillus cereus, the bacterial strain of Bacillus circulans, Bacillus circulans is had a liking for the bacterial strain of alkali mutation, solidify the bacterial strain of bacillus cereus, the bacterial strain of strong bacillus cereus, the bacterial strain of Bacillus halophilus, the bacterial strain of bacillus macerans, the bacterial strain of bacillus megaterium, the bacterial strain of Bacillus ohbensis, the bacterial strain of bacstearothermophilus, the bacterial strain of bacillus subtilis, the bacterial strain of Klebsiella Pneumoniae, hot anaerobic bacillus(cillus anaerobicus) belongs to the bacterial strain of strain, the dwell bacterial strain of hot anaerobic bacillus(cillus anaerobicus) of the bacterial strain of the hot anaerobic bacillus(cillus anaerobicus) of producing and ethanol, fin, the bacterial strain of pyrolysis clostridium amylobacter, the bacterial strain of the bacterial strain of clostridium thermosaccharolyticum or the hot anaerobic bacillus(cillus anaerobicus) of delivery in hot weather sulfur.
4. according to the oral cavity health compositions of claim 1, wherein the starch modification enzyme is a transglucosidase.
5. according to the oral cavity health compositions of claim 4, transglucosidase is wherein derived by aspergillus niger.
6. according to the oral cavity health compositions of claim 1, amylolytic enzyme wherein is a α-Dian Fenmei.
7. according to the oral cavity health compositions of claim 6, α-Dian Fenmei wherein is a bacterial.
8. according to the oral cavity health compositions of claim 6 or 7, α-Dian Fenmei wherein is selected from following thing group: bacillus subtilis derives and next α-Dian Fenmei; Separate the α-Dian Fenmei that the starch bacillus is derived and come; Bacstearothermophilus derives and next α-Dian Fenmei; Aspergillus oryzae is derived and next α-Dian Fenmei; And non-pathogenic microorganism is derived and next α-Dian Fenmei.
9. according to the oral cavity health compositions of claim 6, wherein α-Dian Fenmei is a fungal alpha-amylase.
10. according to the oral cavity health compositions of claim 1, amylolytic enzyme wherein is a debranching enzyme, particularly amylopectase.
11. the oral cavity health compositions according to claim 2 to 10 in arbitrary further comprises glucanase and/or mutanase.
12. according to the oral cavity health compositions of claim 11, mutanase is wherein derived from Trichoderma harzianum, and/or glucanase is derived from the bacterial strain of paecilomyces.
13. an oral cavity health protection product includes according to claim 1 to 12 the oral cavity health compositions in arbitrary.
14. according to the oral health products of claim 13, its form has: toothpaste, tooth gel, dentifrice, collutory, dentifrice composition, chewing gum, lozenge, dental floss or toothpick.
15. one kind is used for the method that plaque-inhibiting formed or removed plaque, comprise tooth and/or chewing gum are contacted with compositions with oral cavity health during claim 1-12 is arbitrary, perhaps with claim 13 or 14 in oral health products contact a period of time, to obtain plaque-inhibiting or to remove the effect of plaque.
16., include regularly and brush teeth with the oral health products of toothpaste in the claim 14 or tooth gel form according to the method for claim 15.
17., include the oral health products flushing oral cavity of using the collutory form in the claim 14 regularly according to the method for claim 15.
18. according to the method for claim 15, oral health products wherein is a gum formats.
19. at least a amylolytic enzyme is used for the application that plaque-inhibiting formed and/or removed the compositions for health care in plaque oral cavity in preparation.
20. according to using wherein amylolytic enzyme such as claim 1 to 5,11 or 12 arbitrary middle definition in the claim 19.
21. use at least a starch modification enzyme to be used for the application that plaque-inhibiting formed and/or removed the compositions for health care in plaque oral cavity in preparation.
22. according to the application in the claim 21, wherein starch modification enzyme such as claim 1,6 to 10,11 or 12 arbitrary middle definition.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DK119197 | 1997-10-17 | ||
| DK1191/1997 | 1997-10-17 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1275906A true CN1275906A (en) | 2000-12-06 |
Family
ID=8102004
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN98810170A Pending CN1275906A (en) | 1997-10-17 | 1998-10-16 | Plaque-inhibiting oral compositions |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP1023037A1 (en) |
| JP (1) | JP2001520179A (en) |
| CN (1) | CN1275906A (en) |
| AU (1) | AU734737B2 (en) |
| CA (1) | CA2305804A1 (en) |
| WO (1) | WO1999020239A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104188861A (en) * | 2014-08-26 | 2014-12-10 | 华南理工大学 | Complex enzyme mouthwash for eliminating dental plaque and preparation method thereof |
| CN104207958A (en) * | 2014-08-26 | 2014-12-17 | 华南理工大学 | Denture care solution containing complex enzyme and capable of effectively cleaning dental plaque and preparation method thereof |
| CN104207983A (en) * | 2014-08-26 | 2014-12-17 | 华南理工大学 | Denture care solution containing enzyme and pericarpium citri reticulatae extract and preparation method thereof |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2846881B1 (en) * | 2002-11-13 | 2006-11-03 | TOOTHPASTE COMPOSITION CONTAINING AN AMYLOLYTIC ENZYME | |
| EP2258836B1 (en) * | 2004-09-10 | 2016-05-04 | Novozymes North America, Inc. | Methods for preventing, removing, reducing, or disrupting biofilm |
| DE102004046116A1 (en) * | 2004-09-23 | 2006-04-06 | Henkel Kgaa | Pullulanases from psychrophilic organisms |
| DE102006001148B4 (en) * | 2006-01-06 | 2008-03-27 | Henkel Kgaa | Oral and dental care and cleaners with enzyme (s) |
| DE102006004079A1 (en) * | 2006-01-28 | 2007-08-09 | Henkel Kgaa | Oral and dental care and cleaners with enzymes |
| CA2859164A1 (en) | 2011-12-21 | 2013-06-27 | Colgate-Palmolive Company | Oral care compositions |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3194738A (en) * | 1962-03-14 | 1965-07-13 | American Chicle Co | Chewing gums and dentifrices containing enzymes |
| FR7314M (en) * | 1967-08-17 | 1969-10-06 | ||
| GB1284728A (en) * | 1968-10-23 | 1972-08-09 | Aspro Nicholas Ltd | Dental caries control |
| LU59502A1 (en) * | 1969-09-24 | 1970-02-26 | ||
| BE756289A (en) * | 1969-09-25 | 1971-03-01 | Blendax Werke Schneider Co | TOOTHPASTE |
| FR2132980A5 (en) * | 1971-04-02 | 1972-11-24 | Investigations Sci Pharm | Dental and buccal hygiene compsns - contg enzymatic complexes |
| US4740368A (en) * | 1985-12-11 | 1988-04-26 | Plevy Donald J | Amylase containing breath cleansing confection |
| DE3903348A1 (en) * | 1989-02-04 | 1990-08-30 | Henkel Kgaa | MOUTH AND TOOTH CARE WITH POLYSACCHARIDE COLLECTING ENZYMES |
| GB2309705B (en) * | 1996-01-30 | 1999-09-08 | Kukident Gmbh | Denture cleansing |
| EP1011700B1 (en) * | 1997-06-17 | 2002-09-04 | Novozymes A/S | An oral care composition comprising a bacillus pullulanase and a dextranase |
-
1998
- 1998-10-16 AU AU96213/98A patent/AU734737B2/en not_active Ceased
- 1998-10-16 CA CA002305804A patent/CA2305804A1/en not_active Abandoned
- 1998-10-16 CN CN98810170A patent/CN1275906A/en active Pending
- 1998-10-16 WO PCT/DK1998/000452 patent/WO1999020239A1/en not_active Application Discontinuation
- 1998-10-16 JP JP2000516642A patent/JP2001520179A/en active Pending
- 1998-10-16 EP EP98949952A patent/EP1023037A1/en not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104188861A (en) * | 2014-08-26 | 2014-12-10 | 华南理工大学 | Complex enzyme mouthwash for eliminating dental plaque and preparation method thereof |
| CN104207958A (en) * | 2014-08-26 | 2014-12-17 | 华南理工大学 | Denture care solution containing complex enzyme and capable of effectively cleaning dental plaque and preparation method thereof |
| CN104207983A (en) * | 2014-08-26 | 2014-12-17 | 华南理工大学 | Denture care solution containing enzyme and pericarpium citri reticulatae extract and preparation method thereof |
| CN104207983B (en) * | 2014-08-26 | 2017-08-25 | 华南理工大学 | A kind of denture care liquid containing enzyme and dried orange peel extracts and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| AU9621398A (en) | 1999-05-10 |
| JP2001520179A (en) | 2001-10-30 |
| CA2305804A1 (en) | 1999-04-29 |
| AU734737B2 (en) | 2001-06-21 |
| WO1999020239A1 (en) | 1999-04-29 |
| EP1023037A1 (en) | 2000-08-02 |
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| ASS | Succession or assignment of patent right |
Owner name: NUOWOQIMEIZI CO.,LTD. Free format text: FORMER OWNER: NOVO NORDISK A/S Effective date: 20021024 |
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