WO2002047615A2 - Oral compositions and use thereof - Google Patents
Oral compositions and use thereof Download PDFInfo
- Publication number
- WO2002047615A2 WO2002047615A2 PCT/US2001/049032 US0149032W WO0247615A2 WO 2002047615 A2 WO2002047615 A2 WO 2002047615A2 US 0149032 W US0149032 W US 0149032W WO 0247615 A2 WO0247615 A2 WO 0247615A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- acid
- derivatives
- flavonoid
- oral
- oral composition
- Prior art date
Links
Classifications
-
- 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
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/0216—Solid or semisolid forms
-
- 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
- A61K8/31—Hydrocarbons
-
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
-
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/342—Alcohols having more than seven atoms in an unbroken chain
-
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/36—Carboxylic acids; Salts or anhydrides thereof
-
- 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
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
-
- 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
- A61K8/60—Sugars; Derivatives thereof
- A61K8/602—Glycosides, e.g. rutin
-
- 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
- A61K8/63—Steroids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/167—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Definitions
- the present invention relates to oral compositions and their use for inhibiting the activity of surface-bound glucosyltransferase; treating or inhibiting dental caries, gingivitis, candidiasis, and/or denture stomatitis; inhibiting the accumulation of microorganisms on an oral surface; and treating or inhibiting aphthous ulcerations on an oral surface.
- GTFs Glucosyltransferase enzymes
- Dental plaque is essentially a biof ⁇ lm. Glucans promote the adherence and accumulation of cariogenic streptococci on the tooth surface, and play an essential role in the development of pathogenic dental plaque related to caries activity (Hamada and Slade, 1980; Schilling and
- GTF B which synthesizes a polymer of mostly insoluble ⁇ l,3-linked glucan
- GTF C which synthesizes a mixture of insoluble 1,3 -linked glucan and soluble ⁇ l,6-linked glucan
- GTF D which synthesizes ⁇ l,6-linked soluble glucan
- GTF Ss may also be involved in the development of dental plaque (Nyvad and Kilian, 1987; Nacca-Smith et al, 2000). S. sanguinis colonizes tooth surface early in plaque formation, and its GTF catalyzes predominantly ⁇ l,6-linked soluble glucan (Ceska et al., 1972). Enzymatically active GTFs are present in the soluble fraction of whole human saliva and are also incorporated into salivary pellicle that is formed on tooth surfaces (R ⁇ lla et al., 1983; Scheie et al., 1987).
- GTFs incorporated into an experimental pellicle demonstrate distinct physical and kinetic properties when compared to the same enzymes in solution; GTF C and D express enhanced enzymatic activity (Schilling and Bowen, 1988; Nacca-Smith et al., 1996; Nenkitaraman et al., 1995). A large proportion of the glucans synthesized by these surface-adsorbed GTFs is retained on the pellicle and may provide binding sites for mutans streptococci, contributing to the in situ formation of dental plaque (Schilling and Bowen, 1988; Schilling and Bowen, 1992; Nacca-Smith and Bowen, 1998).
- Propolis a resinous substance collected by Apis mellifera bees from various plant sources and mixed with secreted beeswax, is a multifunctional material used by bees in the construction, maintenance and protection of their hives (Burdock, 1998; Ghisalberti, 1979). Propolis is a non-toxic natural product with multiple pharmacological effects and a complex chemical composition (Burdock, 1998; Ghisalberti, 1979).
- flavonoids have been considered the main biologically active compounds in propolis (Amoros et al., 1992; Bonhevi et al., 1994; Ghisalberti, 1979).
- Propolis exhibits a wide range of biological activities, including antimicrobial, anti-inflammatory, anesthetic, and cytostatic properties (Burdock, 1998; Ghisalberti, 1979).
- the present invention is directed to overcoming these and other deficiencies in the art.
- a first aspect of the present invention relates to an oral composition including: an organoleptically suitable carrier and an amount of a terpenoid and a flavonoid, dispersed in the carrier, which is effective to prevent or treat dental caries, dental plaque formation, gingivitis, candidiasis, dental stomatitis, aphthous ulceration, or fungal infections.
- a second aspect of the present invention relates to a method of inhibiting the activity of surface-bound glucosyltransferase which includes: contacting a surface-bound glucosyltransferase with an effective amount of a flavonoid or a combination of a flavonoid and a terpenoid, under conditions effective to inhibit the glucan-forming activity of the surface-bound glucosyltransferase.
- a third aspect of the present invention relates to a method of treating or inhibiting dental caries, gingivitis, candidiasis, or denture stomatitis, which method includes: providing an oral composition of the present invention and contacting an oral surface with an effective amount of the oral composition under conditions effective to treat or inhibit dental caries, gingivitis, candidiasis, or denture stomatitis.
- a fourth aspect of the present invention relates to a method of inhibiting accumulation of microorganisms on an oral surface which includes: providing an oral composition of the present invention and contacting an oral surface with an effective amount of the oral composition under conditions effective to inhibit accumulation of a microorganism which promotes dental caries, gingivitis, candidiasis, denture stomatitis, or formation of dental plaque matrix.
- a fifth aspect of the present invention relates to a method of treating or inhibiting aphthous ulceration which includes: contacting an oral surface with an effective amount of a terpenoid, a flavonoid, or a combination thereof, under conditions effective to treat an existing aphthous ulceration or inhibit formation of an aphthous ulceration.
- oral compositions of the present invention are particularly well suited for use in treating or inhibiting dental caries, gingivitis, candidiasis, and denture stomatitis, because oral compositions including terpenoids and flavonoids can both disrupt the activity of glucosyltransferases which are in solution and/or bound to a solid surface as well as destroy microorganisms which produce the glucosyltransferases. They are similarly well suited to inhibit accumulation of microorganisms which promote dental caries, gingivitis, candidiasis, denture stomatitis, or formation of dental plaque matrix. Additional benefits include minimizing the staining of teeth and oral malodors.
- Figure 1 is a graph illustrating time-kill curves for mutans streptococci strains by tt-farnesol at four times the MIC (or MBC). Streptococcus mutans GS-5 ( / ⁇ ), S. mutans UA 159 (•/o), S. sobrinus 6715 (win).
- FIGS 2A-B are graphs which illustrate the effects of apigenin on the activities of streptococcal GTFs in solution (2A) and adsorbed onto saliva-coated hydroxyapatite (sHA) surface (2B).
- GTF B ⁇
- GTF C ⁇
- GTF D ⁇
- GTF Ss o
- the data shown are mean values ( ⁇ SD).
- the percentage of inhibition was calculated considering the control (DMSO:EtOH, final concentration of 7.5% and 1.25%, vol./vol.) as maximum GTF activity.
- DMSO:EtOH final concentration of 7.5% and 1.25%, vol./vol.
- Figure 3 is a graph illustrating the effects of apigenin (1.33 mM) on the activities of GTFs in solution and adsorbed onto saliva-coated hydroxyapatite surface. Open bars represent solution assays and marked bars represent surface assays. The percentage of inhibition was calculated considering the control (solute) as 100% GTF activity. //-Farnesol (1.33 mM), chlorhexidine (1.33 mM), and fluoride (250 ppm) showed either negligible or moderate inhibitory effects.
- Figure 4 is a graph illustrating the effects of 1.33 mM tt-Farnesol and 0.12% CHX (1.33mM) on the viability oi Streptococcus mutans UA 159 biofilms. A similar profile was obtained for S. sobrinus 6715. Apigenin (1.33 mM) and fluoride (250 ppm) showed negligible antibacterial activity as measured by the killing of bacteria.
- Figure 5 is a graph illustrating the effects of treatments on smooth- surface caries and severity scores. Values capped by symbols are statistically significantly different from control (p ⁇ 0.05). ANONA, comparison for all pairs using Tukey-Kramer HSD.
- Figure 6 is a graph illustrating the effects of treatments on sulcal caries and severity scores. Values capped by symbols are statistically significantly different from control (p ⁇ 0.05). A ⁇ ONA, comparison for all pairs using Tukey-Kramer HSD.
- the present invention relates to oral compositions and their use in inhibiting the activity of surface-bound glucosyltransferase; treating or inhibiting dental caries, gingivitis, candidiasis, and/or denture stomatitis; inhibiting the accumulation of microorganisms on an oral surface; and treating or inhibiting aphthous ulcerations on an oral surface.
- Suitable terpenoids include, without limitation, terpenes, terpinols, diterpenic acids, diterpenes, triterpenes, and derivatives thereof.
- Exemplary terpenoids include, without limitation, tt-farnesol as well as its stereoisomers and derivatives, ⁇ -caryophyllene, terpineol, nerolidol, bisabolol, santatol, dehydroabietic acid, abietic acid, ⁇ -amyrine, triterpenic alcohol of amyrine, lanosterol, cupressic acid and its derivatives, agathic acid and its derivatives, agathalic acid, betuletol, melliferone, moronic acid, anwuweizonic acid, betulonic acid, syringaldehyde, imbricatoloic acid, communic acid, methyl isocupressate, tremetone, viscidone and its derivatives, ⁇ -ca
- the terpenoid is present in an amount which is effective to prevent or to treat dental caries, dental plaque formation, gingivitis, candidiasis, dental stomatitis, aphthous ulcerations, and/or fungal infections.
- the effective amount of terpenoid present in the oral composition is less than about 5 percent by weight/volume.
- the terpenoid is present in an amount which is between about 0.01 to about 2 percent by weight/volume, more preferably about 0.01 to about 1.5 percent by weight/volume.
- Suitable flavonoids include flavones, flavonols, dihydroflavonols, flavonones, and derivatives thereof.
- Exemplary flavonoids include, without limitation, apigenin and its derivatives, acacetin, baicalein, chrysin, luteolin, tectochrysin, kaempferol, kaempferide, galangin, isorhamnetin, rhamnetin, myricetin, fisetin, rutin, pinobanksin, pinobanksin-3 -acetate, pinobanksin-7-methyl eter, pinocembrin, sakuranetin, isosakuranetin, quercetin, hesperitin, naringin, pinostrobin and its derivatives, trihydroxymethoxy flavanone, tetraxydroxy flavanone, tetrahydroxyflavone, ermanin, 3,5,7-trihydroxy-4'-meth
- the flavonoid is present in an amount which is effective to inhibit soluble or surface-bound glucosyltransferase activity or to prevent or to treat dental caries, dental plaque formation, gingivitis, candidiasis, dental stomatitis, aphthous ulcerations, and/or fungal infections.
- the effective amount of flavonoid present in the oral composition is less than about 5 percent by weight/volume.
- the flavonoid is present in an amount which is between about 0.01 to about 1 percent by weight/volume, more preferably about 0.01 to about 0.5 percent by weight/volume.
- the oral compositions can take the form of a toothpaste or gel, a powder, a solution (e.g., mouthwash or dental rinse), a suspension, an emulsion, a lozenge, a mucoadhesive vehicle, a tablet or a gum.
- the composition can also be presented in a delivery vehicle such as a dental floss impregnated with a composition of the present invention.
- the particular choice of carrier will depend, at least in part, upon the desired form which the oral composition will take.
- the carrier is preferably both organoleptically suitable and pharmaceutically acceptable for oral administration.
- the carrier will include as a major component one or more of the following: water, glycerin, alcohols such as ethanol, sorbitol, propylene glycol, etc., DMSO, curable polymers, and powders such as starch.
- water When water is employed, deionized water is preferred. Typically, water can comprise from about 10 to about 85 weight percent of the oral composition, depending upon the formulation.
- Polymeric delivery vehicles can include copolymers of polyvinylmethylether with maleic anhydride and other similar delivery enhancing polymers.
- the oral composition includes effective amounts of both a terpenoid and a flavonoid dispersed in an organoleptically suitable non-polymeric carrier (i.e., substantially free from other propolis components).
- the terpenoid, flavonoid, and non-polymeric carrier can be of the type described above, present in the amount described above.
- the molar ratio of terpenoid.-flavonoid can be between about 0J to about 10:1, preferably about 0.5 to about 5:1.
- the oral composition can include a number of additives, including without limitation, an abrasive agent, a gelling agent, a humectant, a cariostatic agent, a flavoring agent or sweetener, a desensitizing agent, an anti- calculus agent, a whitening agent, a surfactant, a binding agent, a preservative, an opacifying agent, a coloring agent, a buffering agent, or combinations thereof.
- additives including without limitation, an abrasive agent, a gelling agent, a humectant, a cariostatic agent, a flavoring agent or sweetener, a desensitizing agent, an anti- calculus agent, a whitening agent, a surfactant, a binding agent, a preservative, an opacifying agent, a coloring agent, a buffering agent, or combinations thereof.
- Abrasive agents are typically employed in dentifrice compositions. Suitable abrasive agents include, without limitation, aluminum oxide, aluminum hydroxide, calcium hydrogen phosphate dihydrate or anhydride, silica gel, zirconosilicate, silicic anhydride, aluminosilicate, calcium carbonate, calcium pyrophosphate, aluminum silicate, insoluble sodium metaphosphate, magnesium tertiary phosphate, magnesium carbonate, calcium sulfate, synthetic resins, and combinations thereof. Abrasives can generally be employed in effective amounts of between about 20 to about 90 weight percent, more typically about 20 to about 60 weight percent for dentifrices.
- Gelling agents can be used in various compositions to assist in processing. Suitable gelling agents include, without limitation, carrageenan, sodium carboxymethyl cellulose, alkali metal alginates such as sodium alginate, gums, polyvinyl alcohol, and vee gum or the like. Typically, the gelling agents are employed in amount of about 0.3 to about 5 weight percent.
- Humectants can also be employed in the oral compositions, particularly toothpastes and gels and oral rinses. Suitable humectants include sorbitol, glycerin, propylene glycol, 1,3-butylene glycol, polyethylene glycol, xylitol, maltitol, lactitol, or the like. The humectant can also be used as the bulk carrier in many instances, in which case it can be present in an amount of about 5 to about 90 weight percent, more typically about 10 to about 60 weight percent.
- Cariostatic agents i.e., non-flavonoid cariostatic agents
- Suitable cariostatic agents include, without limitation, sodium fluoride, stannous fluoride, aminefluorides, sodium monofluorophosphate, sodium trimeta-phosphate, triclosan, casein, or combinations thereof. If desired, the cariostatic agent can be present in an amount between about 0.01 to about 3 weight percent, more typically between about 0.02 to about 1 weight percent. Flavoring agents are desired in most oral compositions to enhance the flavor and palatability of the oral composition and, thus, the likelihood of their use.
- Suitable flavoring agents can be flavoring oils (e.g., oil of spearmint, peppermint, wintergreen, sassafras, clove, sage, eucalyptus, cinnamon, lemon, and orange, methyl salicylate, etc.) or sweeteners (e.g., sucrose, sucralose, lactose, maltose, xylitol, sodium cyclamate, perillartine, aspartyl phenyl alanine methyl ester, saccharine, etc.). Flavoring agents can be present, either individually or collectively, in an amount of about 0J to about 10 weight percent, more typically about 0J to about 5 weight percent.
- flavoring oils e.g., oil of spearmint, peppermint, wintergreen, sassafras, clove, sage, eucalyptus, cinnamon, lemon, and orange, methyl salicylate, etc.
- sweeteners e.g., suc
- Desensitizing agents can be introduced in some embodiments of the oral composition to treat individuals whose teeth are sensitive to thermal shock, chemicals, etc. Suitable desensitizing agents include, without limitation, potassium citrate, potassium chloride, potassium tartrate, potassium bicarbonate, potassium oxalate, potassium nitrate, and strontium salts. Desensitizing agents can be present, either individually or collectively, in an amount of about 0.1 to about 5 weight percent, more typically about 0.1 to about 3 weight percent.
- Anti-calculus agents can be introduced in some embodiments of the oral composition to treat tartar formation. Suitable anti-calculus agents include, without limitation, alkali-metal pyrophosphates, hypophosphite-containing polymers, organic phosphonates, phosphocitrates, and combinations thereof. Anti-calculus agents can be present, either individually or collectively, in an amount of about 0J to about 5 weight percent, more typically about 0.1 to about 3 weight percent.
- Whitening agents can be employed in some forms of the oral composition. Suitable whitening agent including urea peroxide, calcium peroxide, and hydrogen peroxide. Whitening agents can be employed in amounts of about 0.5 to about 5 weight percent. Surfactants can also be employed in the various oral compositions.
- anionic surfactants include, without limitation, sodium lauryl sulfate, sodium lauroyl sarcosinate, ⁇ -olefin sulfonate, taurate, lauryl monoglyceride sulfate, lauryl monoglyceride sulfonate, and combinations thereof.
- nonionic surfactants include, without limitation, TWEEN, lauroyl diethanol amide, stearyl monoglyceride, sucrose fatty acid esters, lactose fatty acid esters, lactitol fatty acid esters, maltitol fatty acid esters, polyoxyethylene sorbitan monostearate, and combinations thereof.
- exemplary ampholytic surfactants include, without limitation, betain and amino acid type surfactants.
- Surfactants can be present in amount of about 0.5 to about 15 weight percent, more typically about 0.5 to about 10 weight percent.
- Binding agents can be utilized, typically for tablet or lozenge forms. Such binding agents include sodium carboxymethyl-cellulose, xanthan gum, gum arabic, etc. as well as synthetic polymers such as polyacrylates and carboxyvinyl polymers. Binders can be present in amounts of about 0.5 to about 50 weight percent depending on the form of the oral composition.
- Preservatives can be utilized to enhance the storage properties of the oral composition.
- One suitable preservative is benzoate (e.g., sodium benzoate), which also possesses a degree of cariostatic activity.
- Titanium dioxide is a white powder which adds opacity to the compositions. Titanium dioxide can be present in an amount of about 0.25 to about 5 weight percent.
- Coloring agents may also be added to the oral compositions of the present invention. The coloring agent may be in the form of an aqueous solution, i.e., an approximately 1 percent coloring agent in water solution. Color solutions can be present in an amount of about 0.01 to about 5 weight percent.
- the oral composition may also include buffers and salts to buffer the pH anionic strength of the oral composition, thereby promoting its stability.
- the pH of such oral compositions of the invention is generally in the range of about 4.5 to about 9 or 10, preferably about 6.5 to about 7.5 or 8.
- the pH can be controlled with acid (e.g. citric acid or benzoic acid) or base (e.g. sodium hydroxide) or buffered (as with sodium citrate, benzoate, carbonate, or bicarbonate, disodium hydrogen phosphate, sodium dihydrogen phosphate, etc.).
- Preparation of the oral compositions of the present invention can be carried out according to known techniques and procedures, depending upon the particular type of vehicle employed.
- Suitable surfactants can be employed to enhance the solubility of the active ingredients in the selected carrier. Discussion of the preparation of oral compositions is presented in Harry's Cosmeticology, Seventh Edition, 1982, edited by J. B. Wilkinson and R. J. Moore, published by Chemical Publishing of New York, pages 609 to 617, which is hereby incorporated by reference in its entirety.
- the oral composition are to be packaged according to conventional procedures.
- a toothpaste or dental cream or gel dentifrice as well as a dental gel will usually be in a collapsible tube or in a squeeze, pump or pressurized dispenser for metering out the contents, having a label describing it, in substance, as a toothpaste, dental cream or the like.
- a mouth rinse will generally be in a glass or plastic bottle.
- Lozenges and gum will be packaged individually or in blister packages as is known in the art.
- the composition is preferably applied regularly to oral surfaces, such as every day or every second or third day or preferably from two to three times daily, for at least two weeks up to eight weeks or more up to lifetime.
- the oral compositions find numerous uses for improving oral health or inhibiting the decline of oral health.
- these aspects of the present invention involve treating or inhibiting dental caries, gingivitis, candidiasis, denture stomatitis, and/or aphthous ulceration, as well as inhibiting accumulation of microorganisms on an oral surface.
- these aspects of the present invention are carried out by contacting an oral surface (e.g., a tooth, gum or other mucosal surface, tongue surface, a surface on partial or complete dentures, etc.) with an effective amount of an oral composition of the present invention under conditions effective to achieve the desired effect (i.e., treat or inhibit the above conditions).
- an oral surface e.g., a tooth, gum or other mucosal surface, tongue surface, a surface on partial or complete dentures, etc.
- a number of oral conditions can be treated, including dental caries, gingivitis, candidiasis, denture stomatitis, and aphthous ulcerations.
- Dental caries is one of the most prevalent and significant forms of oral disease which can lead to loss of teeth in spite of development of therapeutics.
- dental caries results in tooth loss as a result of organic acids, the natural metabolite of plaque bacteria contained in dental plaque, which decalcify dental hard tissue locally and gradually.
- fluoride treatments have provided some improvement in public health, there still exists a need to improve the prevention of dental caries.
- Bacterial plaque on tooth surfaces is also a major etiological factor in gingivitis and periodontitis. Recently, specific oral microorganisms in plaque have been implicated in chronic periodontal disease.
- One approach to mitigate developing gingivitis and periodontitis is to prevent or reduce plaque formation.
- the present invention offers an improved approach for the treatment and inhibition of gingivitis by treating oral surfaces with an oral composition of the present invention.
- Candidiasis and denture stomatitis are two conditions which are particularly concerned with Candida invasion. Oral candidiasis is seen frequently in two populations, patients wearing dentures and patients who are immunosuppressed or have AIDS.
- compositions of the present invention offer an attractive option to treat or inhibit candidiasis and denture stomatitis.
- Aphthous ulcerations are painful oral lesions occurring on the mucous membranes of the tongue, lips, cheek, soft and hard palates, gingiva, floor of the mouth, or pharynx.
- aphthous stomatitis which is manifested by the formation of aphthous ulcers.
- the disease is still not completely understood, and the factors which cause aphthous stomatitis are still being investigated. Regardless of their cause, the ulcers typically last from 5 to 21 days. Often the ulcers form in groups and merge into a singular extensively ulcerated area.
- the lesions may become extremely painful, especially during periods of fatigue or during eating, which may become difficult if not impossible. Because of the ability of oral compositions of the present invention to inhibit accumulation of microorganisms on an oral surface, the degree of infection at the site of an aphthous ulcer can be reduced, thereby treating the aphthous ulcer or inhibiting its formation in the first place.
- the oral compositions of the present invention are particularly well suited for use in treating or inhibiting dental caries, gingivitis, candidiasis, denture stomatitis, and/or aphthous ulceration, because terpenoids and flavonoids, particularly when used in combination, can disrupt the activity of glucosyltransferases which are in solution or bound in a solid surface as well as destroy microorganisms which produce the glucosyltransferases.
- the oral compositions of the present invention are also well suited to inhibit accumulation of microorganisms which promote dental caries, gingivitis, candidiasis, denture stomatitis, or formation of dental plaque matrix.
- Exemplary microorganisms, the accumulation of which can be inhibited include without limitation: lactobacilli, actinomyces, leptotrichiae, non- ⁇ -hemolytic streptococci, enterococci, miscellaneous gram-positive cocci, neisseriae, diphtheroid bacilli, fusiform bacilli, bacteroides, spirochetes, yeasts (Candida), and combinations thereof.
- Another use of the oral compositions of the present invention concerns the inhibition of surface-bound glucosyltransferases. It is believed that the present invention for the first time discloses the novel use of flavonoids to inhibit the activity of surface-bound glucosyltransferase, which exhibit different behavior as compared to soluble glucosyltransferase.
- This aspect of the present invention can be achieved by contacting a surface-bound glucosyltransferase with an effective amount of a flavonoid or a combination of a flavonoid and a terpenoid, under conditions effective to inhibit the glucan-forming activity of the surface-bound glucosyltransferase.
- Glucosyltransferases which can be inhibited include, without limitation, S. mutans GTF B, S. mutans GTF C, S. mutans GTF D, S. sobrinus GTF, and S. sanguinis GTF.
- Test compounds The compounds used in this study are classified into three groups: 1) flavonoids (flavonols, flavones, flavanones and dihydroflavonols); 2) cinnamic acid derivatives; and 3) terpenoids.
- flavonols kaempferide, kaempferol, galangin, isorhamnetin, rhamnetin, myricetin, fisetin, ratin
- flavones apigenin, acacetin, baicalein, chrysin, luteolin, tectochrysin
- flavanones pinocembrin, sakuranetin, isosakuranetin
- cirmamic acid derivatives ferulic acid, p-coumaric acid, caffeic acid
- terpenoids tt-farnesol, ⁇ -caryophyllene, terpineol, syringaldehyde
- Protocatechuic acid, vanillin, chlorhexidine (CHX), sodium fluoride, and benzoic acid were obtained from Sigma-Aldrich Co., Mass.
- Bacterial strains The bacterial strains used for the production of GTFs were: 1) Streptococcus mille ⁇ KSB8, which harbors the gtfB gene from S. mutans GS-5 (for GTF B production); 2) S. milleri NH5, which contain the gtfD gene S. mutans GS-5 (for GTF D); 3) S. mutans WHB 410 (Wunder and Bowen, 1999), which the genes for GTF B, D and fructosyltransferase were deleted (for GTF C); and 4) S. sanguinis 10904.
- the following bacterial strains were used: 1) S. mutans GS-5; 2) S.
- S. mutans UA159 were used for antibacterial assays: 1) S. mutans UAl59; and 2) S. sobrinus 6715.
- the cultures were stored at-80°C in brain heart infusion (BHI) or tryptic soy broth (TSB) containing 20% glycerol.
- BHI brain heart infusion
- TTB tryptic soy broth
- the S. milleri constructs were a kind gift from Dr. Howard K. Kuramitsu (SUNY, Buffalo, NY) and S. mutans UA 159 was obtained from Dr. Robert E. Marquis (Univ. of Rochester, Rochester, NY).
- GTFs B, D, and Ss were obtained from culture supernatants and purified to near homogeneity by hydroxyapatite column chromatography as described by Venkitaraman et al. (1995) and Vacca Smith et al. (2000). For GTF C isolation from S.
- mutans WHB 410 (Wunder and Bowen, 1999), cell pellets were harvested from low molecular weight broth (2.5% tryptone, 1.5% yeast extract, 0.3% glucose, 0.1% fructose and 0.1% sorbitol, which had been ultrafiltered through a 10-kd molecular weight cut-off membrane) cultures of S. mutans WHB 410 (ftf- gtfD- gtfB- derivative of S. mutans UA 130) grown in dialysis tubing (Schilling and Bowen, 1988). The cells were washed twice in 20 mM potassium phosphate buffer, pH 7.5 containing lrnM PMSF, and 0.02% sodium azide.
- the cells were then resuspended in 30 ml of 50 mM potassium phosphate buffer, pH 7.5 containing 0.1% triton X-100, 2M urea, 500 mM NaCl, 0.02% sodium azide and ImM PMSF, and incubated at 25°C for 2h under gentle agitation.
- the cell suspension was centrifuged at 10,000 g for 15 min. at 4 °C.
- the supernatant was carefully collected as the source of GTF C and dialyzed against 50 mM potassium buffer, pH 7.5 containing ImM PMSF and 0.02% sodium azide.
- the dialyzed preparation was purified by hydroxyapatite column chromatography as detailed by Venkitaraman et al. (1995).
- the purity of the enzyme preparations was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in a Hoefer Mighty Small SE245 system (Hoefer Scientific Instruments, San Francisco, CA, USA) and silver staining (Morrisey, 1981). Pre-stained standards were purchased from BioRad Laboratories. Protein concentration was determined by the method of Lowry et al. (1951), with bovine serum albumin (Sigma Chemical Co., St. Louis, MO) used to construct standard curves.
- Glucosyltransferase activity was measured by the incorporation of [ 14 C - glucose] from labeled sucrose (NEN Research Products, Boston, Mass.) into glucans (Germaine et al., 1974; Venkitaraman et al, 1995).
- the GTF enzyme added to each sample for all assays was equivalent to the amount required to incorporate 1.0- 1.5 ⁇ mol of glucose over the four hours reaction (1.0-1.5 units).
- Example 1 Effect of Terpenoids and Flavonoids on GTF Activity in Solution and Adsorbed onto Saliva-coated Hydroxyapatite Surface Assay
- the GTFs were adsorbed to hydroxyapatite beads coated with clarified whole saliva (free of GTF activity), as detailed elsewhere (Koo et al., 2000c; Schilling and Bowen, 1988, Venkitaraman et al., 1995).
- the saliva- coated hydroxyapatite (sHA) beads were exposed to sufficient enzyme to saturate the surface as determined experimentally. Following the adsorption of the enzyme, the beads were washed 3 times with buffer to remove the loosely bound material and exposed to 300 ⁇ l of the two-fold dilution series of test compounds (or control) for 30 min. at the same concentrations described above.
- the beads were washed and exposed to 300 ⁇ l [ I4 C-glucose]-sucrose substrate (100.0 mmol/1 sucrose, final concentration).
- the radiolabeled glucan formed was collected and quantified by scintillation counting (Germaine et al, 1974; Venkitaraman et al., 1995). All the solution and surface assays were done in quadruplicate in at least 3 different experiments. The effects of the most active compounds on the activity of GTFs are shown in the Tables 2 and 3 below. In general, flavonols and flavones reduced the activity of all the enzymes tested in solution (40 to 95% inhibition) and surface (15 to 60% inhibition) at concentration of 500 ⁇ M.
- apigenin (a 5,7,4' - trihydroxyflavone) displayed the most potent inhibition of GTFs activities.
- Apigenin inhibited 90.5 to 95% of the activity of all GTFs in solution at a concentration as low as 500 ⁇ M (135 ⁇ g/ml).
- the inhibitory effect of apigenin on surface adsorbed enzymes was not as potent as that observed when the same enzymes were in solution. Nevertheless, it was an effective inhibitor (30 to 60% inhibition at a concentration of 500 ⁇ M).
- the inhibitory effects of apigenin on GTFs are illustrated in Fig. 1.
- Apigenin reduced the activity of all enzymes in solution in direct proportion to the amount added in the reaction test (r 2 values ranging from 0.92 to 0.99).
- the IC 50 of apigenin (the concentration of test compound required to inhibit the enzymatic activity by 50%) for the GTFs in solution was between 58 (16 ⁇ g/ml) to 98 ⁇ M (26 ⁇ g/ml).
- the IC 50 for surface-adsorbed enzymes were noticeably higher; the IC 50 values for GTF B and C were 478 (128 ⁇ g/ml) and 458 ⁇ M (122 ⁇ g/ml); those for GTF D and SS were >lmM. It is conceivable that an IC 50 for this agent would not be achieved for surface-adsorbed GTFs D and Ss.
- the surface-adsorbed GTF B and C were inhibited significantly more at all concentrations tested than GTF D and Ss (p ⁇ 0.05).
- the flavone baicalein, and the flavonols myricetin and rhamnetin also showed to be effective inhibitors of GTFs in solution (70-90% at 500 ⁇ M) and adsorbed on saliva- coated hydroxyapatite surface (19-40% at 500 ⁇ M).
- inhibitory curves concentration-activity; concentrations ranging from 62.5 ⁇ M to 1 mM
- IC 50 values concentration of test compounds required to inhibit the enzymatic activity by 50%
- a one-way layout experimental design was used in 4x2x7 factorial scheme (enzyme x state x dose). An analysis of variance was carried out and qualitative treatments were compared using Tukey test at level of 5% of significance (p ⁇ 0.05). A non-linear regression was applied in order to evaluate effects of different concentrations. The results are summarized in Figures 2A-B.
- the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for each test compound according to the National Committee for Clinical Laboratory Standards guidelines (Tentative standard M26-T, 1992; NCCLS Publication No. M7-A5, 2000) and Koo et al. (2000b).
- the broth microdilution and macrodilution methods (in TSB) were used for the antibacterial tests.
- the starting inoculum was 5 x 10 5 CFU/ml and the concentrations of test compounds ranged from 15.6 to 500 ⁇ M (two-fold dilutions).
- the MICs and MBCs were determined in quadruplicate in at least 3 different experiments.
- the flavanones, dihydroflavonols, and some terpenoids (tt-farnesol and ⁇ -caryophyllene) tested in this study showed moderate inhibitory effects (8-45% for GTFs in solution and 7-24% for GTFs on surface at a concentration of 500 ⁇ M); the cinnamic acid derivatives showed negligible effects on GTF enzymes. In some cases the activity of GTFs was enhanced, e.g. by cinnamic acid derivatives and some terpenoids (e.g. protocatechuic acid, terpineol).
- the MIC and MBC values of the test compounds for S. mutans (GS-5 and UA 159) and S. sobrinus 6715 are shown in Table 4 below.
- Some of the flavanones and dihydroflavonols, as well as tt-farnesol (terpenoid) displayed antibacterial activity. All flavanones inhibited bacterial growth, among them pinocembrin was the most effective with MIC values of 250 ⁇ M (64 ⁇ g/ml) for all strains tested. Pinocembrin showed bactericidal effect against S. sobrinus 6715 at 500 ⁇ M (128 ⁇ g/ml).
- the dihydroflavonol pinobanksin-3 -acetate also inhibited the growth of S.
- sobrinus 6715 and S. mutans strains (MIC values of 500 ⁇ M or 157 ⁇ g/ml). Among all test compounds, tt-farnesol was the most effective antibacterial agent. The MIC values were 125 ⁇ M (28 ⁇ g/ml) for S. mutans strains and 62.5 ⁇ M (14 ⁇ g/ml) for S. sobrinus 6715. The MBC values were 500 ⁇ M (112 ⁇ g/ml) for S. mutans strains and 250 ⁇ M (56 ⁇ g/ml) for S. sobrinus 6715.
- Chlorhexidine (positive control) yielded MIC values between 1.1 -2.2 ⁇ M (1 -2 ⁇ g/ml) and MBC values of 8.9 ⁇ M (8 ⁇ g/ml). Flavonols, flavones and cinnamic acid derivatives did not show any antibacterial activity at concentrations used in this assay, with exception to baicalein (MIC values of 500 ⁇ M).
- Chlorhexidine (positive control) showed MIC values between 1.1-2.2 ⁇ M (1-2 ⁇ g/ml) and MBC value of 8.9 ⁇ M (8 ⁇ g/ml).
- the diluted sample (50 ⁇ l) was plated on to tryptic soy agar by means of a spiral plater (Autoplate model 3000, Spiral Biotech, Inc., Bethesda, Md.). The plates were incubated in 5% CO 2 at 37 °C for 48h, when the number of colonies was determined. Killing curves were constructed by plotting log ⁇ 0 CFU/ml versus time over 24h. All the assays were done in quadruplicate on at least 3 occasions.
- Bactericidal effect was defined as a > 3 log 10 decreases in the CFU/ml from the original inoculum.
- the potential for drug carry-over to produce falsely low viability counts was minimized by dilution of inocula and plating of small volumes of diluted samples (50 ⁇ l).
- no evidence of drug carry-over was detected at the lowest dilution used for plating (10 "1 ).
- propolis reduced dental caries in desalivated rats (Koo et al., 1999).
- the present invention was directed to the identification of specific compounds in propolis that can, alone and/or in combination, inhibit growth of cariogenic bacteria and the activity of the GTFs, which are associated with the pathogenesis of dental caries and other oral conditions. This was the first step toward identifying novel inhibitors of GTF enzymes and mutans streptococci growth.
- Apigenin a 4', 5, 7-trihydroxyflavone, was the most effective inhibitor of GTFs, especially GTF B and C.
- apigenin greatly inhibited GTF, especially GTF B and C, irrespective of whether the enzyme was exposed before or after adsorption to a surface at concentration as low as 500 ⁇ M. This level of inhibition has not been observed previously (Vacca-Smith and Bowen, 1997; Wunder and Bowen, 1999).
- apigenin The effective inhibition of GTF B and C by apigenin may affect the pathogenic potential of dental plaque related to caries, consistent with a reduction in smooth-surface caries observed with mutants of mutans streptococci defective in the production of either or both GTFs (Yamashita et al., 1993).
- Apigenin is a non-mutagenic flavonoid displaying a variety of anti- tumor and anti-inflammatory effects (Liang et al., 1999; McVean et al., 2000).
- the above data is believed to be the first demonstration of apigenin as a potent inhibitor of GTFs activity.
- the resistance displayed by surface-adsorbed GTF enzymes may be related to conformational changes the GTF undergoes during the adsorption process, consistent with differences in physical/kinetic properties and products synthesized between adsorbed and soluble forms of the enzyme (Kopec et al., 1997; Schilling and Bowen, 1988; Venkitaraman et al., 1995).
- biofilms of S. mutans UA159 and S. sobrinus 6715 were used for time-kill studies. Biofilms were formed on standard glass microscope slides in batch cultures for 5 days (Curran et al., 1998). Cells of mutans streptococci were grown in tryptone-yeast extract broth with addition of 1% (w/v) sucrose at 37°C and 5% CO 2 . Typically, 5-day-old biofilms yield approximately 10 9 colony forming units (CFU) per slide. The killing assays were performed according to Phan et al. (2000).
- the homogenized suspension was serially diluted (10 1 to 10 4 ) and plated on tryptic soy agar or blood agar by means of a spiral plater (Autoplate model 3000, Spiral Biotech, Inc., Bethesda, Md). The plates were incubated in 5% CO 2 at 37 °C for 48h, when the number of colonies was determined. Killing curves were constmcted by plotting log i0 CFU/ml versus time over 4h. All the assays were carried out in quadraplicate on at least 3 different occasions. Bactericidal effect was defined as a > 3 log 10 decreases in the CFU/ml from initial viable counts, at time zero.
- apigenin on the activity of glucosyltransferases (GTFs) are shown in Fig. 3.
- Apigenin at 1.33 mM (0.035%) is a potent inhibitor of GTFs whether the enzyme is in solution (90-95% inhibition) or on a surface (60-70%); the effects of tt-farnesol on enzyme activity were negligible (10-20%).
- Chlorhexidine (CHX) showed only moderate effects (30-45% inhibition in solution and 10-20% on surface).
- both tt-farnesol and CHX showed antibacterial activity against S. mutans and S. sobrinus biofilms as illustrated in Fig. 4.
- tt-Farnesol at 1.33 mM reduced the viable counts of mutans streptococci, showing 1 log 10 decrease in CFU/ml after 2-4h incubation.
- CHX at 1.33mM (0.12%) was more effective in reducing the viability of biofilms than was tt-farnesol (2-3 log 10 decrease in CFU/mL after same period of incubation). None of the antibacterial agents tested was completely bactericidal against mutans streptococci biofilms. Apigenin was devoid of antibacterial effects against mutans streptococci.
- the lower left jaw was aseptically dissected, suspended in 5.0 mL of sterile saline solution, and sonicated (three 10-second pulses with 5-second intervals at 30 watts, Branson Sonifier 450).
- the suspension was plated on mitis salivarius agar plus streptomycin to estimate S. sobrinus population and on blood agar to determine total cultivable flora (TCF).
- Dental caries was evaluated according to Larson's modification of Keyes' system (Larson, 1981). The determination of caries score was blind by codification of the jaws and was done by 1 calibrated examiner.
- the rats remained in apparent good health during the 5-week experiment. Weight gains were not significantly different among the treatment groups (p>0.05).
- the effects of the treatments on the incidence and severity of smooth surface caries are shown in Fig. 5.
- the tt-farnesol + apigenin (60% reduction, Keyes's score: 3.3+3 J), fluoride (72%, 2.3+3.0) and chlorhexidine (75%, 2.0+1.9) groups significantly reduced the incidence of smooth surface caries compared to control group (p ⁇ 0.05).
- the smooth-surface caries severity scores were significantly lower in the groups treated with apigenin, apigenin+tt-farnesol, fluoride, chlorhexidine (Ds and Dm levels) and tt-farnesol (Dm) than control (p ⁇ 0.05).
- the incidence and severity of sulcal-surface caries were reduced by fluoride and chlorhexidine treatments only (p ⁇ 0.05) as shown in Fig. 6.
- the combination of apigenin and tt-farnesol showed significantly better results than individual compounds when compared to control (p ⁇ 0.05).
- the percentage of S. sobrinus recovered from the jaws of the rats was calculated from the total cultivable flora and S. sobrinus population.
- Glucosyltransferases synthesize glucan, a capsular-like material, which can protect microorganisms from inimical influences such as antibacterial or antifungal agents, e.g., famesol. It is believed that the prevention of glucan formation enhances the accessibility of antibacterial or antifungal agents such as famesol to contact the microorganisms which they are intended to effect, thereby enhancing their efficacy.
- tt-farnesol was able to reduce the severity of smooth- surface caries at Ds level.
- Terpenes such as famesol have been reported to disrupt membrane function, ultimately reducing cell viability (Bard et al., 1988).
- a high concentration of tt-farnesol ⁇ 10 mM would be needed to disrupt the streptococci cells membrane (Koo et al., 2001).
- the combination of apigenin and tt-farnesol was more cariostatic than either of the compounds alone.
- CHX and fluoride effectively reduced the incidence of smooth-surface and sulcal surface caries, thereby supporting the validity of the selected model.
- the anti-plaque effect of CHX has been largely attributed to its antimicrobial activity and oral substantivity; it is a cationic substance that binds to soft and hard tissues of the mouth, as well as to bacterial cell walls (Rolla and Melsen, 1975; Jones, 1997).
- CHX reduced the viability of mutans streptococci biofilms (Fig. 4), although it was not bactericidal. This data confirms the findings in the animal experiment where lower levels of S. sobrinus infection were detected in the group treated with CHX.
- Fluoride 250 ppm was devoid of detectable antimicrobial and anti-GTF activity as measured here, although it may reduce acid tolerance and acid production of S. mutans (Marquis, 1990; Belli et al., 1995). Nevertheless, fluoride is the most effective anti-caries agent known to date (Rolla et al., 1991; Clarkson et al., 2000). There is a consensus that the main effect of fluoride is to interfere physicochemically with caries development by reducing demineralization and enhancing remineralization of dental enamel (Dawes and ten Gate, 1990).
- apigenin could be such an agent.
- the above data show clearly that two natural compounds from propolis were cariostatic in the animal model, even at low concentrations. Apigenin was shown to be the first natural cariostatic agent based on its ability to inhibit GTFs; it is a promising anti-caries compound which has a distinct mechanism of action compared to other clinically proven agents.
- Streptococcus mutans GS-5 Can. J. Microbiol. 41(9):785-791. Bonhevi et al., 1994. The composition, active components and bacteriostatic activity of propolis in dietetics. J. Am. Oil. Chem. Soc. 71:529-532. Bowen et al., 1988. The effect of desalivation on coronal and root surface caries in rats. J. Dent. Res. 67(l):21-23. Budtz-Jorgensen, 1974. The significance of Candida albicans in denture stomatitis.
- Streptococcus mutans J. Dent. Res. 53:1355-1360. Ghisalberti, 1979. Propolis: A review. Bee World 60:59-84. Gilbert et al., 1997. Biof ⁇ lm susceptibility to antimicrobials. Adv. Dent. Res. l l(l):160-7. Hamada and Slade, 1980. Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol. Rev. 44:331-384. Hanada and Kuramitsu, 1988. Isolation and characterization of the Streptococcus mutans gtfC gene, coding for synthesis of both soluble and insoluble glucans.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Emergency Medicine (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Cosmetics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP01991293A EP1399119A2 (en) | 2000-12-13 | 2001-12-13 | Oral compositions and use thereof |
AU2002231030A AU2002231030A1 (en) | 2000-12-13 | 2001-12-13 | Oral compositions and use thereof |
JP2002549191A JP2004521880A (en) | 2000-12-13 | 2001-12-13 | Oral composition and method of using the same |
CA002431044A CA2431044A1 (en) | 2000-12-13 | 2001-12-13 | Oral compositions and use thereof |
US10/450,231 US20040057908A1 (en) | 2001-12-13 | 2001-12-13 | Oral compositions and use thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US25530400P | 2000-12-13 | 2000-12-13 | |
US60/255,304 | 2000-12-13 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2002047615A2 true WO2002047615A2 (en) | 2002-06-20 |
WO2002047615A3 WO2002047615A3 (en) | 2003-12-31 |
Family
ID=22967716
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2001/049032 WO2002047615A2 (en) | 2000-12-13 | 2001-12-13 | Oral compositions and use thereof |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP1399119A2 (en) |
JP (1) | JP2004521880A (en) |
AU (1) | AU2002231030A1 (en) |
CA (1) | CA2431044A1 (en) |
WO (1) | WO2002047615A2 (en) |
Cited By (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003051125A1 (en) * | 2001-12-18 | 2003-06-26 | Henkel Kommanditgesellschaft Auf Aktien | Adhesion inhibition of fungi |
WO2006068973A2 (en) | 2004-12-22 | 2006-06-29 | Colgate-Palmolive Company | Methods for use of oral care compositions containing free-b-ring flavonoid anti-oxidants |
WO2006069210A3 (en) * | 2004-12-22 | 2006-11-09 | Colgate Palmolive Co | Oral care compositions containing free-b-ring flavonoids and flavans |
WO2006045056A3 (en) * | 2004-10-19 | 2007-02-01 | Unigen Pharamaceuticals Inc | Flavonoid composition for treating oral diseases |
EP1845964A1 (en) * | 2004-12-20 | 2007-10-24 | Kimberly-Clark Worldwide, Inc. | Antimicrobial compositions comprising a natural agent selected from gallic acid, eucalyptol, naringin, a jasmonic acid compound and any combination thereof |
WO2008009956A1 (en) | 2006-07-20 | 2008-01-24 | Oraldent Limited | Combinations for oral compositions, their preparation and use |
EP1973515A1 (en) * | 2005-12-21 | 2008-10-01 | Colgate-Palmolive Company | Oral compositions comprising propolis |
WO2008144942A1 (en) * | 2007-05-31 | 2008-12-04 | F.P.L. Pharma Inc. | Sesquiterpene formulations, kits and methods of use thereof |
US7695743B2 (en) | 2002-04-30 | 2010-04-13 | Unigen Pharmaceuticals, Inc. | Formulation of a mixture of Free-B-Ring flavonoids and flavans for use in the prevention and treatment of cognitive decline and age-related memory impairments |
EP2279662A3 (en) * | 2006-03-10 | 2011-04-13 | Yuuzou Tsuchida | 5,7,4'-trihydroxy-3',5'-dimethoxyflavone as an antibacterial agent |
EP2368442A2 (en) | 2005-07-27 | 2011-09-28 | Symrise AG | Use of hesperetin for enhancing the sweet taste |
TWI404545B (en) * | 2004-12-22 | 2013-08-11 | Colgate Palmolive Co | Oral care compositions containing flavonoids and flavans |
US8568799B2 (en) | 2002-03-22 | 2013-10-29 | Unigen, Inc. | Isolation of a dual COX-2 and 5-lipoxygenase inhibitor from acacia |
US8771761B2 (en) | 2006-10-12 | 2014-07-08 | Unigen, Inc. | Composition for treating atopic dermatitis comprising extracts of bamboo and scutellaria |
WO2014163512A1 (en) * | 2013-04-05 | 2014-10-09 | Manuka Health New Zealand Limited | Therapeutic compositions comprising extracts of propolis and uses thereof |
US9061039B2 (en) | 2002-03-01 | 2015-06-23 | Unigen, Inc. | Identification of Free-B-Ring flavonoids as potent COX-2 inhibitors |
CN104721066A (en) * | 2013-12-19 | 2015-06-24 | 高露洁-棕榄公司 | Rutin solubilising method using polyhydroxy alkyl alcohol |
WO2015143575A1 (en) * | 2014-03-28 | 2015-10-01 | Universidad De Concepcion | Tissue-conditioning kit comprising a terpenic anti-fungal agent exhibiting activity against oral candidiasis present in subprosthetic stomatitis |
WO2016010440A1 (en) * | 2014-07-18 | 2016-01-21 | Manuka Health New Zealand Limited | Propolis and extracts thereof for the treatment of skin cancers and improvement of skin health |
WO2016044297A3 (en) * | 2014-09-15 | 2016-05-12 | Vizuri Health Sciences Llc | Polyphenol/flavonoid compositions and methods of formulating oral hygienic products |
US9370544B2 (en) | 2002-04-30 | 2016-06-21 | Unigen, Inc. | Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent |
WO2017048617A1 (en) * | 2015-09-15 | 2017-03-23 | Vizuri Health Sciences Llc | Polyphenol/flavonoid compositions and methods of formulating oral hygienic products |
US9622964B2 (en) | 2003-04-04 | 2017-04-18 | Unigen, Inc. | Formulation of dual cycloxygenase (COX) and lipoxygenase (LOX) inhibitors for mammal skin care |
RU2688231C1 (en) * | 2019-02-19 | 2019-05-21 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермский государственный медицинский университет имени академика Е.А. Вагнера" Министерства здравоохранения Российской Федерации | Method of treating leptopothia in the oral cavity |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8945518B2 (en) | 2002-04-30 | 2015-02-03 | Unigen, Inc. | Formulation of dual eicosanoid system and cytokine system inhibitors for use in the prevention and treatment of oral diseases and conditions |
HUE057104T2 (en) | 2004-01-23 | 2022-04-28 | Eden Research Plc | Methods of killing nematodes comprising the application of a terpene component |
AP2919A (en) | 2004-05-20 | 2014-05-31 | Eden Research Plc | Compositions containing a hollow glucan particle or a cell wall particle encapsulating a terpene component, methods of making and using them |
GB0514943D0 (en) * | 2005-07-20 | 2005-08-24 | Botan Ltd | Oral care compositions |
JP2007091706A (en) * | 2005-08-30 | 2007-04-12 | Kao Corp | Biofilm inhibitor |
FI121468B (en) * | 2006-06-07 | 2010-11-30 | Valtion Teknillinen | Compounds derived from betulin as antimicrobial agents |
KR101409266B1 (en) * | 2011-11-24 | 2014-06-24 | 영남대학교 산학협력단 | Inhibitors of Biofilm and Methods therefor |
KR101685671B1 (en) * | 2012-09-27 | 2016-12-12 | 한국 한의학 연구원 | Pharmaceutical composition for preventing or treating peridontal disease comprising extract of Lignum Acronychiae containing nerolidol |
KR101597645B1 (en) * | 2014-02-12 | 2016-02-25 | 재단법인 진안홍삼연구소 | A pharmaceutical composition containing acacetin as a active ingredient for the treatment of dental caries and peridontal disease |
Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4382886A (en) * | 1981-04-13 | 1983-05-10 | Sosnowski Zenon M | Method for extracting propolis and water soluble dry propolis powder |
US4748022A (en) * | 1985-03-25 | 1988-05-31 | Busciglio John A | Topical composition |
US5116602A (en) * | 1989-09-27 | 1992-05-26 | Colgate-Palmolive Company | Antiplaque oral compositions |
US5472684A (en) * | 1993-06-02 | 1995-12-05 | Colgate Palmolive Company | Oral compositions for plaque and gingivitis |
US5529779A (en) * | 1991-08-27 | 1996-06-25 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Purified propolis-extract, and its preparation and uses |
US5561116A (en) * | 1991-04-11 | 1996-10-01 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Solid product containing propolis components, and preparation and uses thereof |
US5922324A (en) * | 1995-01-31 | 1999-07-13 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Propolis extract with improved water-solubility |
US6027716A (en) * | 1997-04-02 | 2000-02-22 | Farmo-Nat Ltd. | Synergistic herbal extracts |
US6153227A (en) * | 1998-07-31 | 2000-11-28 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Propolis extract |
US6159450A (en) * | 1996-10-25 | 2000-12-12 | Societe De Conseils De Recherches Et D'applications Scientifiques S.C.R.A.S. | Use of Ginkgo biloba flavonoidic extract substantially devoid of terpenes for oral hygiene and composition containing such extract |
-
2001
- 2001-12-13 EP EP01991293A patent/EP1399119A2/en not_active Withdrawn
- 2001-12-13 AU AU2002231030A patent/AU2002231030A1/en not_active Abandoned
- 2001-12-13 CA CA002431044A patent/CA2431044A1/en not_active Abandoned
- 2001-12-13 JP JP2002549191A patent/JP2004521880A/en active Pending
- 2001-12-13 WO PCT/US2001/049032 patent/WO2002047615A2/en not_active Application Discontinuation
Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4382886A (en) * | 1981-04-13 | 1983-05-10 | Sosnowski Zenon M | Method for extracting propolis and water soluble dry propolis powder |
US4748022A (en) * | 1985-03-25 | 1988-05-31 | Busciglio John A | Topical composition |
US5116602A (en) * | 1989-09-27 | 1992-05-26 | Colgate-Palmolive Company | Antiplaque oral compositions |
US5561116A (en) * | 1991-04-11 | 1996-10-01 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Solid product containing propolis components, and preparation and uses thereof |
US5529779A (en) * | 1991-08-27 | 1996-06-25 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Purified propolis-extract, and its preparation and uses |
US5472684A (en) * | 1993-06-02 | 1995-12-05 | Colgate Palmolive Company | Oral compositions for plaque and gingivitis |
US5922324A (en) * | 1995-01-31 | 1999-07-13 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Propolis extract with improved water-solubility |
US6159450A (en) * | 1996-10-25 | 2000-12-12 | Societe De Conseils De Recherches Et D'applications Scientifiques S.C.R.A.S. | Use of Ginkgo biloba flavonoidic extract substantially devoid of terpenes for oral hygiene and composition containing such extract |
US6027716A (en) * | 1997-04-02 | 2000-02-22 | Farmo-Nat Ltd. | Synergistic herbal extracts |
US6153227A (en) * | 1998-07-31 | 2000-11-28 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Propolis extract |
US6153228A (en) * | 1998-07-31 | 2000-11-28 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Propolis extract |
Cited By (44)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003051125A1 (en) * | 2001-12-18 | 2003-06-26 | Henkel Kommanditgesellschaft Auf Aktien | Adhesion inhibition of fungi |
US9061039B2 (en) | 2002-03-01 | 2015-06-23 | Unigen, Inc. | Identification of Free-B-Ring flavonoids as potent COX-2 inhibitors |
US9168242B2 (en) | 2002-03-22 | 2015-10-27 | Unigen, Inc. | Isolation of a dual COX-2 and 5-lipdxygenase inhibitor from Acacia |
US8568799B2 (en) | 2002-03-22 | 2013-10-29 | Unigen, Inc. | Isolation of a dual COX-2 and 5-lipoxygenase inhibitor from acacia |
US9849152B2 (en) | 2002-04-30 | 2017-12-26 | Unigen, Inc. | Formulation of a mixture of Free-B-ring flavonoids and flavans as a therapeutic agent |
US8034387B2 (en) | 2002-04-30 | 2011-10-11 | Unigen, Inc. | Formulation of a mixture of free-B-ring flavonoids and flavans for use in the prevention and treatment of cognitive decline and age-related memory impairments |
US9655940B2 (en) | 2002-04-30 | 2017-05-23 | Unigen, Inc. | Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent |
US7695743B2 (en) | 2002-04-30 | 2010-04-13 | Unigen Pharmaceuticals, Inc. | Formulation of a mixture of Free-B-Ring flavonoids and flavans for use in the prevention and treatment of cognitive decline and age-related memory impairments |
US8652535B2 (en) | 2002-04-30 | 2014-02-18 | Unigen, Inc. | Formulation of a mixture of free-B-ring flavonoids and flavans for use in the prevention and treatment of cognitive decline and age-related memory impairments |
US9370544B2 (en) | 2002-04-30 | 2016-06-21 | Unigen, Inc. | Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent |
US9622964B2 (en) | 2003-04-04 | 2017-04-18 | Unigen, Inc. | Formulation of dual cycloxygenase (COX) and lipoxygenase (LOX) inhibitors for mammal skin care |
WO2006045056A3 (en) * | 2004-10-19 | 2007-02-01 | Unigen Pharamaceuticals Inc | Flavonoid composition for treating oral diseases |
CN101083981B (en) * | 2004-10-19 | 2013-03-27 | 尤尼根公司 | Flavonoid composition for treating oral diseases |
EP1845964A1 (en) * | 2004-12-20 | 2007-10-24 | Kimberly-Clark Worldwide, Inc. | Antimicrobial compositions comprising a natural agent selected from gallic acid, eucalyptol, naringin, a jasmonic acid compound and any combination thereof |
TWI404545B (en) * | 2004-12-22 | 2013-08-11 | Colgate Palmolive Co | Oral care compositions containing flavonoids and flavans |
AU2005319184B2 (en) * | 2004-12-22 | 2011-10-06 | Colgate-Palmolive Company | Oral care compositions containing free-B-ring flavonoids and flavans |
AU2005319184C1 (en) * | 2004-12-22 | 2012-05-10 | Colgate-Palmolive Company | Oral care compositions containing free-B-ring flavonoids and flavans |
WO2006068973A3 (en) * | 2004-12-22 | 2007-01-04 | Colgate Palmolive Co | Methods for use of oral care compositions containing free-b-ring flavonoid anti-oxidants |
WO2006069210A3 (en) * | 2004-12-22 | 2006-11-09 | Colgate Palmolive Co | Oral care compositions containing free-b-ring flavonoids and flavans |
EP2308565A3 (en) * | 2004-12-22 | 2016-02-10 | Colgate-Palmolive Company | Oral care compositions containing free-b-ring flavonoids and flavans |
WO2006068973A2 (en) | 2004-12-22 | 2006-06-29 | Colgate-Palmolive Company | Methods for use of oral care compositions containing free-b-ring flavonoid anti-oxidants |
EP2368442A2 (en) | 2005-07-27 | 2011-09-28 | Symrise AG | Use of hesperetin for enhancing the sweet taste |
EP1973515A1 (en) * | 2005-12-21 | 2008-10-01 | Colgate-Palmolive Company | Oral compositions comprising propolis |
US9005680B2 (en) | 2005-12-21 | 2015-04-14 | Colgate-Palmolive Company | Oral compositions comprising propolis |
EP2279662A3 (en) * | 2006-03-10 | 2011-04-13 | Yuuzou Tsuchida | 5,7,4'-trihydroxy-3',5'-dimethoxyflavone as an antibacterial agent |
EP2043593B1 (en) * | 2006-07-20 | 2012-04-04 | OralDent Limited | Oral compositions, their preparation and use |
US9987214B2 (en) | 2006-07-20 | 2018-06-05 | Oraldent Limited | Combinations for oral compositions, their preparation and use |
WO2008009956A1 (en) | 2006-07-20 | 2008-01-24 | Oraldent Limited | Combinations for oral compositions, their preparation and use |
EP2043593A1 (en) * | 2006-07-20 | 2009-04-08 | OralDent Limited | Oral compositions, their preparation and use |
US9532939B2 (en) | 2006-07-20 | 2017-01-03 | Ricerfarma S.R.L. | Combinations for oral compositions, their preparation and use |
US9623068B2 (en) | 2006-10-12 | 2017-04-18 | Unigen, Inc. | Composition for treating atopic dermatitis comprising extracts of bamboo and scutellaria |
US8771761B2 (en) | 2006-10-12 | 2014-07-08 | Unigen, Inc. | Composition for treating atopic dermatitis comprising extracts of bamboo and scutellaria |
WO2008144942A1 (en) * | 2007-05-31 | 2008-12-04 | F.P.L. Pharma Inc. | Sesquiterpene formulations, kits and methods of use thereof |
WO2014163512A1 (en) * | 2013-04-05 | 2014-10-09 | Manuka Health New Zealand Limited | Therapeutic compositions comprising extracts of propolis and uses thereof |
CN105163728A (en) * | 2013-04-05 | 2015-12-16 | 蜜纽康新西兰有限公司 | Therapeutic compositions comprising extracts of propolis and uses thereof |
CN104721066A (en) * | 2013-12-19 | 2015-06-24 | 高露洁-棕榄公司 | Rutin solubilising method using polyhydroxy alkyl alcohol |
AU2014367213B2 (en) * | 2013-12-19 | 2017-04-13 | Colgate-Palmolive Company | Method for solubilizing rutin with polyhydroxy alkyl alcohols |
US9883999B2 (en) | 2013-12-19 | 2018-02-06 | Colgate-Palmolive Company | Method for solubilizing rutin with polyhydroxy alkyl alcohols |
WO2015094479A1 (en) * | 2013-12-19 | 2015-06-25 | Colgate-Palmolive Company | Method for solubilizing rutin with polyhydroxy alkyl alcohols |
WO2015143575A1 (en) * | 2014-03-28 | 2015-10-01 | Universidad De Concepcion | Tissue-conditioning kit comprising a terpenic anti-fungal agent exhibiting activity against oral candidiasis present in subprosthetic stomatitis |
WO2016010440A1 (en) * | 2014-07-18 | 2016-01-21 | Manuka Health New Zealand Limited | Propolis and extracts thereof for the treatment of skin cancers and improvement of skin health |
WO2016044297A3 (en) * | 2014-09-15 | 2016-05-12 | Vizuri Health Sciences Llc | Polyphenol/flavonoid compositions and methods of formulating oral hygienic products |
WO2017048617A1 (en) * | 2015-09-15 | 2017-03-23 | Vizuri Health Sciences Llc | Polyphenol/flavonoid compositions and methods of formulating oral hygienic products |
RU2688231C1 (en) * | 2019-02-19 | 2019-05-21 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермский государственный медицинский университет имени академика Е.А. Вагнера" Министерства здравоохранения Российской Федерации | Method of treating leptopothia in the oral cavity |
Also Published As
Publication number | Publication date |
---|---|
WO2002047615A3 (en) | 2003-12-31 |
EP1399119A2 (en) | 2004-03-24 |
JP2004521880A (en) | 2004-07-22 |
CA2431044A1 (en) | 2002-06-20 |
AU2002231030A1 (en) | 2002-06-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20040057908A1 (en) | Oral compositions and use thereof | |
WO2002047615A2 (en) | Oral compositions and use thereof | |
JP6650785B2 (en) | Oral care method and composition using chitosan derivative compound | |
JP2019137702A (en) | Oral formulation of polyglucosamine derivatives in combination with non-fermentable sugar | |
JP2006199661A (en) | Coaggregation inhibitor | |
TWI733123B (en) | Prebiotic oral care compositions containing an alkyl glycoside | |
TW201102060A (en) | Anti-biofilm carbonate compounds for use in oral care compositions | |
JP2007131601A (en) | Composition for oral cavity | |
JP5925431B2 (en) | Caries prevention agent | |
JP5631175B2 (en) | Oral composition | |
JP2001206830A (en) | Composition for oral cavity | |
JP4323980B2 (en) | Oral composition | |
KR20170103476A (en) | Composition for prevention or treatment of oral disease comprising ROSAE LAEVIGATAE Extract | |
JP3336688B2 (en) | Oral composition | |
JPS6388123A (en) | Novel agent for dental caries | |
JP5892716B2 (en) | Oral composition | |
US20240197589A1 (en) | Oral care compositions comprising geranylgeraniol and methods of use thereof | |
JP3821037B2 (en) | Periodontal pathogen adhesion inhibitor and composition for oral cavity having periodontal pathogen adhesion inhibitory action | |
AU2019100473A4 (en) | Methods and compositions for protecting and promoting healthy oral microbiome | |
KR20020082308A (en) | Composition for the mouth containing extraction of Euonymus alatus Sieb | |
WO2024204726A1 (en) | Oral composition containing lactobacillus sporogenes | |
KR0138247B1 (en) | The oral cavity hygienic compositions | |
AU2021351695A9 (en) | Prebiotic oral care compositions and methods | |
KR20170103482A (en) | Composition for prevention or treatment of oral disease comprising Glechoma hederacea extract | |
KR101818211B1 (en) | Composition for prevention or treatment of oral disease comprising Arctill fructus Extract |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TN TR TT TZ UA UG US UZ VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2431044 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2002549191 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2002231030 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2001991293 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 10450231 Country of ref document: US |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
WWP | Wipo information: published in national office |
Ref document number: 2001991293 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 2001991293 Country of ref document: EP |