MXPA98008768A - Stable dentirical composition in storage containing casein and glicomacropept - Google Patents

Abstract

The present invention relates to stable oral compositions containing an antibacterial casein glycomacropeptide, an anionic surfactant, a hydrolyzed protein stabilizer which stabilizes the casein glycomacropeptide in the presence of an anionic surfactant against inactivation without reducing the antibacterial properties

Description

STABLE DENTÍFRICA COMPOSITION IN STORAGE CONTAINING CASEIN AND GLYCOMACROPEPTIDE BACKGROUND OF THE INVENTION 1. Field of the Invention This invention is directed to a dentifrice composition and more particularly to a dentifrice composition containing a casein glycomacropeptide having antiplaque and anti-caries activities. 2. The prior art 15 Casein glycomacropeptides have been identified by the art as being effective antibacterial agents against the microorganisms responsible for dental plaque and caries. For example, the patents of United States of America Nos. 4,992,420 and 4,994,441 disclose that the casein phosphopeptides derived from appanatein are effective in inhibiting the growth of streptococcus mutans, a bacterial species associated with dental caries and plaque formation.

As an antibacterial agent, it is desirable to formulate oral compositions such as toothpastes, gels or mouth rinses, by means of which the glycoacryptide of casein is conveniently delivered to the oral tissue. However, efforts to utilize such casein glyco acidoptides in dental products suitable for home use, casein glycomacropeptides have been found to be incompatible with anionic surfactants, such as sodium lauryl sulfate conventionally used to prepare the 10 dentifrice compositions such as toothpastes and gels, the casein glycomacropeptide being biologically inactivated by the surfactant.

Therefore, there is a clear need in the art to prepare oral care products using casein glycomacropeptides, wherein ingredients such as the surfactants used to prepare the oral care product do not interact or otherwise inactivate the glycomacropeptide. of casein present in the composition, so that an optimal antibacterial efficacy of the glycomacropeptide is available when the composition is applied to the oral tissue.

SYNTHESIS OF THE INVENTION In accordance with the present invention, there is provided an oral care composition wherein the casein glycomacropeptide compounds incorporated therein encounter minimal biological inactivation and provide excellent anticaries and antiplaque performance, such composition comprising an aqueous carrier having incorporated therein is a casein glycomacropeptide, an anionic surfactant and an amount of a hydrolyzed protein compound effective to reduce or prevent biological inactivation of the casein glycomacropeptide.

The oral compositions containing the casein glycomacropeptide and the hydrolyzed protein compound are effective with respect to plaque and caries control without compromising the rheological and foam characteristics of the composition. 15 Description of Preferred Additions The casein glycomacropeptide compounds useful in the practice of the present invention include those identified in the patent of the United States of America No. 4,992,420 with kappa-casein glycomacropeptides and desialylated derivatives thereof.

The term "kappa-casein glycopeptide" includes within its meaning a casein glycopeptide, which is the water-soluble component emanating from the hydrolysis dß-kappa-casein with rennet and a caseino-glycopeptide obtained by the proteolysis of caseino-acroglicopeptide. The desialylated derivatives are obtained from casein glycopeptides by more or less complete elimination of the sialic acids, for example the N-acetyl neuraminic and N-glycolyl neuraminic acids, from the oligosacid chains of the casein glycopeptide.

The preparation of the kappa-caseino-glycopeptides is more fully described in the patent of the States United States of America No. 4,992,420 whose description is incorporated herein by reference. Other glycomacropeptide casein compounds useful in the practice of the present invention are the kappa-casein glycomacropeptides described in U.S. Patent No. 5,075,424 whose The description is also incorporated herein by reference.

The glycomacropeptide casein is incorporated into the oral composition of the present invention at a concentration of about 0.5 to about 10 percent by weight, and preferably at about 3.0 to about 8 percent by weight.

Hydrolyzed protein compounds useful for preserving the antibacterial activity of casein glycomacropeptides, in accordance with the practice of the present invention includes the hydrolyzed collagen proteins, specifically the positively charged hydrolysates containing high concentrations of basic amino acids obtained by the extraction of a partially hydrolyzed collagen faction and the isolation by exchange treatment of a anion with an anion exchange resin or a partially charged hydrolyzed collagen protein. Such hydrolysed collagen proteins are known in the art and are more fully described in U.S. Patent No. 4,391,798, the disclosure of which is incorporated herein by reference. Other useful hydrolyzed proteins include gelatin, obtained by boiling animal parts such as the skin, tendons and ligaments and such other proteinaceous compounds, such as polylysine, polyarginine and protamine sulfate, salts of polyvinyl pyridinium ammonium and poly N- (2- hydroxypropyl methacrylamide).

Hydrolyzed collagen protein compounds are preferred for use as stabilizers of the antibacterial activity of glycomacropeptide casein. Commercially available hydrolyzed collagen proteins include Crotein Q®, a quaternary derivative of a hydrolyzed collagen protein commercially available from Croda, Inc., of New York, New York. Crotein Q has a minimum pl of 9.5-10.5, is a free-floating, faded powder and its adapted name is hydrolyzed animal protein of esteartrimonio. The free amino groups in the protein molecule react with the # quaternary ammonium reagent to form the quaternized derivative which has multiple positive changes. As shown diagrammatically below, at a pH below 5.5, Crotein Q will exhibit a double positive charge, due to the protonation of the NH groups in the protein chain. At a pH of 9.5, the quaternary group of Crotein Q remains positively charged. 3) 3 15 Another example of a commercially available hydrolyzed collagen protein is the gelatin (food grade) available from the American Gelatin Company, a regular hydrolyzed collagen protein prepared by boiling animal parts, such as skin, tendons, ligaments and bones with Water. Above a pH of 5.5, the gelatin product is not charged. At a low pH of 5.5, gelatin (type B) exhibits a positive charge due to the protonation of the NH groups in the protein chain, as shown diaquatically below.

The use of from about 0.1 to about 2.5 percent by weight of the hydrolyzed protein compound in the preparation of the composition of the present invention is sufficient to inhibit any inactivating effect of the antibacterial efficacy of the glycomacropeptide casein caused by the presence of a anionic surfactant in the composition.

To prepare a dentifrice composition such as a toothpaste or gel, abrasives including conventional toothpaste abrasives such as finely divided silica, hydrated alumina, calcium carbonate, calcium bicarbonate and calcium phosphate, anhydrous and or hydrate) are incorporated in the composition at a concentration of from about 10 to about 30 weight percent based on the composition and preferably at a concentration of about 5 to about 25 weight percent. At these abrasive levels, the vehicle is composed of from about 25 to about 40 percent by weight of water, as well as a humectant such as glycerol, sorbitol, propylene glycol or mixtures * of them at a concentration of from about 25 to about 10 of 45 percent by weight and preferably from about 25 to about 35 percent by weight of the composition.

Thickeners or gelling agents are used in the preparation of toothpastes of the present The invention also includes silica thickeners, carboxymethyl cellulose, sodium carboxymethyl cellulose, locust bean gum, iota carrageenan, tragacanth gum, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, and alginates and are incorporated into the oral compositions of the invention. concentrations from about 0.2 to about 3.0 percent by weight.

The surfactants which may be included in the composition of the present invention include surfactants anions, such as the water-soluble salts of the higher alkyl sulfates or the sulfoacetate, such as sodium lauryl sulfate, sulfoacetate, lauryl sodium or other suitable alkyl sulfate or sulfoacetate having from 8 to 18 carbon atoms in the alkyl group; water-soluble salts of sulfated monoglycerides of higher fatty acids, such as sodium coconut sulfonate or other suitable sulfonated monoglyceride of a fatty acid of 10 to 18 carbon atoms; sodium lauryl phosphate salts of higher fatty acid amides, for example 12 to 16 carbon atom acids, with the lower aliphatic amino acids, for example taurine or sarcosine, or other amino acids of two to six carbon atoms, such as sodium-N-methyl-N-palmitoyl taurido, sarcosinates N-lauroyl-N-myristoyl and N-palmitoyl sodium; water-soluble salts of the esters of such fatty acids with isethionic acid or with glycerol monosulfate, such as the sodium salt of the monoglyceride monosulphates of hydrogenated coconut oil fatty acids; water-soluble salts of olefin sulfonates, for example alkyne sulfates or hydroxyalkane sulfonates or mixtures thereof having from 12 to 16 carbon atoms in the carbon chain of the molecule; and acid-soluble soaps of water higher fatty acids such as those of 12 to 18 carbon atoms, for example, coconut fatty acids.

Surfactants are included in the composition of the present invention at a concentration of about 0.5 to about 3.0 weight percent and preferably about 1.0 to about 2.5 weight percent.

Various other materials may be incorporated into the tooth preparations of the present invention such as flavoring agents, sweetening agents and coloring materials, such as dyes and pigments, which are incorporated into the tooth compositions of the present invention in amounts , which do not adversely affect the properties and characteristics desired in the dentifrice components. * 10 Any suitable sweetening or flavoring materials can be employed to formulate a flavor for the oral composition of the present invention. Examples of suitable flavoring constituents include flavor oils, e.g., peppermint, peppermint oils, piperita, ferrule, sassafras, clove, sage, as well as methyl salicylate. Suitable sweetening agents include lactose, maltose, sodium cyclamate and saccharin. Suitably, flavor and sweetener agents together comprise from about 0.01 to 5 percent or more of the composition. Preferably, the The amount of the flavoring agent is from about 0.5 to about 2.0 percent by weight and the sweetening agent is from 0.1 to 4 or 0.1 to 0.5 percent by weight (the latter rank being for artificial sweeteners, such as saccharin) .

Pigments such as TiOz and coloring materials which are generally commercially available food dye solutions, which are inert with respect to other ingredients of the new oral composition are included at a concentration of about 0.05 to about 2.0 by cent by weight. In the preparation of toothpastes such as toothpastes and gels, a pre-mix in water was formed in which the water-soluble ingredients are first dissolved followed by the insoluble ingredients in water if there are any.

If desired, the lipophilic components can be pre-mixed together and such pre-mix can be mixed with the pre-mixed hydrophilic, after which the water-insoluble particulate materials can then be mixed. The pH of the dentifrice is maintained at a neutral pH and preferably at a pH between 6.7 and 7.2. For example, the thickener is dispersed with water and humectants. The surfactant, the hydrolyzed protein compound, the casein glycomacropeptide, the abrasive, the sweetener, the flavor and the dye are then separately added and uniformly dispersed. The toothpaste is then properly aerated (for example empty) and packed. The addition and mixing of the ingredients was carried out in a low humidity environment and preferably under a vacuum of 20-30 inches and preferably 28-30 inches of mercury.

In addition to dentifrices such as toothpaste or gel compositions for an oral application containing the hydrolyzed protein compound and the casein glycomacropeptide. The oral composition of the present invention may be in any other convenient form such as that of a mouthwash, a spray, a tablet, a lozenge, a powder for the teeth, and a chewing gum.

A typical mouthwash or mouthwash prepared in accordance with the practice of the present invention contains the following ingredient in percent by weight based on the weight of the mouthwash or spray composition. fifteen twenty A typical dental powder prepared according to the practice of the present invention contains the following ingredients in percent by weight based on the weight of the composition of dental powder.

A typical pill formula contains the following ingredients, percent by weight, based on the weight of the total formula.

Suitable humectants include sorbitol and glycerin. Emulsifiers include nonionic polyoxyesters such as polyoxyethylene sorbitan fatty esters, for example, sorbitan monolaurate 20, polyoxyethylene commercially available as polysorbate 20 and Tween 20. Tablet lubricants include magnesium stearate.

The manufacture of the mouthwashes, sprays and dentifrices of the present invention is comparatively simple because, in general, there is little or no critical order of addition of the various ingredients present in such compositions. In the preparation of a mouthwash or spray, the ingredients are dissolved in water and / or alcohol.

A typical chewing gum may contain the following ingredients in the gum formula. 10 fifteen twenty The chewing gums and the tablets can be manufactured by procedures normally employed in the manufacture of such products, with the macroglicopeptide of Casein is usually added preferably near the end of the manufacturing process if heat is used (as to minimize subjection to elevated temperatures).

The following examples serve to provide a further appreciation of the invention but are not intended to restrict the effective scope of the invention in any way. All percentages through the description and clauses are percent by weight of the final composition, unless otherwise indicated and where the percentages will be a total of 100 percent ingredients in the final composition.

• Example 1 10 To determine the stabilizing effect of a hydrolysed protein stabilizer such as Crotein Q or gelatin on a kappa-casein glycomacropeptide in the presence of an anionic surfactant (sodium lauryl sulfate) in a In the composition of toothpaste, a composition containing these ingredients was prepared by adding thickeners to a premix of water and humectant at a slightly elevated temperature (e.g., from 35 to 60 ° C) with providing the ingredients to a creamy consistency or of gel. The additional ingredients were then added. The resulting toothpaste was then deaerated, the flavor was introduced and the toothpaste was packed into tubes. The toothpastes were adjusted to a pH of 6.7 with NaOH. The ingredients of the tubed toothpaste compositions are listed in the Table I given below and identified as toothpastes 1 and 2. For comparison purposes, the procedure of Example 1 was repeated except that a hydrolysed protein stabilizer was not included in the toothpaste composition. The composition of the comparative toothpaste identified as toothpaste 3 which also had a pH of 6.7 is also listed in Table I given below.

Table I * 10 fifteen twenty As the pH of the toothpastes listed in Table I is 6.7, the gelatin stabilizer is not charged and the Crotein Q stabilizer has a positive charge.

The stabilizing effect of the hydrolyzed protein on the kappa-casein or glycomacropeptide (CGMP) in the presence of • the anionic surfactant, of sodium lauryl sulfate, was determined by an aging test in which the tubed toothpastes were exposed to the heated air at 40 ° C for a period of 12 weeks (which is equivalent to storage at room temperature for a period of 2 years) and then the amount of kappa-casein or glycomacropeptide remaining in the toothpaste was determined, expressed as "by recovery percent. " The method used to determine the percent recovery of kappa-casein glycomacropeptide was by means of capillary electrophoresis that made a separation and identification by movement in the electric potential (mobility) dependent on the size and charge of the molecule tested. The percent recovery of the glycomacropeptide kappa-casein for aged toothpastes is record in Table II given below.

Table II CGMP Stability @ 40 ° C • The results recorded in Table II show the superior aging stability of the toothpastes 1 and 2 containing as the hydrolyzed protein stabilizer in gelatin or Crotein Q in an anionic surfactant / CGMP dentifrice according to this invention in comparison to a identical toothpaste which did not contain such a hydrolyzed protein stabilizer. In the absence of the hydrolyzed protein stabilizer, comparative toothpaste 3 aged at 12 weeks exhibited a low level of kappa-casein recovery glycomacropeptide (34 percent) that is believed to be due to the naturalization of kappa-casein glycomacropeptide by the anionic surfactant, sodium lauryl sulfate. In the presence, however of any gelatin or Crotein Q or gelatin, percent recovery of kappa-casein glycomacropeptide of toothpastes 1 and 2 aged at 12 weeks was unexpectedly higher (72-79 percent) since the stabilizer apparently interacted with sodium lauryl sulfate surfactant to prevent inactivation of the glycomacropeptide kappa-casein. To determine the effect on the biological activity of the glycomacropeptide kappa-casein in the presence of a hydrolysed protein stabilizer, the paste 1 containing 5.0% by weight of kappa-casein glycomacropeptide was diluted with Water at a ratio by weight of 1:20 and was used to treat the saliva-coated hydroxyapatite (SCHA) beads according to a procedure wherein the radiolabeled γ-labeled iodine kappa-casein glycomacropeptide (125,) was prepared with a commercially available Bolton-Hunter reagent (catalog ICN No. 65001), following the procedure established in the ICN catalog. The iodine-labeled glycomacropeptide kappa-casein was separated from the unreacted labeled reagent by size exclusion chromatography on Sephadex G10 in a 0.1 M phosphate buffer containing 0.1 percent gelatin and 0.1 percent acid. Hydroxyapatite beads coated with saliva were prepared by exposing the hydroxyapatite of the HPLC class (Bio-Rad Catalog No. 130-0420) to human saliva at 37 ° C overnight, and then washing and resuspending in buffer about 67 mg / ml . A duplicate series of glycomacropeptide kappa-casein solutions standard, were prepared as serial dilutions of pure glycomacropeptide kappa-casein in a phosphate buffer saturated with magnesium and calcium ions, varying from about 3400 μg / ml down to 0; similar tubes were prepared with different dilutions of two extracts that have been prepared with kappa-casein glycomacropeptide. All tubes then received 0.5 ml of a hydroxyapatite suspension coated with saliva and 0.05 ml of radiolabeled glycophopropeptide kappa-casein at about 30,000 dpm.

The tubes were incubated at 37 ° C for one hour, then the solid phase was separated by centrifugation and both phases * were counted. Toothpaste 1 was extracted containing glycomacropeptide kappa-casein by displacing the labeled kappa-casein glycomacropeptide-labeled hydroxyapatite with saliva in the same manner as pure glycomacropeptide-kappa-casein containing about 2.5 mg / ml of the peptide; comparative paste 3 failed to displace the labeled kappa-casein glycomacropeptide significantly.

The biological activity of kappa-casein glycomacropeptide in toothpaste 1 was recorded in the Table III given below, the biological activity being the percent of kappa-casein glycomacropeptide, measured as being bound to the hydroxyapatite coated with saliva, since the casein glycomacropeptides containing kappa-casein have a high affinity (percent binding activity) to the hydroxyapatite coated with saliva due to the hydrophilic residues, for example the sialic acid bound to glycoproteins, in the casein protein. This in vitro test for biological astivity has previously been determined to be correlates with the antibacterial activity in vivo.

For the contrast purposes of toothpaste 3 and toothpaste C, which was the same as toothpaste 3 but did not contain either, kappa-casein glycomacropeptide or a hydrolyzed protein stabilizer were also tested for binding activity. The binding activity of kappa-casein glycomacropeptide in toothpaste 3 and toothpaste C was also recorded in Table 3 below.

As a control, a series of pure glycomacropeptide kappa-casein solutions designated to A-F solutions were prepared containing varying concentrations of glycosymethyl kappa-casein. The binding activity of the glycomacropeptide kappa-casein present in control solutions A-F was also recorded in Table III given below.

Table III Biological activity of CGMP Dental Pulp Stabilized with Gelatine The results recorded in Table III show that the concentration of kappa-casein glycomacropeptide in the AF control solutions was increased, the percent binding activity of kappa-casein glycomacropeptide also increased showing that the binding activity was provided at the concentration of kappa-casein glycomacropeptide present in the solution. The toothpaste extract of the kappa-casein toothpaste glycomacropeptide # Gelatin (toothpaste 1) provided essentially the same agglutinating activity than the pure glycomacropeptide kappa-casein control solutions (E-F control) of a similar concentration indicating that the kappa-casein gomomacropeptide toothpaste 1 stabilized with gelatin had a biological activity equivalent to kappa-casein pure glycomacropeptide and is not inactivated by other ingredients of the toothpaste. Toothpaste extract from toothpaste 3 containing comparative glycomacropeptide kappa-casein that did not contain gelatin stabilizer exhibited an essentially reduced binding activity (20 percent) and the extract of toothpaste toothpaste C that did not contain the kappa-casein glycomacropeptide (toothpaste C) did not exhibit any kappa-casein agglutination activity glycomacropeptide. meaningful

Claims (26)

1. An oral composition comprising in a dental vehicle a casein glycomacropeptide in an amount effective for antibacterial activity, an anionic surfactant and a stabilizer of hydrolyzed protein compound in an amount effective to stabilize the antibacterial activity of said antibacterial activity of casein glycomacropeptide against inactivation due to the presence of anionic surfactant.

2. An oral composition as claimed in clause 1, characterized in that the hydrolyzed protein stabilizer is a hydrolyzed collagen protein.

3. The composition as claimed in clause 2, characterized in that the hydrolyzed collagen protein is a quaternary derivative of a hydrolyzed collagen protein.

4. The composition as claimed in clause 2, characterized in that the hydrolyzed collagen protein is food class gelatin prepared from parts of boiled animals.

# 5. An oral composition as claimed in clause 1, characterized in that the hydrolyzed protein stabilizer constitutes about 0.1 to 2.5 percent by weight of the composition.

6. An oral composition as claimed in clause 1, characterized in that the casein glycomacropeptide constitutes about 0.5 to 10 weight percent of the composition.

7. An oral composition as claimed in clause 1, characterized in that the casein glycomacropeptide is kappa-casein glycopeptide and the desialiated derivatives thereof.

8. An oral composition as claimed in clause 1, characterized in that the anionic surfactant is sodium lauryl sulfate.

9. An oral composition as claimed in clause 1, characterized in that it is in the form of a toothpaste containing about 30-75 weight percent of a water insoluble polishing agent.

10. An oral composition as claimed in clause 1, characterized in that it is in the form of an edor of 70-80 percent hydrated alumina.

11. An oral composition as claimed in clause 1, characterized in that it is in the form of an aqueous mouth rinse containing about 5-10 percent ethanol.

12. An oral composition as claimed in clause 1, characterized in that it is in the form of a chewing gum comprising about 20 and 30 percent of a gum base containing a natural synthetic elastomer filler.

13. An oral composition as claimed in clause 9, characterized in that the polishing agent is silica.

14. A method for preparing a storage stable oral oral composition containing a casein glycomacropeptide having anti-caries and anti-plaque efficacy which stabilizes against biological inactivation in the presence of an anionic surfactant, its method comprises adding to an aqueous vehicle a glycomacropeptide of 25 casein in an amount effective for antibacterial activity, an anionic surfactant and a stabilizer composed of a fecal protein to stabilize the antibacterial activity of the casein glycosypropeptide against inactivation, due to the presence of the anionic surfactant and then mixing of the glycomacropeptide, the hydrolyzed protein and the anionic surfactant to obtain the stable composition.

15. The method as claimed in clause 14, characterized in that the protein stabilizer Hydrolyzed is a hydrolyzed collagen protein.

16. The method as claimed in clause 15, characterized in that the hydrolyzed collagen protein is a quaternary derivative of collagen protein 15 hydrolyzed.

17. The method as claimed in clause 15, characterized in that the hydrolyzed collagen protein is a food grade protein prepared from 20 boiled animal parts.

18. The method as claimed in clause 14, characterized in that the hydrolyzed protein stabilizer constituted about 0.1 to 2.5 percent by weight 25 of the composition.

19. The method as claimed in clause 14, characterized in that the casein glycomacropeptide constitutes about 0.5 to 10 percent by weight of the composition.

20. The method as claimed in clause 1, characterized in that the casein glycomacropeptide is a kappa-casein glycopeptide and a sialylated derivative thereof.

21. The method as claimed in clause 14, characterized in that the anionic surfactant lauryl sodium sulfate.

22. The method as claimed in clause 14, characterized in that the composition is in the form of a toothpaste containing about 30-75 weight percent of a water insoluble polishing agent.

23. The method as claimed in clause 14, characterized in that the composition is in the form of a toothpaste containing about 70-80 percent hydrated alumina.

24. The method as claimed in clause 14, characterized in that the composition is in a form of an aqueous mouthpiece containing about 5-10 percent ethanol.

25. The method as claimed in 5 clause 14, characterized in that the composition is in the form of a chewing gum, which comprises about 20-30 percent of a gum base containing a synthetic or natural elastomer filler.

26. The method as claimed in clause 22, sarasterized because the polishing agent is similar.