Compositions and methods for promoting mineralization

Abstract

To provide a composition and use thereof for reducing the visibility of a hypomineralized dental surface or subsurface of in tooth enamel.SOLUTION: In one aspect, the present invention provides a method for reducing visibility of a hypomineralized dental surface or subsurface. The method comprises: (i) bringing the hypomineralized dental surface or subsurface into contact with a liquid composition having a pH of 6 or less and comprising at least 40% w / v phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); and (ii) subsequently raising the pH of the liquid composition applied to the hypomineralized dental surface or subsurface to about 9 or higher, thereby forming a gel in and / or on the hypomineralized dental surface or subsurface and thereby reducing the visibility of the hypomineralized dental surface or subsurface.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to Australian Provisional Application No. 2019900834 and Australian Provisional Application No. 2019903859, the contents of each of which are incorporated herein by reference in their entirety.

[0002] The present invention relates to compositions for use in conjunction with mineralization of tooth surfaces, particularly tooth enamel. Methods are also provided for mineralizing hypomineralized lesions (such as subsurface lesions) in tooth enamel caused by various means, including dental caries, dental erosion, and fluorosis. [Background technology]

[0003] Common causes of hypomineralized lesions are dental caries and fluorosis.

[0004] Caries usually occurs due to demineralization of dental hard tissues through the fermentation of dietary carbohydrates by dental pathogens in plaque. Furthermore, restored tooth surfaces may be susceptible to further caries around the restoration. Tooth erosion or decay is the loss of tooth mineral due to dietary or refluxed acids. Tooth sensitivity is caused by loss of the protective mineralized layer, cementum, exposing the dentinal tubules. Calculus is the unwanted deposit of calcium phosphate mineral on the tooth surface. Thus, all these conditions, caries, tooth erosion, tooth sensitivity, and calculus, are related to an imbalance in calcium phosphate levels.

[0005] Enamel fluorosis (mottling) has been recognized for nearly a century, but the etiologic role of fluoride was not identified until 1942. The characteristic appearance of fluorosis distinguishes it from other enamel disorders. The clinical features of fluorous lesions of the enamel (FLE) are a continuum ranging from thin opaque lines followed by circumferential striae to chalky white enamel. The presence of a relatively highly mineralized outer surface of enamel and a hypomineralized subsurface in fluorotic lesions stimulates the development of an incipient enamel "white spot" carious lesion. With increasing severity, both the depth of enamel involved in the lesion and the degree of hypomineralization increase. The development of fluorosis The disease is highly dependent on the dose, time, and timing of fluoride exposure and appears to be associated with increased serum fluoride concentrations. Chalky "white spot" lesions can also occur on children's developing teeth, such as after treatment with antibiotics or fever. These lesions represent areas of hypomineralized (i.e., too little mineralization) tooth enamel.

[0006] Depending on the severity of the lesion, fluorosis is clinically managed by repair, replacement, or microabrasion of the outer enamel. These procedures are insufficient because they involve the restoration or removal of tooth tissue. Treatments that can mineralize the hypomineralized enamel to achieve a natural appearance and structure are desirable.

[0007] A specific complex of casein phosphopeptides and amorphous calcium phosphate ("CPP-ACP", commercially available as Recaldent®) has been shown to remineralize enamel subsurface lesions in vitro and in situ.

[0008] WO 98 / 40406, in the name of The University of Melbourne (the entire contents of which are incorporated herein by reference), describes casein phosphopeptide-amorphous calcium phosphate complexes (CPP-ACP) and CPP-stabilized amorphous calcium fluoride phosphate complexes (CPP-ACFP) formed at alkaline pH. Such complexes have been shown to prevent enamel demineralization and promote remineralization of enamel subsurface lesions in animal and human in situ caries models. Improved casein phosphopeptide-amorphous calcium phosphate complexes (CPP-ACP) and CPP-stabilized amorphous calcium fluoride phosphate complexes (CPP-ACFP), including preferred complexes formed at a pH of 5 to 6.5, are also described in WO 2006 / 056013 and WO 2006 / 135982.

[0009] The CPPs active in the complex formation may or may not be part of the full-length casein protein. Examples of active CPPs that can be isolated after trypsin digestion of full-length casein are specified in U.S. Pat. No. 5,015,628, such as the peptide Bos α s1 -Casein X-5P (f59-79), Bos β-casein X-4P (f1-25), Bos α s2 -Casein X-4P (f46-70), and Bos α s2 -Casein X-4P (f1-21)

[0010] Although CPP-ACP and CPP-ACFP complexes are effective in remineralizing hypomineralized enamel, current manufacturing methods limit the amount of CPP-ACP or CPP-ACFP that can be used in liquid form due to the complex's tendency to crosslink and form a gel. Gel formation reduces the activity (bioavailability) of the ions required to remineralize enamel subsurface lesions. This is a clinically important limitation because remineralization is a slow process; several months may be required at a 10% CPP-ACP or CPP-ACFP concentration to achieve significant remineralization. Summary of the Invention [Problem to be solved by the invention]

[0011] There is a need to provide improved or alternative treatments for hypocalcified lesions.

[0012] The reference herein to any prior art is not an admission or implication that this prior art forms part of the common general knowledge in any jurisdiction, or that this prior art would be understood by, considered relevant, and / or could reasonably be inferred to be combined with other elements of prior art by a person skilled in the art. [Means for solving the problem]

[0013] In one aspect, the present invention provides a method for surface or subsurface mineralization of teeth, comprising the steps of: Simultaneously or subsequently with a step of contacting the surface or subsurface of the tooth with stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP), heating the surface or subsurface of the tooth to which the stabilized ACP and / or ACFP has been applied or has been applied to a temperature greater than 37°C; The present invention provides a method comprising:

[0014] In another aspect, the present invention provides a method for surface or subsurface mineralization of teeth, comprising the steps of: contacting the tooth surface or subsurface with a liquid composition comprising greater than 20% w / v stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); The present invention provides a method comprising:

[0015] In another aspect, the present invention provides a method for surface or subsurface mineralization of teeth, comprising the steps of: Simultaneously or subsequently with contacting the surface or subsurface of the tooth with a liquid composition comprising greater than 20% w / v stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP), heating the surface or subsurface of the tooth to which a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP has been applied or to which the liquid composition has been applied to a temperature greater than 37°C; The present invention provides a method comprising:

[0016] In a further aspect of the present invention, there is provided a method for remineralizing dental lesions, comprising the steps of: contacting the surface or subsurface of a hypomineralized tooth with a liquid composition having a pH of 5 or greater but not greater than 9, the liquid composition comprising at least 40% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); The method further comprises the step of: remineralizing the dental lesion. Preferably, the liquid composition has a pH of 6 or more but 8 or less, such as 7 or more but 8 or less.

[0017] In any aspect of the invention, the method includes heating the surface or subsurface of the tooth to which the stabilized ACP and / or ACFP (e.g., in a liquid composition) has been applied or has been applied to a temperature of 40°C or higher, 45°C or higher, 50°C or higher, 55°C or higher, 60°C or higher, or 65°C or higher.

[0018] In any aspect of the invention, the method includes heating the surface or subsurface of the tooth to which the stabilized ACP and / or ACFP (e.g., in a liquid composition) has been applied or has been applied to a temperature greater than 37°C but less than or equal to 65°C, greater than 40°C but less than or equal to 65°C, greater than 45°C but less than or equal to 65°C, greater than 50°C but less than or equal to 65°C, greater than 55°C but less than or equal to 65°C, or greater than 60°C but less than or equal to 65°C.

[0019] In any aspect of the present invention, the liquid composition comprising more than 20% w / v stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) includes 25% w / v or more, 30% w / v or more, 35% w / v or more, 40% w / v or more, 45% w / v or more, 50% w / v or more, 55% w / v or more, 60% w / v or more, 65% w / v or more stabilized ACP and / or ACFP, 70% w / v or more stabilized ACP and / or ACFP, or 75% w / v or more stabilized ACP and / or ACFP.

[0020] In any embodiment of the present invention, a liquid composition comprising more than 20% w / v stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) is , stabilized ACP and / or ACFP of more than 20% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 25% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 30% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 35% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 40% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 45% w / v but less than 80% w / v, stabilized ACP and / or ACFP of more than 50% w / v but less than 80% w / v, % w / v but less than 80% w / v of stabilized ACP and / or ACFP, greater than 55% w / v but less than 80% w / v of stabilized ACP and / or ACFP, greater than 60% w / v but less than 80% w / v of stabilized ACP and / or ACFP, greater than 65% w / v but less than 80% w / v of stabilized ACP and / or ACFP, greater than 70% w / v but less than 80% w / v of stabilized ACP and / or ACFP, or greater than 75% w / v but less than 80% w / v of stabilized ACP and / or ACFP.

[0021] In any embodiment of the present invention, the liquid composition comprises more than 40% w / w of phosphopeptide (PP) stabilized ACP and / or ACFP, more than 45% w / w, more than 50% w / w of stabilized ACP and / or ACFP, more than 55% w / w of stabilized ACP and / or ACFP, more than 60% w / w of stabilized ACP and / or ACFP, more than about 65% w / w of stabilized ACP and / or ACFP, more than about 70% w / w of stabilized ACP and / or ACFP, or more than about 75% w / w of stabilized ACP and / or ACFP.

[0022] In any embodiment of the invention, the liquid composition comprises more than 40% w / v phosphopeptide (PP) stabilized ACP and / or ACFP, more than 40% w / v but less than 80% w / v stabilized ACP and / or ACFP, more than 45% w / v but less than 80% w / v stabilized ACP and / or ACFP, more than 50% w / v but less than 80% w / v stabilized ACP and / or ACFP, more than 55% w / v stabilized ACP and / or ACFP but less than 80% w / v of stabilized ACP and / or ACFP, more than 60% w / v but less than 80% w / v of stabilized ACP and / or ACFP, more than 65% w / v but less than 80% w / v of stabilized ACP and / or ACFP, more than 70% w / v but less than 80% w / v of stabilized ACP and / or ACFP, or more than 75% w / v but less than 80% w / v of stabilized ACP and / or ACFP.

[0023] In either embodiment, the liquid composition is degassed. Degassing can be by any method that creates a negative pressure above the liquid composition. Exemplary methods include a vacuum pump or system, such as a venturi vacuum water system.

[0024] In any embodiment of the present invention, the stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) is phosphopeptide stabilized. Preferably, the phosphopeptide (defined below) is a casein phosphopeptide.

[0025] In either embodiment, the calcium ion content of the stabilized ACP or ACFP complex is greater than about 30 moles per mole of PP. Preferably, the calcium ion content is: It is in the range of about 30 to 100 moles of calcium per mole of PP. More preferably, the calcium ion content is in the range of about 30 to about 50 moles of calcium per mole of PP.

[0026] In a preferred embodiment of each aspect of the invention, the phosphopeptide-stabilized ACP or ACFP complex in the composition has tightly bound calcium and loosely bound calcium, the bound calcium in this complex being less than the tightly bound calcium in the ACP or ACFP complex formed at a pH of 7.0. Optionally, the ACP or ACFP is present primarily in its base form.

[0027] In either embodiment, the stabilized ACP complex is a stannous-associated phosphopeptide (PP) ACP complex, and the stabilized ACFP complex is a stannous-associated phosphopeptide (PP) stabilized amorphous calcium fluoride phosphate (ACFP) complex.

[0028] In either embodiment, the ACP and / or ACFP complexes are present in the form of casein phosphopeptide-stabilized ACP and / or ACFP complexes.

[0029] Preferably, the phase of the ACP is predominantly (i.e., >50%) basic, and the ACP is predominantly Ca. 2+ , PO4 3- , and OH - The basic phase of ACP has the general formula [Ca3(PO4)2] x It can be represented by the formula [Ca2(PO4)(OH)], where x≧1. Preferably, x=1 to 5. More preferably, x=1, i.e., the two components of the above formula are present in equal proportions. Thus, in one embodiment, the basic phase of ACP is represented by the formula Ca3(PO4)2Ca2(PO4)(OH).

[0030] Preferably, the phase of the ACFP is predominantly (i.e., >50%) basic, and the ACFP is predominantly Ca. 2+ , PO4 3- , and F- The basic phase of ACFP has the general formula [Ca3(PO4)2] x [Ca2(PO4)F] y where x≧1 when y=1, or y≧1 when x=1. Preferably, y=1 and x=1 to 3. More preferably, y=1 and x=1, i.e., the two components in the above formula are present in the same ratio. Thus, in one embodiment, the basic phase of ACFP is represented by the formula Ca3(PO4)2Ca2(PO4)F.

[0031] In one embodiment, the ACP complex consists essentially of phosphopeptide, calcium, phosphate, and hydroxide ions, and water. Preferably, the complex further comprises stannous ions.

[0032] In one embodiment, the ACFP complex consists essentially of phosphopeptide, calcium, phosphate, fluoride, and hydroxide ions, and water. Preferably, the complex further comprises stannous ions.

[0033] In one aspect, the present invention provides a method for treating fluorosis, comprising: simultaneously with or subsequent to contacting the fluorotic lesions, preferably in the dental enamel, with stabilized ACP and / or ACFP; heating the fluorotic lesion to which the stabilized ACP and / or ACFP has been applied or to which it has been applied to a temperature greater than 37°C; The present invention provides a method comprising:

[0034] In another aspect, the present invention provides a method for treating fluorosis, comprising: A method is provided which comprises contacting a fluorotic lesion, preferably in dental enamel, with a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP.

[0035] In another aspect, the present invention provides a method for treating fluorosis, comprising: Simultaneously or sequentially with contacting fluorotic lesions, preferably in dental enamel, with a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, heating the fluorotic lesion to a temperature above 37°C to which a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP has been applied or to which the liquid composition has been applied; The present invention provides a method comprising:

[0036] In one aspect, the present invention provides a method for treating dental caries, comprising: Simultaneously or subsequently to contacting the caries lesion with stabilized ACP and / or ACFP, heating the carious lesion to which the stabilized ACP and / or ACFP has been applied or to which it has been applied to a temperature greater than 37°C; The present invention provides a method comprising:

[0037] In another aspect, the present invention provides a method for treating dental caries, comprising: A method is provided that includes contacting a carious lesion with a liquid composition that includes greater than 20% w / v stabilized ACP and / or ACFP.

[0038] In another aspect, the present invention provides a method for treating dental caries, comprising: simultaneously with or subsequent to contacting the caries lesion with a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP; heating a carious lesion to which a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP has been or has been applied to a temperature greater than 37°C; The present invention provides a method comprising:

[0039] In any embodiment of the invention, the method, use, or composition of the invention can be used to treat dental erosion, wherein lesions in the dental enamel caused by dental erosion are contacted with stabilized ACP and / or ACFP, or a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP.

[0040] In any aspect of the invention, the method, use, or composition of the invention can be used to reduce white spot lesions, preferably on dental enamel, by contacting the white spot lesions with stabilized ACP and / or ACFP, or a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP.

[0041] In any aspect of the invention, the method, use, or composition of the invention can be used to remineralize lesions in dental enamel or dentin, in which the lesion, preferably on dental enamel, is contacted with stabilized ACP and / or ACFP, or a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP.

[0042] In any embodiment of the present invention, the liquid composition comprises about 40% w / v or more stabilized ACP.

[0043] In any embodiment of the present invention, the liquid composition comprises about 50% w / v or more stabilized ACFP.

[0044] In any embodiment of the invention, the surface or subsurface of the tooth, or lesion (such as caused by fluorosis, caries, white spots, or acid erosion), can be heated to a temperature of about 45° C. or higher. Preferably, this temperature does not exceed 65° C.

[0045] In any embodiment of the present invention, the pH of the liquid composition is 6 or less. Preferably, the pH is 5.5 or less. Alternatively, the pH of the liquid composition is 5 or more but 9 or less, preferably 6 or more but 8 or less, and most preferably 7 or more but 8 or less.

[0046] In one embodiment, when the % w / v of stabilized ACP and / or ACFP is greater than 20% w / v but less than 40% w / v, the pH of the liquid composition is between about 5 and about 8, preferably about 5 to about 7, preferably between 5 and 7. In another embodiment, when the % w / v of stabilized ACP and / or ACFP is greater than 40% w / v, the pH of the liquid composition is 6 or less. Preferably, the pH is 5.5 or less.

[0047] In any of the aspects of the invention described herein, the stabilized ACP or ACFP, a liquid composition comprising the stabilized ACP or ACFP, and / or heat is applied to the mouth, tooth, or lesion by a dental health care professional.

[0048] In any embodiment of the present invention, the surface or subsurface of the tooth, or lesion (eg, caused by fluorosis, caries, white spots, or acid erosion), can be heated for about 1 to 60 minutes, or about 1 to 30 minutes.

[0049] Preferably, the stabilized ACP and / or ACFP, or the liquid composition containing the stabilized ACP and / or ACFP, is contacted with the tooth surface for about 1 minute to 2 hours, or 5 minutes to 60 minutes, or about 10 minutes.

[0050] In any aspect, the tooth surface or subsurface or lesion is one in need of such treatment. Thus, the present invention includes, in addition to the steps of any of the methods described herein, the step of identifying a subject suffering from fluorosis, caries, dentin hypersensitivity, or tartar, white spot lesion; a fluorosis lesion; a caries lesion; or a lesion caused by dental erosion.

[0051] In another aspect, the present invention provides a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP. Preferably, the liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP comprises 25% w / v or more, 30% w / v or more, 35% w / v or more, 40% w / v or more, 45% w / v or more, 50% w / v or more, 55% w / v or more, 60% w / v or more, 65% w / v or more, 70% w / v or more, or 75% w / v or more stabilized ACP and / or ACFP.

[0052] In this aspect of the invention, the liquid composition may comprise more than 20% w / v stabilized ACP and / or ACFP but less than 80% w / v stabilized ACP and / or ACFP, more than 25% w / v stabilized ACP and / or ACFP but less than 80% w / v stabilized ACP and / or ACFP, more than 30% w / v stabilized ACP and / or ACFP but less than 80% w / v stabilized ACP and / or ACFP, more than 35% w / v stabilized ACP and / or ACFP but less than 80% w / v stabilized ACP and / or ACFP, 40% stabilized ACP and / or ACFP greater than 45% w / v but less than 80% w / v; stabilized ACP and / or ACFP greater than 50% w / v but less than 80% w / v; stabilized ACP and / or ACFP greater than 55% w / v but less than 80% w / v; stabilized ACP and / or ACFP greater than 60% w / v. The stabilized ACP and / or ACFP may comprise more than 80% w / v of stabilized ACP and / or ACFP but less than 80% w / v, more than 65% w / v of stabilized ACP and / or ACFP but less than 80% w / v, more than 70% w / v of stabilized ACP and / or ACFP but less than 80% w / v, or more than 75% w / v of stabilized ACP and / or ACFP but less than 80% w / v.

[0053] In this embodiment, the liquid composition comprises more than 40% w / w phosphopeptide (PP) stabilized ACP and / or ACFP, more than 45% w / w, more than 50% w / w stabilized ACP and / or ACFP, more than 55% w / w stabilized ACP and / or ACFP, more than 60% w / w stabilized ACP and / or ACFP, more than about 65% w / w stabilized ACP and / or ACFP, more than about 70% w / w stabilized ACP and / or ACFP, or more than about 75% w / w stabilized ACP and / or ACFP.

[0054] In this embodiment, the liquid composition comprises more than 40% w / w of phosphopeptide (PP) stabilized ACP and / or ACFP, more than 40% w / w of stabilized ACP and / or ACFP but less than 80% w / w of stabilized ACP and / or ACFP, more than 45% w / w of stabilized ACP and / or ACFP but less than 80% w / w of stabilized ACP and / or ACFP, more than 50% w / w of stabilized ACP and / or ACFP but less than 80% w / w of stabilized ACP and / or ACFP, more than 55% w / w of stabilized ACP and / or ACFP but less than 80% w / w of stabilized ACP and / or ACFP, The stabilized ACP and / or ACFP may be less than 0% w / w, more than 60% w / w but less than 80% w / w, more than 65% w / w but less than 80% w / w, more than 70% w / w but less than 80% w / w, or more than 75% w / w but less than 80% w / w.

[0055] In any of the aspects described herein, the liquid composition can further include fluoride ions, preferably free fluoride ions. The fluoride ions can be present in the liquid composition at a concentration ranging from about 200 ppm to 50,000 ppm. In a preferred embodiment, the fluoride ions are present at a concentration ranging from about 2,600 ppm to about 10,000 ppm. In a more preferred embodiment, the fluoride ions are present in the liquid composition at a concentration of about 8,200 ppm or about 6,500 ppm. The fluoride ions can be present in the liquid composition at any ppm level described herein, particularly in the Examples. In any of the embodiments, the fluoride ions are present at a concentration of about 2,600 ppm, 3,900 ppm, 5,200 ppm, 6,500 ppm, or 7,800 ppm. Typically, fluoride ions are present at a concentration of about 2600 ppm for 20% w / v stabilized ACP or ACFP, about 3,260 ppm for 25% w / v stabilized ACP or ACFP, about 3,900 ppm for 30% w / v stabilized ACP or ACFP, about 4,890 ppm for 38% stabilized ACP or ACFP, 5,200 ppm for 40% w / v stabilized ACP or ACFP, 6,500 ppm for 50% w / v stabilized ACP or ACFP, about 8,200 ppm for 63% stabilized ACP or ACFP, and about 9,900 ppm for 75% stabilized ACP or ACFP. In another embodiment, fluoride ions are present at a concentration of about 5,200 ppm for 40% w / w CPP-ACP or about 7,800 ppm for 60% w / w CPP-ACP. Preferably, the stabilized ACP and / or ACFP is phosphopeptide-stabilized. Preferably, the phosphopeptide is a casein phosphopeptide.

[0056] Any of the compositions described herein can be used in any one of the methods described herein, provided that the composition is a physiologically acceptable composition as described herein.

[0057] In another aspect, the present invention provides: Calcification of the surface or subsurface of a tooth, or any of the lesions described herein; or Treatment or prevention of one or more of each of dental caries, dental caries, dental erosion, white spot lesions, and fluorosis; The present invention provides a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP for use in

[0058] In a further aspect, there is provided use of a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP in the manufacture of a composition or medicament for the treatment and / or prevention of one or more of tooth surface or subsurface mineralization, or dental caries, dental caries, dental erosion, and fluorosis.

[0059] In a further aspect, there is provided the use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) in the manufacture of a product comprising or consisting of a liquid composition for surface or subsurface remineralization of teeth, comprising: Use is provided wherein the liquid composition comprises at least 40% w / w of said phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) and has a pH of not less than pH 5 but not more than pH 9. In one embodiment, the product is a cosmetic.

[0060] The present invention also relates to a kit for treating or preventing one or more of dental caries, fluorosis, and dental erosion, or mineralization of dental surfaces or subsurfaces or lesions, as described herein, comprising: (a) a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP; and (b) a heat source. Desirably, the kit further comprises instructions for use in any of the methods or uses described herein. The instructions may describe the use of the kit for the treatment or prevention of one or more of dental caries, dental caries, dental erosion, and fluorosis, respectively. In one embodiment, the liquid composition is present in an amount appropriate for treating a patient. Preferably, the stabilized ACP and / or ACFP is phosphopeptide (PP)-stabilized. Preferably, the phosphopeptide (defined below) is a casein phosphopeptide. Preferably, the ACP or ACFP is in the form of a casein phosphopeptide-stabilized ACP or ACFP complex.

[0061] The composition or kit of the present invention can further comprise a fluoride ion source. The fluoride ion can be derived from any suitable source. The fluoride ion source can include free fluoride ions or fluoride salts. Examples of fluoride ion sources include, but are not limited to, sodium fluoride, sodium monofluorophosphate, stannous fluoride, sodium silicofluoride, silver fluoride, amine fluoride, or any metal ion fluoride salt. The fluoride ion source can be hypofluorite. These fluoride ion sources can be provided in a solution (typically an aqueous solution) or suspension.

[0062] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, comprising: mixing a solvent with a powder comprising or consisting of stabilized ACP and / or ACFP; maintaining a pH below 7; Preferably, the pH is maintained at or below 6, preferably the pH is maintained at or below 5.5.

[0063] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, comprising: mixing a solvent with a powder comprising or consisting of stabilized ACP and / or ACFP; reducing the pH to less than 7; Preferably, the pH is lowered to below 6, preferably below 5.5. Typically, the pH is maintained below 7, more preferably the pH is maintained at below 6, even more preferably below 5.5.

[0064] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, comprising: mixing a fluoride-containing solution with a powder comprising or consisting of stabilized ACP and / or ACFP; adjusting the pH to between 6.5 and 8; Preferably, the pH is adjusted to about pH 7.8. Preferably, the pH is adjusted using HCl.

[0065] In this embodiment, the method further comprises, after adjusting the pH, mixing the solution for at least about 10 minutes, at least about 20 minutes, or at least about 30 minutes.

[0066] In this embodiment, the method further comprises the step of degassing the liquid composition.

[0067] In either embodiment, the step of mixing the solvent and the powder comprising or consisting of the PP-stabilized ACP and / or ACFP comprises adding the solvent to the powder, or alternatively, this step comprises adding the powder to the solvent.

[0068] In any of the methods or processes for preparing a liquid composition described herein, the method or process further comprises the step of degassing the liquid composition. Degassing can be accomplished by any method that creates a negative pressure above the liquid composition, such as the methods described herein.

[0069] In any of the methods or processes for preparing a liquid composition described herein, the method or process further comprises the step of mixing the liquid composition with a solution comprising fluoride ions.

[0070] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, comprising or consisting of the steps described in Example 2 or 4 herein.

[0071] In any embodiment, the present invention provides a method for preparing a powder comprising or consisting of stabilized ACP and / or ACFP by: mixing one or more solutions comprising phosphopeptides, calcium ions, phosphate ions, hydroxide ions, and optionally fluoride ions, while maintaining a pH above about 7.0, preferably about 9, to form a solution comprising stabilized ACP and / or ACFP; Drying the solution containing stabilized ACP and / or ACFP; thereby forming a powder comprising or consisting of stabilized ACP and / or ACFP; Preferably, the drying is spray drying or freeze drying.

[0072] In one embodiment, the method or process comprises: The method further includes filtering the solution containing the stabilized ACP and / or ACFP prior to drying to form a retentate, which is then dried to form a powder containing or consisting of the stabilized ACP and / or ACFP.

[0073] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising at least 40% w / w of PP-stabilized ACP and / or ACFP, comprising: mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP; reducing the pH to less than 9, preferably less than 8; Preferably, the solvent comprises a fluoride.

[0074] In this embodiment, the method further comprises the step of stirring the liquid composition after lowering the pH, preferably for at least 5, 10, 15, 20, 25, or 30 minutes.

[0075] In this embodiment, the liquid composition is degassed, preferably by placing the solution under vacuum, most preferably for 24 hours, to remove any trapped air bubbles.

[0076] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising at least 40% w / w, preferably 60% w / w, of a PP-stabilized ACP and / or ACFP, comprising or consisting of the steps described in Example 4 herein.

[0077] In any method or process for preparing a liquid composition comprising more than 20% w / v or more than 40% w / w stabilized ACP and / or ACFP, the solvent is water.

[0078] In any method or process for preparing a liquid composition containing more than 20% w / v stabilized ACP and / or ACFP, the pH is reduced or maintained using 1-10 M HCl, or 11 M HCl.

[0079] In any embodiment, the method or process for preparing a liquid composition comprising more than 20% w / v stabilized ACP and / or ACFP may be for preparing a liquid composition comprising 25% w / v or more, 30% w / v or more, 35% w / v or more, 40% w / v or more, 45% w / v or more, 50% w / v or more, 55% w / v or more, 60% w / v or more, 65% w / v or more, 70% w / v or more, 75% w / v or more, or 80% w / v or more stabilized ACP and / or ACFP.

[0080] In any method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, the stabilized ACP or ACFP is a CPP-ACP or CPP-ACFP described herein.

[0081] In any method or process for preparing a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP, the liquid composition is for use in any dental treatment method, preferably a method described herein (e.g., surface or subsurface mineralization of teeth).

[0082] In another aspect, the present invention provides a liquid composition comprising greater than 20% w / v stabilized ACP and / or ACFP prepared by a method or process described herein.

[0083] Further aspects of the invention, and further embodiments of the aspects described in the preceding paragraphs, will become apparent from the following description, given by way of example and referring to the accompanying drawings, in which: [Brief explanation of the drawings]

[0084] [Figure 1]Effect of temperature on remineralization of enamel subsurface lesions in vitro. [Figure 2] Remineralization by high concentrations of CPP-ACP in the presence of fluoride. [Figure 3] Remineralization with high concentrations of CPP-ACP. DETAILED DESCRIPTION OF THE INVENTION

[0085] It will be understood that the invention disclosed and defined herein extends to any and all alternative combinations of two or more of the individual features mentioned or apparent from the text or drawings, all of which constitute various alternative aspects of the invention.

[0086] Further aspects of the present invention and further embodiments of the aspects described in the above paragraphs will become apparent from the following description, given by way of example only, and with reference to the accompanying drawings, in which:

[0087] Reference will now be made in more detail to specific embodiments of the invention. While the invention will be described in conjunction with the embodiments, it will be understood that it is not intended that the invention be limited to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present invention as defined by the claims.

[0088] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0089] All patents and publications cited herein are incorporated by reference in their entirety.

[0090] For the purposes of describing this specification, terms used in the singular will also include the plural and vice versa.

[0091] As used herein, unless the context requires otherwise, the term "comprise" and variations of that term, such as "comprising," "comprises," and "comprised," are not intended to exclude additional additives, ingredients, integers, and steps. As used herein, unless the context requires otherwise, "comprise" and "include" can be used interchangeably.

[0092] One aspect of the invention described herein is based on the surprising discovery that compositions with high concentrations of stabilized ACP and / or ACFP are achievable that remain in a liquid state (i.e., do not form a gel). Prior to the present invention, it was believed that high concentrations of phosphopeptide-stabilized ACP and / or ACFP would result in compositions that formed gels or pastes, and all liquid compositions described to date have had relatively low concentrations of phosphopeptide-stabilized ACP and / or ACFP. The unexpected property of high-concentration compositions of phosphopeptide-stabilized ACP and / or ACFP remaining in a liquid state prior to application to the tooth surface or subsurface allows for more rapid penetration into hypomineralized areas. Without being bound by any theory or mechanism of action, higher concentrations of stabilized ACP and / or ACF It is believed that P is able to reach the lesion, thereby allowing for more rapid remineralization to a greater extent. This offers the advantage that highly concentrated liquid compositions can be applied directly onto the lesion by a dental professional, allowing for more extensive remineralization compared to the home application of less concentrated compositions such as pastes or mousses.

[0093] Yet another aspect of the invention described herein is based on the surprising discovery that applying surface or subsurface heating to the teeth, either simultaneously with or subsequent to application of stabilized ACP and / or ACFP, increases the extent of mineralization, even at relatively high temperatures.

[0094] Finally, the combination of a highly concentrated liquid composition and heating the teeth to which the stabilized ACP and / or ACFP has been or will be applied results in widespread and rapid remineralization.

[0095] Any heat source can be used in the method or use of the present invention to heat the surface or subsurface of the tooth. Heat sources suitable for use in light or radiation extraction and dental applications are well known in the art. Specific examples include dental curing lights, such as the X-Cure by Guilin Woodpecker Medical Instrument Co., Ltd., a 10W high-power blue LED.

[0096] As used herein, % w / v can be interpreted as equivalent to g / 100 ml.

[0097] As used herein, "stabilized ACP or ACFP" and "stabilized ACP or ACFP complex" are used interchangeably.

[0098] A stabilized ACP or ACFP complex as described herein may be the "closed" complex shown in Figure 2 of Cross et al., 2007.

[0099] Stabilized ACPs or ACFPs as referred to herein include stabilized ACPs or ACFPs as described in WO 2006 / 056013 (PCT / AU2005 / 001781), the contents of which are incorporated by reference.

[0100] In a preferred embodiment, the phosphopeptide-stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex has tightly bound and loosely bound calcium, the bound calcium in this complex being less than the tightly bound calcium in the ACP or ACFP complex formed at pH 7.0. Optionally, the ACP or ACFP is predominantly in the base form.

[0101] Stabilized ACP or ACFP complexes as referred to herein include stabilized ACP or ACFP complexes formed at a pH of less than 7.0. Preferably, the complexes are formed at a pH within the range of about 5.0 to less than 7.0. More preferably, the complexes are formed at a pH range of about 5.0 to about 6.0. In a preferred embodiment, the complexes are formed at a pH of about 5.5. Preferably, the ACP or ACFP in the complex is predominantly in the base form.

[0102] Stabilized ACP: (i) obtaining a solution containing at least one phosphopeptide; (ii) mixing a solution containing calcium ions, phosphate ions, and hydroxide ions while maintaining a pH between about 5.5 and 9; It can be produced by a method comprising:

[0103] In one embodiment, the pH is maintained at or below about 7.0.

[0104] Stabilized ACFP is: (i) obtaining a solution containing at least one phosphopeptide; (ii) mixing a solution containing calcium ions, phosphate ions, hydroxide ions, and fluoride ions while maintaining a pH between about 5.5 and 9; It can be produced by a method comprising:

[0105] In one embodiment, the pH is maintained at or below about 7.0.

[0106] A phosphopeptide-stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex may be formed in which the ACP in the complex has both tightly bound calcium and loosely bound calcium, or in which the tightly bound calcium in the complex is less than the tightly bound calcium in the ACP or ACFP complex formed at pH 7.0, and the ACP or ACFP is primarily in the base form: a) mixing a first solution containing calcium ions, a second solution containing phosphate ions, and optionally a third solution containing fluoride ions with a solution containing a phosphopeptide and a solvent at a pH of about 5 to less than 7; b) maintaining the pH of the solution during mixing at about 5.0 to less than 7.0 by adding hydroxide ions; It can include cases where it can be obtained or obtained by.

[0107] "Tightly" and "loosely" bound calcium and phosphate in ACP or ACFP can be measured using analytical ultrafiltration. Briefly, a solution of mixed phosphopeptide, calcium, phosphate, and optionally fluoride, while maintaining a pH of about 7.0 or less, can be first filtered through a 0.1 micron filter to remove free calcium and phosphate that are not associated with complexes. This free calcium and phosphate resides in the filtrate and is discarded. Any free calcium or phosphate that is not associated with complexes is not bioavailable, i.e., is not delivered to the teeth by the phosphopeptide. The retentate resulting from 0.1 micron filtration can be further analyzed by centrifugation at 1,000 g for 15 minutes through a 3,000 mw cutoff filter. The resulting filtrate contains calcium and phosphate that are loosely bound or associated with complexes. At this centrifugal force, calcium and phosphate that are not tightly bound to complexes are released and move into the filtrate. The tightly bound Ca and Pi in complexes are retained in the retentate. The amount of tightly bound Ca and Pi in the retentate can then be determined by subtracting the amount of Ca and Pi in the filtrate from the total amount of Ca and Pi in the retentate from the 0.1 micron filtration.

[0108] Stabilized ACP or ACFP complexes as referred to herein include stabilized ACP or ACFP complexes as described in International Publication No. WO 2006 / 135982 (PCT / AU2006 / 000885), the contents of which are incorporated by reference.

[0109] "Superloaded" phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complexes. The complexes can be formed at any pH (e.g., 3-10). Preferably, the phosphopeptide contains the sequence -ABC-, where A is a phosphoamino acid, preferably phosphoserine, B is any amino acid, such as a phosphoamino acid, and C is glutamic acid, aspartic acid, or a phosphoamino acid. The phosphoamino acid may be phosphoserine. The PP is superloaded with calcium and phosphate ions. The sodium ions may be in the range of 30 to 1000 moles of Ca per mole of PP, or in the range of 30 to 100 or 30 to 50 moles of Ca per mole of PP. In another embodiment, the number of moles of Ca per mole of PP is at least 25, 30, 35, 40, 45, or 50.

[0110] The phosphopeptide- or phosphoprotein-(PP)-stabilized amorphous calcium phosphate or amorphous calcium fluoride phosphate complex may have a calcium ion content greater than about 30 moles of calcium per mole of PP. In a preferred embodiment, the calcium ion content is in the range of about 30 to 100 moles of calcium per mole of PP. More preferably, the calcium ion content is in the range of about 30 to about 50 moles of calcium per mole of PP.

[0111] Phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complexes (i) obtaining a solution containing calcium, inorganic phosphate, and fluoride (optional); (ii) mixing (i) with a solution containing PP-ACP; It can be produced by a method comprising:

[0112] In one preferred embodiment, the PP is a casein phosphopeptide (CPP).

[0113] The PP-stabilized ACP and / or ACFP complex can further comprise at least the same amount of calcium phosphate by weight. Preferably, the calcium phosphate is CaHPO4. Preferably, the calcium phosphate (e.g., CaHPO4) is dry-blended with the PP-stabilized ACP and / or ACFP complex. In a preferred embodiment, the ratio of PP-ACP and / or PP-ACFP complex to calcium phosphate is about 1:1-50, more preferably about 1:1-25, and more preferably about 1:5-15. In one embodiment, the ratio of PP-ACP and / or PP-ACFP complex to calcium phosphate is about 1:10.

[0114] Oral care formulations containing phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) complexes having a calcium ion content greater than about 30 moles of calcium per mole of PP when used in the oral cavity: (i) obtaining a powder comprising a PP-ACP and / or PP-ACFP complex; (ii) dry blending with an effective amount of calcium phosphate; (iii) formulating an oral care formulation from the dry-blended PP-ACP and / or PP-ACFP and calcium phosphate mixture; It can be produced by a method comprising:

[0115] Preferably, the form of calcium phosphate for dry blending is any soluble calcium phosphate, including but not limited to CaHPO4, Ca2HPO4, and calcium lactate.

[0116] The compositions described herein can further comprise free fluoride ions. Fluoride ions can be derived from any suitable source. Fluoride ion sources can include free fluoride ions or fluoride salts. Examples of fluoride ion sources include, but are not limited to, sodium fluoride, sodium monofluorophosphate, stannous fluoride, sodium silicofluoride, and amine fluorides. These can be provided as a solution (typically an aqueous solution) or a suspension.

[0117] Fluoride ions are preferably present in the composition in an amount greater than 1 ppm. More preferably, the amount is greater than 3 ppm. In another embodiment, the amount is preferably greater than 10 ppm. In the exemplary embodiment described below, the amount can be hundreds or thousands of ppm. Typically, free fluoride ions are in the range of 1000 ppm to 50,000 ppm F. ppm F can be any of the amounts or concentrations described herein. Fluoride content is typically measured as ppm in oral compositions using methods commonly used in the art. When fluoride is obtained from a source with stabilized ACP, ppm refers to the fluoride concentration in the source, which is typically a solution or suspension of bioavailable fluoride.

[0118] The tin-associated ACP or ACFP complexes described herein include any of those described in International Publication No. PCT / AU2014 / 050447, the contents of which are incorporated by reference in their entirety.

[0119] The compositions described herein for use in the methods of use of the invention can include a stannous-associated ACP or ACFP complex. The compositions can include 2% CPP-ACP and 290 ppm fluoride, including 220 ppm as stannous fluoride and 70 ppm as sodium fluoride.

[0120] In the context of the present description, a "phosphopeptide" refers to an amino acid sequence in which at least one amino acid is phosphorylated. Preferably, a phosphopeptide comprises one or more amino acid sequences -ABC-, where A is a phosphoamino residue, B is any aminoacyl residue, including phosphoamino residues, and C is selected from glutamyl, aspartyl, or phosphoamino residues. Any phosphoamino residue may independently be a phosphoseryl residue. B is desirably a residue whose side chain is neither relatively large nor hydrophobic. It may be Gly, Ala, Val, Met, Leu, Ile, Ser, Thr, Cys, Asp, Glu, Asn, Gln, or Lys.

[0121] In another embodiment, at least two phosphoamino acids in this sequence are preferably adjacent. Preferably, the phosphopeptide comprises the sequence ABCDE, where A, B, C, D, and E are independently phosphoserine, phosphothreonine, phosphotyrosine, phosphohistidine, glutamic acid, or aspartic acid, and at least two, preferably three, of A, B, C, D, and E are phosphoamino acids. In a preferred embodiment, the phosphoamino acid residues are phosphoserine, most preferably three consecutive phosphoserine residues. It is also preferred that D and E are independently glutamic acid or aspartic acid.

[0122] In one embodiment, the ACP or ACFP is stabilized by casein phosphopeptides (CPPs), which are in the form of intact casein or fragments of casein, and the complex formed preferably has the formula [CPP(ACP) 8]n or [(CPP)(ACFP)8]n, where n is 1 or greater, e.g., 6. The complex formed may be a colloidal complex, the core particle of which aggregates to form large (e.g., 100 nm) colloidal particles in water. Thus, PP may be a casein protein or a phosphopeptide.

[0123] PP may be obtained from any source and may exist in the context of a larger polypeptide, including full-length casein polypeptides, and may be prepared by trypsin or other enzymatic or chemical digestion of casein or another phosphoamino acid-rich protein, such as phosphitin, or by chemical or recombinant synthesis. The nucleic acid fragment may be isolated by the above-mentioned method, provided that it contains the aforementioned sequence -ABC- or ABCDE. The sequence flanking this core sequence may be any sequence. However, s1 (59 79), β(1-25), α s2 (46-70), and α s2 The flanking sequences in (1-21) are preferred. The flanking sequences may optionally be modified by deletion, addition, or conservative substitution of one or more residues. The amino acid composition and sequence of the flanking region are not critical.

[0124] Examples of conservative substitutions are shown in Table A below.

[0125] [Table 1]

[0126] The flanking sequences may also include residues of unnatural amino acids. Commonly encountered amino acids that are not encoded by the genetic code include: 2-aminoadipic acid (Aad) in the case of Glu and Asp; 2-aminopimelic acid (Apm) in the case of Glu and Asp; 2-aminobutyric (Abu) acid in the case of Met, Leu, and another aliphatic amino acid; 2-aminoheptanoic acid (Ahe) in the case of Met, Leu, and another aliphatic amino acid; 2-aminoisobutyric acid (Aib) in the case of Gly; cyclohexylalanine (Cha) in the case of Val and Leu and Ile; homoarginine (Har) in the case of Arg and Lys; 2,3-diaminopropionic acid (Dpr) in the case of Lys, Arg, and His; N-ethylglycine (EtGly) for Gly, Pro, and Ala; Asn, and N-ethylasparigine (EtAsn) in the case of Gln; hydroxylysine (Hyl) in the case of Lys; allohydroxylysine (AHyl) in the case of Lys; 3-(and 4) hydroxyproline (3Hyp, 4Hyp) in the cases of Pro, Ser, and Thr; Alloisoleucine (Alle) in the cases of Ile, Leu, and Val; ρ-amidinophenylalanine in the case of Ala; N-methylglycine (MeGly, sarcosine) in the cases of Gly, Pro, and Ala. N-methylisoleucine (MeIle) in the case of Ile; norvaline (Nva) in the case of Met and another aliphatic amino acid; norleucine (Nle) in the case of Met and another aliphatic amino acid; Ornithine (Orn) for Lys, Arg, and His; citrulline (Cit) and methionine sulfoxide (MSO) in the case of Thr, Asn, and Gln; In the case of Phe, N-methylphenylalanine (MePhe), trimethylphenylalanine, halo (F, Cl, Br and I) phenylalanine, triflourylphenylalanine.

[0127] In one embodiment, PP is α s1 (59-79)[1], β(1-25)[2], α s2 (46-70)[3], and α s2 (1-21)[4] is one or more phosphopeptides selected from the group consisting of: [1] Gln 59-Met-Glu-Ala-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ile-Val-Pro-Asn-Ser(P)-Val-Glu-Gln-Lys 79 (SEQ ID NO: 1) α s1 (59-79) [2] Arg 1 -Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-Ser(P)-Leu-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ser-Ile-Thr-Arg 25 (SEQ ID NO: 2) β(1-25) [3] Asn 46 -Ala-Asn-Glu-Glu-Glu-Tyr-Ser-Ile-Gly-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ser(P)-Ala-Glu-Val-Ala-Thr-Glu-Glu-Val-Lys 70 (SEQ ID NO: 3) α s2 (46-70) [4] Lys 1 -Asn-Thr-Met-Glu-His-Val-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ser-Ile-Ile-Ser(P)-Gln-Glu-Thr-Tyr-Lys 21 (SEQ ID NO: 4) α s2 (1-21).

[0128] In some preferred embodiments of the present invention, the liquid composition may be a mouthwash, rinse, spray, or the like. In such preparations, the vehicle is typically a water-alcohol mixture, preferably containing a humectant, as described below. Generally, the weight ratio of water to alcohol is in the range of about 1:1 to about 20:1. The total amount of the water-alcohol mixture in this type of preparation is typically in the range of about 70 to about 99.9% by weight of the preparation. The alcohol is typically ethanol or isopropanol. Ethanol is preferred.

[0129] It will be appreciated that, as is conventional, oral preparations will typically be sold or otherwise distributed in suitable labeled packaging. Thus, a bottle of mouthrinse will have labeling that substantially identifies it as a mouthrinse or mouthwash and describes how to use it.

[0130] Organic surfactants can be used in the compositions of the present invention to increase the preventive effect, promote thorough and complete dispersion of the active substance throughout the oral cavity, and make the composition more cosmetically acceptable. The organic surfactant material is preferably anionic, nonionic, or amphoteric, and preferably does not interact with the active substance. It is preferred to use a cleansing material as the surfactant, which imparts cleansing and foaming properties to the composition. Suitable examples of anionic surfactants include water-soluble salts of higher fatty acid monoglyceride monosulfates, such as the sodium salt of monosulfated monoglyceride of hydrogenated coconut oil fatty acid, higher alkyl sulfates, such as sodium lauryl sulfate, alkylaryl sulfates, such as sodium dodecylbenzene sulfate, higher alkyl sulfoacetates, 1,2-dihydroxypropane sulfonates, and the like. Examples of suitable amides include higher fatty acid esters of sarconite and substantially saturated higher aliphatic acyl amides of lower aliphatic aminocarboxylic acids, such as those having 12 to 16 carbon atoms in the fatty acid, alkyl group, or acyl group. Examples of the latter amides include N-lauroyl sarcosine and the sodium, potassium, and ethanolamine salts of N-lauroyl, N-myristoyl, or N-palmitoyl sarcosine, which should be substantially free of soap and similar higher fatty acid materials. The use of these sarconite compounds in the oral compositions of the present invention is particularly advantageous because these materials exhibit long-term, significant effects in inhibiting acid formation in the oral cavity by decomposing carbohydrates in addition to reducing the solubility of tooth enamel in acid solutions to some extent. Examples of water-soluble nonionic surfactants suitable for use are the condensation products of ethylene oxide with various reactive hydrogen-containing compounds having long hydrophobic chains (e.g., aliphatic chains of about 12 to 20 carbon atoms) that are reactive therewith; these condensation products ("ethoxamers") contain hydrophilic polyoxyethylene moieties, such as the condensation products of poly(ethylene oxide) with fatty acids, fatty alcohols, fatty amides, polyhydric alcohols (e.g., sorbitan monostearate), and polypropylene oxide (e.g., the Pluronic materials).

[0131] The surfactant is typically present in an amount of about 0.1 to 5% by weight. It should be noted that the surfactant may facilitate dissolution of the active agent of the present invention, thereby reducing the amount of solubilizing wetting agent required.

[0132] Various other materials can be incorporated into the oral preparations of the present invention, such as whitening agents, preservatives, silicones, chlorophyll compounds, and / or ammoniated materials such as urea, diammonium phosphate, and mixtures thereof. If present, these adjuvants are incorporated into the preparations in amounts that do not substantially adversely affect the desired properties and characteristics.

[0133] Any suitable flavoring or sweetening material may be used. Examples of suitable flavoring ingredients include flavor oils such as spearmint oil, peppermint oil, wintergreen oil, sassafras oil, clove oil, sage oil, eucalyptus oil, marjoram oil, cinnamon oil, lemon oil, and orange oil, and methyl salicylate. Suitable sweetening agents include sucrose, lactose, maltose, sorbitol, xylitol, sodium cyclamate, perillartine, AMP (aspartylphenylalanine methyl ester), saccharin, and the like. Preferably, the flavoring and sweetening agents, individually or combined, comprise from about 0.1% to more than 5% of the formulation.

[0134] In another embodiment, the compositions of the present invention described herein do not include a phosphate buffer and / or a calcium chelator. For example, any of the dentifrices described herein may be free of a phosphate buffer and / or a calcium chelator.

[0135] In one embodiment of the present invention, there is provided a composition wherein the composition does not include a phosphate buffer and / or a calcium chelator.

[0136] In another embodiment, the inventive compositions described herein are free of viscosity modifiers or contain 0.5-50% of viscosity modifiers.

[0137] In another embodiment, the compositions of the invention described herein do not contain sodium carboxymethylcellulose or do not contain 0.01 to 10% sodium carboxymethylcellulose having a degree of esterification of 0.7 to 1.0.

[0138] In one embodiment, the active ingredient of the composition consists essentially of a stabilized ACP or ACFP complex. become.

[0139] Although this specification refers specifically to human use, it will be clearly understood that the invention is also useful for veterinary purposes, and thus in all aspects the invention is useful for farm animals such as cattle, sheep, horses, and poultry, companion animals such as cats and dogs, and zoo animals.

[0140] The present invention also provides a kit comprising stabilized amorphous calcium phosphate (ACP) and / or stabilized amorphous calcium fluoride phosphate (ACFP), the kit being adapted for use in the aforementioned methods.

[0141] The present invention also provides kits comprising the liquid compositions described herein.

[0142] In any embodiment, the kit may further comprise a label or package insert with instructions for use in any of the methods described herein.

[0143] This kit includes: - a container containing a composition comprising stabilized amorphous calcium phosphate (ACP) and / or stabilized amorphous calcium fluoride phosphate (ACFP); - labelling or package insert with instructions for use, may include:

[0144] In certain embodiments, the kit can include one or more additional active ingredients or components for the treatment or prevention of a disease or condition described herein.

[0145] The kit can include a container and labeling or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, blister packs, and the like. The container can be formed from a variety of materials, such as glass or plastic. The container holds a therapeutic composition effective for treating a medical condition and can have a sterile access port (e.g., the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The labeling or package insert indicates that the therapeutic composition is used to treat a medical condition of choice. In one embodiment, the labeling or package insert includes instructions for use, indicating that the therapeutic composition can be used to treat a particular medical condition.

[0146] The kit can include (a) a liquid composition described herein; and (b) a second container containing a second active ingredient or component. The kit of this embodiment of the invention can further include a package insert indicating that the composition and other active ingredients can be used to treat the conditions described herein. Alternatively, or in addition, the kit can further include a second (or third) container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It can further include additional materials desirable from a commercial and user standpoint, such as other buffers, diluents, filters, needles, and syringes.

[0147] It is to be understood that the invention disclosed and defined herein extends to any and all alternative combinations of two or more of the individual features shown in or apparent from the text or drawings, all of which constitute various alternative aspects of the invention.

[0148] The invention will now be further described with reference to the following non-limiting examples. [Example]

[0149] Example 1 Effect of temperature on remineralization of enamel subsurface lesions. The purpose of these experiments was to investigate the effect of temperature on remineralization of enamel subsurface lesions using an in vitro model.

[0150] Solutions were prepared with CPP-ACP, and optionally NaF, to give 1.0% w / v CPP-ACP pH 5.5 or 1.0% w / v CPP-ACFP pH 5.5.

[0151] Tests were conducted at five different temperatures: (i) 25°C, (ii) 35°C, (iii) 45°C, (iv) 55°C, and (v) 65°C.

[0152] Using the method of Reynolds (J. Dent. Res. 1997, 76(9):1587-95), demineralized subsurface lesions of human dental enamel were created in enamel blocks of third molars.

[0153] Half of the blocks were kept as controls, and the other half of the blocks were remineralized by individually suspending them in 1.0% CPP-ACP + 725 ppm F for 14 days at five different temperatures (25, 35, 45, 55, and 65°C).

[0154] After remineralization, the enamel blocks were mounted, sectioned, and subjected to transverse microradiography and densitometric image analysis to determine the percent increase in mineral content (% remineralization), as previously described by Reynolds (1997, J Dent Res, supra). Figure 1 shows a direct correlation between increasing temperature and increasing amounts of remineralization.

[0155] [Table 2]

[0156] Example 2 A method for preparing a highly concentrated liquid composition containing CPP-ACP or CPP-ACP and having free fluoride is described below.

[0157] Just before precipitation or gelation (usually about 78 mM Ca 2+ A stock solution of 3.25 M CaCl2 and 1.25 M NaH2PO4 (pH 5.5) was added in approximately 30 aliquots to a 10-15% w / v tryptic casein digest (resulting in a final concentration of 48-76 mM inorganic phosphate). The solutions were added slowly (i.e., at less than approximately 1% volume per minute) with thorough mixing. One aliquot of the phosphate solution was added first, followed by one aliquot of the calcium solution. The pH of the bulk solution was maintained at 9.0 with thorough mixing using 1-10 M NaOH. Sodium hydroxide solution was added automatically by a pH stat, typically by adding hydroxide ion after each addition of calcium ion. After the addition of calcium ions, phosphate ions, and hydroxide ions was complete, the solution was filtered through a 0.1 micron filter and concentrated 1-2 times. The retentate was then washed with 1-2 volumes of water to remove salts and inactive (and bitter-tasting) peptides. The prepared CPP-ACP solution was then spray-dried or freeze-dried to obtain a white powder. This dried powder was then added to water and 1-10 M HCl was added to obtain a 20%-75% w / v CPP-ACP solution at pH 5.5, or NaF Addition of 25% w / v resulted in 3260 ppm F, 38% in 4890 ppm F, 50% in 6520 ppm F, 63% in 8151 ppm F, and 75% in 9880 ppm F at pH 5.5.

[0158] A 75% w / v solution was prepared by adding 75 g of CPP-ACP powder to 20 ml of water, each addition being made in small amounts (0.5 g / min) and maintaining the pH at 5.5 by simultaneously adding 10 M HCl. The solution was mixed thoroughly after each addition to ensure dispersion. A concentrated NaF (0.95 M) solution was added along with the 10 M HCl to achieve a final addition of 52 mmol of F. The CPP-ACP powder, NaF, and HCl were added with water over 2–3 h, bringing the final volume to 100 ml. This produced a highly viscous solution of 75% w / v CPP-ACP, 9,880 ppm F, and pH 5.5.

[0159] Example 3 In vitro remineralization by CPP-ACFP and CPP-ACP at high concentrations. The purpose of these experiments was to compare the remineralization by CPP-ACP plus fluoride (F) and CPP-ACP at high concentrations (e.g., 20% w / v, 25% w / v, 30% w / v, 38% w / v, 40% w / v, 50% w / v, and 63% w / v).

[0160] Demineralized subsurface lesions of human dental enamel were created in blocks of third molar enamel. Half of the block was kept as a control, and the other half of the block was treated as follows: Each enamel specimen was pretreated with 1 M NaOH (5 ml) at 45°C for 5 min, then washed with water for 10 s and patted dry; They were remineralized by individual suspension in one of the following remineralization solutions for 4 hours at 45°C: - CPP-ACP+F liquid compositions at 25% w / v, 38% w / v, 50% w / v, and 63% w / v of CPP-ACP; or - Liquid compositions of CPP-ACP only (no fluoride) at 20% w / v, 30% w / v, 40% w / v, and 50% w / v CPP-ACP.

[0161] The fluoride contents for the CPP-ACP+F liquid compositions are shown in Table 2, specifically 3,260 ppm F for 25% w / v, 4,890 ppm F for 38% w / v, 6,520 ppm F for 50% w / v, and 8,151 ppm F for 63% w / v CPP-ACP, respectively.

[0162] Enamel blocks were removed and combined with their controls, embedded, sectioned, and subjected to cross-sectional microradiography and densitometric image analysis to determine percent mineral content increase (% remineralization).

[0163] [Table 3]

[0164] The level of remineralization below the enamel surface in just 4 hours is the highest level of remineralization reported for such a short exposure time and was achieved through a novel formulation of a liquid composition containing a high concentration of CPP-ACP(F) and a novel use of temperature to promote remineralization.

[0165] Example 4 A method for preparing a highly concentrated liquid composition containing CPP-ACP or CPP-ACP and having free fluoride is described below.

[0166] Thirty grams of CPP-ACP powder (commercially available from Recaldent) was added to 19.5 grams of a 20,000 ppm F (NaF) solution, to which 0.5 grams of 11 M HCl solution was added to obtain a final volume of 50 grams. (This final solution is therefore 60% w / w CPP-ACP with 7,800 ppm F at pH 7.8, or 75% w / v CPP-ACP containing 10,000 mg / L F at pH 7.8.) After thorough stirring (approximately 30 minutes), a homogeneous, highly viscous, yet stable solution was prepared at a pH of 7.8. The solution was then degassed by placing it under vacuum for 24 hours to remove trapped air bubbles.

[0167] This viscous, stable, and safe (neutral pH) solution is easy to apply in oral surgery, and the more concentrated it is, the better the effect over a longer period of time. Because the composition is still in liquid form, it can be applied to the tooth surface with a microbrush.

Claims

1. 1. A method of reducing the surface or subsurface visibility of hypomineralized teeth, comprising: contacting the surface or subsurface of the hypomineralized tooth with a liquid composition comprising at least 40% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) and having a pH of 5 or greater but 9 or less; thereby reducing the surface or subsurface visibility of hypomineralized teeth; A method comprising:

2. 1. A method for treating or preventing dentin hypersensitivity in a subject in need thereof, comprising: contacting the exposed dentinal tubules with a liquid composition comprising at least 40% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) at a pH of 5 or greater but 9 or less; thereby treating or preventing dentin hypersensitivity in said subject in need thereof; A method comprising:

3. 3. The method of claim 1 or 2, wherein the pH of the liquid composition is 6 or more but 8 or less, preferably 7 or more but 8 or less.

4. The method of any one of claims 1 to 3, wherein the liquid composition further comprises free fluoride ions.

5. 5. The method of claim 4, wherein the free fluoride ions are present in the liquid composition at a concentration in the range of about 200 ppm to 10,000 ppm.

6. 5. The method of claim 4, wherein the free fluoride ions are present in the liquid composition at a concentration in the range of about 2,600 ppm to about 8,500 ppm.

7. 5. The method of claim 4, wherein said free fluoride ions are present in said liquid composition at a concentration of about 7,800 ppm.

8. The method of any one of claims 1 to 7, wherein the method further comprises applying heat to the surface or subsurface of the tooth or lesion.

9. 9. The method of claim 8, wherein the tooth surface or subsurface is heated to a temperature greater than 37°C.

10. 9. The method of claim 8, wherein the tooth surface or subsurface is heated to a temperature of 40°C or greater.

11. 9. The method of claim 8, wherein the tooth surface or subsurface is heated to a temperature of 45°C or greater.

12. 9. The method of claim 8, wherein the tooth surface or subsurface is heated to a temperature of 50°C or greater.

13. 9. The method of claim 8, wherein the tooth surface or subsurface is heated to a temperature of 55°C or greater.

14. 10. The method of claim 9, wherein the tooth surface or subsurface is heated to a temperature of 60°C or greater.

15. 10. The method of claim 9, wherein the tooth surface or subsurface is heated to a temperature of 65°C or greater.

16. The method of any one of claims 1 to 15, wherein the liquid composition comprises more than 45% w / w stabilized ACP and / or ACFP.

17. The method of any one of claims 1 to 15, wherein the liquid composition comprises more than 50% w / w stabilized ACP and / or ACFP.

18. The method of any one of claims 1 to 15, wherein the liquid composition comprises more than 55% w / w of stabilized ACP and / or ACFP.

19. The method of any one of claims 1 to 15, wherein the liquid composition comprises more than 60% w / w stabilized ACP and / or ACFP.

20. The method of any one of claims 1 to 15, wherein the liquid composition comprises about 65% w / v stabilized ACP and / or ACFP.

21. The method of any one of claims 1 to 15, wherein the liquid composition comprises about 70% w / v stabilized ACP and / or ACFP.

22. The method of any one of claims 1 to 15, wherein the liquid composition comprises about 75% w / v stabilized ACP and / or ACFP.

23. 23. The method of any one of claims 1 to 22, wherein the phosphopeptide is a casein phosphopeptide.

24. 1. A method of reducing the surface or subsurface visibility of hypomineralized teeth, comprising: (i) contacting the surface or subsurface of the hypomineralized tooth with a liquid composition having a pH of 6 or less, the liquid composition comprising at least 40% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); (ii) following (i), increasing the pH of the liquid composition applied to the surface or subsurface of the hypomineralized tooth to about 9 or greater, thereby forming a gel in and / or on the surface or subsurface of the hypomineralized tooth; thereby reducing the surface or subsurface visibility of hypomineralized teeth; A method comprising:

25. 1. A method of forming a gel in and / or on a tooth surface or subsurface lesion, comprising: (i) contacting the tooth surface or subsurface lesion with a liquid composition having a pH of 6 or less, comprising at least 40% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); (ii) subsequent to (i), increasing the pH of the liquid composition applied to the tooth surface or subsurface lesion to about 9 or greater, thereby forming a gel in and / or on the tooth surface or subsurface lesion; thereby forming a gel in and / or on said tooth surface or subsurface lesion; A method comprising:

26. 1. A method for treating or preventing dentin hypersensitivity in a subject in need thereof, comprising: (i) applying at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) to exposed dentinal tubules; a liquid composition having a pH of 6 or less comprising: (ii) following (i), increasing the pH of the liquid composition applied to the surface or subsurface of the hypomineralized tooth to about 9 or greater, thereby forming a gel in and / or on the exposed dentinal tubules; thereby treating or preventing dentin hypersensitivity in said subject in need thereof; A method comprising:

27. 1. A method for forming a protective layer on a tooth surface, comprising: (i) contacting the tooth surface with a liquid composition having a pH of 6 or less, comprising at least 40% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); (ii) subsequent to (i), increasing the pH of the liquid composition applied to the tooth surface to about 9 or greater; thereby forming a protective layer on the tooth surface; A method comprising:

28. 28. The method of any one of claims 24 to 27, wherein the pH of the liquid composition applied to the tooth surface or subsurface lesion is increased to about 10 or greater.

29. 28. The method of any one of claims 24 to 27, wherein the step of increasing the pH of the liquid composition applied to the tooth surface or subsurface lesion is carried out by contacting the liquid composition applied to the tooth surface or subsurface with a further composition of alkaline pH.

30. 30. The method of any one of claims 24 to 29, wherein the tooth surface is tooth enamel.

31. A method according to any one of claims 24 to 30, wherein the tooth surface is a lesion in the enamel caused by caries, dental erosion or fluorosis.

32. 32. The method of claim 31 , wherein the lesion is a vitiligo lesion.

33. 33. The method of any one of claims 24 to 32, wherein the step of increasing the pH of the liquid composition applied to the tooth surface or subsurface lesion is carried out by contacting the liquid composition applied to the tooth surface or subsurface with a further composition of alkaline pH.

34. 1. A method of reducing the surface or subsurface visibility of hypomineralized teeth, comprising: (i) mixing (a) a liquid composition having a pH of 6 or less, comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); and (b) a further composition having an alkaline pH, thereby forming a mixed composition having a pH of about 9 or greater; (ii) contacting the mixed composition with the surface or subsurface of the hypomineralized tooth, thereby forming a gel in and / or on the surface or subsurface of the hypomineralized tooth; thereby reducing the visibility of the hypomineralized tooth surface or subsurface; A method comprising:

35. 1. A method for treating or preventing dentin hypersensitivity in a subject in need thereof, comprising: (i) (a) a pH 6 solution containing at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); (b) mixing a liquid composition having a pH of about 9 or greater with an additional composition having an alkaline pH, thereby forming a mixed composition having a pH of about 9 or greater; (ii) contacting the mixed composition with the exposed dentinal tubules, thereby forming a gel in and / or on the exposed dentinal tubules; thereby treating or preventing dentin hypersensitivity in said subject in need thereof; A method comprising:

36. 1. A method for forming a protective layer on a tooth surface, comprising: (i) mixing (a) a liquid composition having a pH of 6 or less, comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); and (b) a further composition having an alkaline pH, thereby forming a mixed composition having a pH of about 9 or greater; (ii) contacting the mixed composition with the tooth surface; thereby forming a protective layer on the tooth surface; A method comprising:

37. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 8.

38. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 9.

39. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 10.

40. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 11.

41. 37. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 12.

42. 37. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 13.

43. The method of any one of claims 34 to 36, wherein the alkaline pH of the further composition is a pH of about 14.

44. The method of any one of claims 24 to 43, wherein the liquid composition further comprises free fluoride ions.

45. 45. The method of claim 44, wherein the free fluoride ions are present in the liquid composition at a concentration in the range of about 200 ppm to about 10,000 ppm.

46. 45. The method of claim 44, wherein the free fluoride ions are present in the liquid composition at a concentration in the range of about 2,600 ppm to about 8,500 ppm.

47. 45. The method of claim 44, wherein the free fluoride ions are present in the liquid composition at a concentration of about 8,200 ppm.

48. 34. The method of any one of claims 24 to 33, wherein step (i) further comprises applying heat to the surface or subsurface of the tooth or lesion.

49. 34. A method according to any one of claims 24 to 33, wherein step (i) is followed by a step of heating the surface or subsurface of the tooth or lesion.

50. 50. The method of any one of claims 24 to 49, wherein step (ii) further comprises applying heat to the surface or subsurface of the tooth or lesion.

51. 50. A method according to any one of claims 24 to 49, wherein step (ii) is followed by a step of heating the surface or subsurface of the tooth or lesion.

52. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature above 37°C.

53. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of 40°C or above.

54. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of 45°C or greater.

55. The surface or subsurface of the tooth or lesion is heated to a temperature of 50°C or greater.

52. The method of any one of claims 48 to 51.

56. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of 55°C or greater.

57. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of 60°C or greater.

58. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of 65°C or greater.

59. 52. A method according to any one of claims 48 to 51, wherein the surface or subsurface of the tooth or lesion is heated to a temperature of less than 65°C.

60. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises more than 45% w / v of stabilized ACP and / or ACFP.

61. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises more than 50% w / v of stabilized ACP and / or ACFP.

62. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises more than 55% w / v of stabilized ACP and / or ACFP.

63. The liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises more than 60% w / v of stabilized ACP and / or ACFP. Item 60. The method according to any one of Items 24 to 59.

64. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises about 65% w / v of stabilized ACP and / or ACFP.

65. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises about 70% w / v of stabilized ACP and / or ACFP.

66. 60. The method of any one of claims 24 to 59, wherein the liquid composition comprising more than 40% w / v of phosphopeptide (PP) stabilized ACP and / or ACFP comprises about 75% w / v of stabilized ACP and / or ACFP.

67. 34. The method of any one of claims 24 to 33, wherein the liquid composition in (i) is contacted with the surface or subsurface of the hypomineralized tooth or lesion for up to 20 minutes before the step of increasing the pH of the liquid composition applied to the surface or subsurface of the hypomineralized tooth to about 9 or greater.

68. 68. The method of claim 67, wherein the liquid composition in (i) is contacted with the hypomineralized tooth surface or subsurface or lesion for at least about several seconds to about 5 minutes.

69. 69. The method of claim 68, wherein the liquid composition in (i) is contacted with the hypomineralized tooth surface or subsurface or lesion for at least about 5 minutes to about 20 minutes.

70. 70. The method of any one of claims 24 to 69, wherein the method further comprises the step of acid etching the hypomineralized surface, subsurface, or lesion prior to contacting with the liquid composition in (i).

71. 71. The method of any one of claims 24 to 70, wherein the phosphopeptide is a casein phosphopeptide.

72. 72. The method of any one of claims 1 to 71, wherein the liquid, further composition and / or mixed composition is applied to the surface, subsurface or lesion of the tooth by a dental health care professional.

73. 73. The method of claim 72, wherein the liquid, further composition, and / or mixed composition is applied to the tooth surface, subsurface, or lesion using a microbrush.

74. The method further comprises identifying a subject having a white spot lesion, a fluorosis lesion, a caries lesion, or a lesion caused by dental erosion.

74. The method of any one of claims 1 to 73.

75. 1. A cosmetic method for reducing the surface or subsurface visibility of hypomineralized teeth, comprising: (i) contacting the surface or subsurface of the hypomineralized tooth with a liquid composition comprising at least 40% w / v phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) at a pH of 6 or less; (ii) Following (i), increasing the pH of the liquid composition applied to the surface or subsurface of the hypomineralized tooth to about 9 or greater, thereby reducing the surface or subsurface pH of the hypomineralized tooth. forming a gel therein and / or thereon; thereby reducing the surface or subsurface visibility of hypomineralized teeth; Beauty methods, including:

76. 1. A cosmetic method for reducing the surface or subsurface visibility of hypomineralized teeth, comprising: contacting the surface or subsurface of the hypomineralized tooth with a liquid composition having a pH of 6 or greater but not greater than 8, the liquid composition comprising at least 40% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); thereby reducing the visibility of the hypomineralized tooth surface or subsurface; Beauty methods, including:

77. 1. A kit for reducing the surface or subsurface visibility of hypomineralized teeth, comprising or consisting of: (a) a liquid composition having a pH of 6 or less, comprising at least 40% by weight of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP); and (b) a further composition having an alkaline pH.

78. 78. The kit of claim 77, further comprising written instructions for use in the method of any one of claims 1 to 76.

79. 1. A kit for reducing the surface or subsurface visibility of hypomineralized teeth, comprising: (a) a first composition comprising a powder of phosphopeptide-stabilized ACP and / or ACFP; (b) a second composition having a pH of 6 or less, comprising a solution of fluoride; (c) a third composition at an alkaline pH; and A kit comprising or consisting of:

80. 80. The kit of any one of claims 77-79, wherein the alkaline pH of the third composition is a pH of about 9, 10, 11, 12, 13, or 14.

81. 79. The kit of claim 77 or 78, wherein the first composition contains an amount of phosphopeptide-stabilized ACP and / or ACFP that, when mixed with the second composition, forms a liquid composition containing at least 40% w / v of phosphopeptide-stabilized ACP and / or ACFP.

82. 1. A kit for reducing the surface or subsurface visibility of hypomineralized teeth, comprising: (a) 5 g of CPP-ACP and / or CPP-ACFP; (b) 5 ml of 0.73 M NaF in 1.146 M HCl; (c) 1.5 M NaOH; A kit comprising or consisting of:

83. 83. The kit of claim 82, further comprising two microbrushes.

84. 1. A method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or ACFP, comprising: mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP; maintaining the pH below 7; A method or process comprising or consisting of:

85. 1. A method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or ACFP, comprising: mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP; reducing the pH to less than 7, preferably to a pH of 6 or less, preferably 5.5 or less; A method or process comprising or consisting of:

86. 86. The method or process of claim 84 or 85, wherein the pH is maintained at or below 6, preferably the pH is maintained at or below 5.

5.

87. To prepare a powder comprising or consisting of PP-stabilized ACP and / or ACFP: mixing one or more solutions comprising phosphopeptides, calcium ions, phosphate ions, hydroxide ions, and optionally fluoride ions while maintaining a pH of about 7.0 or greater, preferably about 9, to form a solution comprising stabilized ACP and / or ACFP; drying the solution containing PP-stabilized ACP and / or ACFP; thereby forming a powder comprising or consisting of PP-stabilized ACP and / or ACFP; 87. The method or process of any one of claims 84 to 86, further comprising:

88. 88. The method or process of claim 87, wherein the drying is spray drying or freeze drying.

89. 89. The method or process of claim 87 or 88, further comprising filtering the solution comprising PP-stabilized ACP and / or ACFP to form a retentate prior to drying, and subsequently drying the retentate to form a powder comprising or consisting of PP-stabilized ACP and / or ACFP.

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