Hiddenness of hypomineralized lesions

Abstract

The present invention relates to compositions and uses for reducing the visibility of hypomineralized tooth surfaces or subsurfaces, particularly tooth enamel. In one aspect, the present invention relates to a method for reducing the visibility of hypomineralized tooth surfaces or subsurfaces, the method comprising (i) contacting a hypomineralized tooth surface or subsurface with a liquid composition comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and (ii) after (i), raising the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface to equal to or greater than about 9, thereby forming a gel in and / or on the hypomineralized tooth surface or subsurface, thereby reducing the visibility of the hypomineralized tooth surface or subsurface.

Description

[0001] Cross-reference to prior applications

[0002] This application claims priority to Australian provisional applications Nos. 2019900833 and 2019903860, the contents of each of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention relates to compositions and uses for reducing the visibility of hypomineralised tooth surfaces or sub-surfaces, particularly tooth enamel. Background Art

[0004] White spots are porous lesions in the enamel / dentin that can result from caries (white spot lesions), hypomineralisation (e.g. fluorosis and other developmental defects) or other demineralisation processes (e.g. erosion lesions). Typically, they represent an early stage in caries development, where the affected surface appears intact after mild probing. Other common causes of hypomineralisation and associated white spot lesions include trauma, xerostomia and arrested decay that is only partially remineralised around fixed orthodontic appliances, which may provide harbour for bacteria or interfere with normal remineralisation.

[0005] This porous tissue in enamel and dentin has a different refractive index than the translucent enamel and dentin, so porous lesions appear as opaque white lesions against a translucent background. These lesions are not only a cosmetic problem, but also increase the risk of caries / erosion development and hypersensitivity reactions.

[0006] Current treatment of these lesions prior to cavitation involves the use of fluoride for remineralization (with or without calcium phosphate), but remineralization is a slow process that takes months to change the appearance of the lesion, which often never completely disappears. Other treatments involve invasive or at best minimally invasive restorations, including infiltration with methacrylate-based resins. These restorative procedures are expensive, time-consuming, and do not allow for the natural restoration of the white spots (remineralization with calcium and phosphates), which can lead to subsequent problems with discoloration of the restorative material or caries beneath the restoration due to poor sealing.

[0007] The best treatment for repairing porous leukoplakia and reducing sensitivity is the use of CPP-ACP (commercially available as Recaldent TMLesions can be treated with fluoride (with or without fluoride) to achieve remineralization using hydroxyapatite or fluorohydroxyapatite. However, current treatments using this technology still require several weeks of treatment at best to improve the appearance of the lesion and reduce sensitivity. This can lead to poor patient outcomes due to low compliance and insufficient remineralization.

[0008] There is a need to provide new or improved methods for reducing the visibility of and / or forming a barrier to hypomineralized tooth surfaces or subsurfaces.

[0009] The reference to any prior art in the specification is not an acknowledgement or implication that the prior art forms part of the common general knowledge in Australia or any other jurisdiction, or that the prior art could reasonably be ascertained, understood and regarded as relevant by a person skilled in the art. Summary of the invention

[0010] In one aspect, the present invention provides a method of reducing the visibility of a hypomineralized tooth surface or subsurface, the method comprising:

[0011] contacting a hypomineralized tooth surface or subsurface with a liquid composition comprising at least 40% w / w phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9,

[0012] This reduces the visibility of hypomineralized tooth surfaces or subsurfaces.

[0013] Preferably, the pH of the liquid composition is greater than or equal to pH 6 but less than or equal to pH 8, such as greater than or equal to pH 7 but less than or equal to pH 8.

[0014] In another aspect, the present invention provides a method for forming a protective layer on a tooth surface, the method comprising:

[0015] contacting the tooth surface with a liquid composition comprising at least 40% w / w of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9,

[0016] This forms a protective layer on the tooth surface.

[0017] Preferably, the pH of the liquid composition is greater than or equal to pH 6 but less than or equal to pH 8, such as greater than or equal to pH 7 but less than or equal to pH 8.

[0018] In another aspect, the present invention provides a method of treating or preventing dentin hypersensitivity in a subject in need thereof, the method comprising:

[0019] contacting the exposed dentinal tubules, contacting the tooth surface with a liquid composition, the liquid composition comprising at least 40% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9,

[0020] Thereby treating or preventing dentin hypersensitivity in a subject in need thereof.

[0021] Preferably, the pH of the liquid composition is greater than or equal to pH 6 but less than or equal to pH 8, such as greater than or equal to pH 7 but less than or equal to pH 8.

[0022] Preferably, a heat source is applied to the liquid composition once the liquid composition contacts the tooth surface or subsurface.

[0023] In one aspect, the present invention provides a method of reducing the visibility of a hypomineralized tooth surface or subsurface, the method comprising:

[0024] (i) contacting a hypomineralized tooth surface or subsurface with a liquid composition comprising at least 40% w / v phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and

[0025] (ii) after (i), raising the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface to a pH of equal to or greater than about 9, thereby forming a gel in and / or on the hypomineralized tooth surface or subsurface,

[0026] This reduces the visibility of hypomineralized tooth surfaces or subsurfaces.

[0027] In another aspect, the present invention provides a method of forming a gel within and / or on a tooth surface or subsurface lesion, the method comprising:

[0028] (i) contacting a tooth surface or subsurface lesion with a liquid composition comprising at least 40% w / v phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and

[0029] (ii) after (i), raising the pH of the liquid composition applied to the tooth surface or subsurface lesion to a pH of equal to or greater than about 9,

[0030] A gel is thereby formed within and / or on the tooth surface or subsurface lesions.

[0031] In another aspect, the present invention provides a method of reducing the visibility of a hypomineralized tooth surface or subsurface, the method comprising:

[0032] (i) mixing (a) a liquid composition and (b) another composition having an alkaline pH, thereby forming a mixed composition having a pH equal to or greater than about 9, wherein the liquid composition comprises at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), and the pH of the liquid composition is less than or equal to pH 6;

[0033] (ii) contacting a hypomineralized tooth surface or subsurface with the mixed composition, thereby forming a gel in and / or on the hypomineralized tooth surface or subsurface,

[0034] This reduces the visibility of hypomineralized tooth surfaces or subsurfaces.

[0035] In another aspect, the present invention provides a method of treating or preventing dentin hypersensitivity in a subject in need thereof, the method comprising:

[0036] (i) contacting the exposed dentinal tubules with a liquid composition comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and

[0037] (ii) after (i), raising the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface to a pH of equal to or greater than about 9, thereby forming a gel within and / or on the exposed dentinal tubules,

[0038] Thereby treating or preventing dentin hypersensitivity in a subject in need thereof.

[0039] In another aspect, the present invention provides a method of treating or preventing dentin hypersensitivity in a subject in need thereof, the method comprising:

[0040] (i) mixing (a) a liquid composition and (b) another composition having an alkaline pH, thereby forming a mixed composition having a pH equal to or greater than about 9, wherein the liquid composition comprises at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), and the pH of the liquid composition is less than or equal to pH 6;

[0041] (ii) contacting the exposed dentinal tubules with the mixed composition, thereby forming a gel in and / or on the exposed dentinal tubules,

[0042] Thereby treating or preventing dentin hypersensitivity in a subject in need thereof.

[0043] In another aspect, the present invention provides a method for forming a protective layer on a tooth surface, the method comprising:

[0044] (i) contacting the tooth surface with a liquid composition comprising at least 40% w / v phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and

[0045] (ii) after (i), raising the pH of the liquid composition applied to the tooth surface to a pH equal to or greater than about 9,

[0046] This forms a protective layer on the tooth surface.

[0047] In another aspect, the present invention provides a method for forming a protective layer on a tooth surface, the method comprising:

[0048] (i) mixing (a) a liquid composition and (b) another composition having an alkaline pH, thereby forming a mixed composition having a pH equal to or greater than about 9, wherein the liquid composition comprises at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), and the pH of the liquid composition is less than or equal to pH 6;

[0049] (ii) contacting the tooth surface with the mixed composition,

[0050] This forms a protective layer on the tooth surface.

[0051] In any aspect of the invention, the pH of the liquid composition applied to the tooth surface or subsurface lesion is raised to a pH equal to or greater than, about 9, preferably 10.

[0052] In any aspect of the invention, the tooth surface is preferably tooth enamel. In one embodiment, the tooth surface is a lesion in tooth enamel, such as a lesion caused by caries, dental erosion or fluorosis. Typically, the lesion is a white spot lesion.

[0053] In any aspect of the invention, the tooth surface is dentin, such as exposed dentin. In one embodiment, the exposed dentin is a tooth root, such as caused by degeneration.

[0054] In any aspect of the invention, the tooth surface is contacted with the mixed composition less than 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 minute after the mixed composition is formed.

[0055] In any aspect of the present invention, in the case where it is necessary to increase the pH of a liquid composition applied to a tooth surface or subsurface, this can be accomplished by contacting the liquid composition applied to the tooth surface or subsurface with another composition having an alkaline pH. The other composition may also be in liquid form and may also be referred to herein as another liquid composition. The alkaline pH of the other composition may be about 9, 10, 11, 12, 13 or 14. Preferably, the other composition has an alkaline pH of about 9 or more.

[0056] In any aspect described herein, the liquid composition may also contain fluoride ions, preferably free fluoride ions. Fluoride ions may be present in the liquid composition at a concentration in the range of about 200 ppm to 50,000 ppm. In a preferred embodiment, the concentration of fluoride ions is in the range of about 2,600 ppm to about 10,000 ppm. In a further preferred embodiment, the fluoride ion concentration in the liquid composition is about 8,200 ppm or about 6,500 ppm. Fluoride ions may be present in the liquid composition at a concentration equal to or greater than any ppm described herein (particularly in the embodiments). In another embodiment, the concentration of fluoride ions is about 5,200 ppm for 40% w / v CPP-ACP; about 5,850 ppm for 45% w / v CPP-ACP; about 6,500 ppm for 50% w / v CPP-ACP; about 8,200 ppm for 63% w / v CPP-ACP; or about 9,900 ppm for 75% CPP-ACP. In another embodiment, the fluoride ion concentration is about 5,200 ppm for 40% w / w CPP-ACP or about 7,800 ppm for 60% w / w CPP-ACP. The fluoride ions can come 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, the following: sodium fluoride, sodium monofluorophosphate, stannous fluoride, sodium fluorosilicate, silver fluoride, silver diamine fluoride, and amine fluorides. These may be provided in the form of a solution (typically an aqueous solution) or a suspension.

[0057] In any aspect of the invention, steps (i) and (ii), or any step of applying a liquid composition to a tooth surface or subsurface, may include or be followed by heating the tooth surface or subsurface or lesion. In one embodiment, the method may include heating the hypomineralized tooth surface or subsurface or lesion while or after contacting the liquid composition with the hypomineralized tooth surface or subsurface or lesion, the liquid composition comprising at least 40% w / v of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP, the pH of the liquid composition being less than or equal to pH 6. In another embodiment, the method may include heating the hypomineralized tooth surface or subsurface or lesion while or after raising the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface to equal to or greater than about 9. In another embodiment, the method may comprise heating the hypomineralized dental surface or subsurface or lesion simultaneously with or after contacting the hypomineralized dental surface or subsurface or lesion with a liquid composition, wherein the liquid composition comprises at least 60% w / w of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP, and the pH of the liquid composition is greater than or equal to pH 5 but less than or equal to pH 9.

[0058] In any aspect of the invention, the method comprises heating a tooth surface or subsurface or lesion to a temperature greater than or equal to 40°C, greater than or equal to 45°C, greater than or equal to 50°C, greater than or equal to 55°C, greater than or equal to 60°C or greater than or equal to 65°C.

[0059] In any aspect of the invention, the method comprises heating a tooth surface or subsurface or lesion to a temperature greater than 37°C but equal to or less than 65°C, greater than 40°C but equal to or less than 65°C, greater than 45°C but equal to or less than 65°C, greater than 50°C but equal to or less than 65°C, greater than 55°C but equal to or less than 65°C, greater than 60°C but equal to or less than 65°C.

[0060] In any aspect of the invention, the liquid composition comprising greater than 40% w / v of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP comprises greater than 45% w / v, greater than 50% w / v of stable ACP and / or stable ACFP, greater than 55% w / v of stable ACP and / or stable ACFP, greater than 60% w / v of stable ACP and / or stable ACFP, greater than about 65% w / v of stable ACP and / or stable ACFP, greater than about 70% w / v of stable ACP and / or stable ACFP, or greater than about 75% w / v of stable ACP and / or stable ACFP. In one embodiment, the liquid composition comprises 63% w / v of stable ACP and / or stable ACFP.

[0061] In any aspect of the invention, the liquid composition comprising greater than 40% w / w of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP comprises greater than 45% w / w, greater than 50% w / w of stabilized ACP and / or stabilized ACFP, greater than 55% w / w of stabilized ACP and / or stabilized ACFP, greater than 60% w / w of stabilized ACP and / or stabilized ACFP, greater than about 65% w / w of stabilized ACP and / or stabilized ACFP, greater than about 70% w / w of stabilized ACP and / or stabilized ACFP, or greater than about 75% w / w of stabilized ACP and / or stabilized ACFP.

[0062] In any aspect of the invention, the liquid composition comprising greater than 40% w / v of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP comprises greater than 40% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 45% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 50% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 55% w / v of stable ACP and / or stable ACFP, The invention also provides a stable ACP and / or stable ACFP having a stable ACFP content of greater than 60% w / v but less than 80% w / v of stable ACP and / or stable ACFP, a stable ACP and / or stable ACFP having a stable ACFP content of greater than 65% w / v but less than 80% w / v of stable ACP and / or stable ACFP, a stable ACP and / or stable ACFP having a stable ACFP content of greater than 70% w / v but less than 80% w / v of stable ACP and / or stable ACFP, a stable ACP and / or stable ACFP having a stable ACFP content of greater than 75% w / v but less than 80% w / v of stable ACP and / or stable ACFP.

[0063] In any aspect of the invention, the liquid composition comprising greater than 40% w / w of phosphopeptide (PP)-stabilized ACP and / or phosphopeptide (PP)-stabilized ACFP comprises greater than 40% w / w of stabilized ACP and / or stabilized ACFP but less than 80% w / w of stabilized ACP and / or stabilized ACFP, greater than 45% w / w of stabilized ACP and / or stabilized ACFP but less than 80% w / w of stabilized ACP and / or stabilized ACFP, greater than 50% w / w of stabilized ACP and / or stabilized ACFP but less than 80% w / w of stabilized ACP and / or stabilized ACFP, greater than 55% w / w of stabilized ACP and / or stabilized ACFP, The invention also provides a stable ACP and / or stable ACFP having a stable ACP and / or stable ACFP of at least 80% w / w but less than 80% w / w, a stable ACP and / or stable ACFP having a stable ACP and / or stable ACFP of at least 60% w / w but less than 80% w / w, a stable ACP and / or stable ACFP of at least 65% w / w but less than 80% w / w, a stable ACP and / or stable ACFP of at least 70% w / w but less than 80% w / w, a stable ACP and / or stable ACFP of at least 75% w / w but less than 80% w / w.

[0064] In any aspect of the invention, the liquid composition is contacted with the hypomineralized dental surface or subsurface or lesion in (i) for a period of time to allow the liquid composition to penetrate the dental surface, subsurface or lesion. In one embodiment, the hypomineralized dental surface or subsurface or lesion is contacted with the liquid composition in (i) for up to 20 minutes before the pH of the liquid composition applied to the hypomineralized dental surface or subsurface is raised to equal to or greater than about 9. In one embodiment, the hypomineralized dental surface or subsurface or lesion is contacted with the liquid composition in (i) for at least about a few seconds to at least about 5 minutes, preferably at least about 5 minutes to about 20 minutes.

[0065] In any aspect of the invention, prior to contact with the liquid composition (e.g., in (i)), the method further comprises etching, preferably acid etching, the hypomineralized surface, subsurface or lesion. Acid etching can be performed by any method known in the art. Typically, acid etching can be performed by contacting the hypomineralized surface, subsurface or lesion with a composition comprising about 30% phosphoric acid or about 15% HCl. Preferably, a resin barrier is applied prior to acid etching to protect the soft tissue, such as a rubber dam or a liquid rubber dam.

[0066] In any aspect, the formation of the protective layer may be facilitated by the further steps of heating as described herein and / or etching as described herein.

[0067] The stabilized ACP and / or stabilized ACFP used herein is stabilized by a phosphopeptide. 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.

[0068] In a preferred embodiment, the phosphopeptide-stabilized amorphous calcium phosphate (ACP) or phosphopeptide-stabilized amorphous calcium fluoride phosphate (ACFP) complex has tightly bound and loosely bound calcium, wherein the bound calcium in the complex is less than the tightly bound calcium in the ACP or ACFP complex formed at a pH of 7.0. Typically, the ACP or ACFP is primarily in the alkaline form.

[0069] In a preferred embodiment, the calcium ion content of the stabilized ACP or stabilized ACFP complex 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.

[0070] In any aspect or embodiment as described herein, the stabilized ACP and / or stabilized ACFP may be in a formulation with additional calcium phosphate. Typically, the formulation includes a PP-stabilized ACP and / or PP-stabilized ACFP complex and at least an equal weight of calcium phosphate.

[0071] In a preferred embodiment, the PP-stabilized ACP and / or PP-stabilized ACFP is in the form of a casein phosphopeptide-stabilized ACP and / or ACFP complex.

[0072] Preferably, the phase of ACP is mainly an alkaline phase, wherein ACP mainly comprises Ca 2+ PO4 3- and OH - The alkaline phase of ACP may have the general formula [Ca3(PO4)2] x [Ca2(PO4)(OH)], wherein x≥1. Preferably x=1-5. More preferably, x=1. Preferably, the two components in the formula are present in equal proportions. Thus, in one embodiment, the alkaline phase of the ACP has the chemical formula Ca3(PO4)2Ca2(PO4)(OH).

[0073] Preferably, the phase of ACFP is mainly an alkaline phase, wherein ACFP mainly comprises Ca 2+ PO4 3- and F - The basic phase of ACFP can have the general formula [Ca3(PO4)2] x [Ca2(PO4)F] y, wherein x≥1 when y=1 or y≥1 when x=1. Preferably, y=1 and x=1-3. More preferably, y=1 and x=1. Preferably, the two components of the formula are present in equal proportions. Thus, in one embodiment, the basic phase of ACFP has the chemical formula Ca3(PO4)2Ca2(PO4)F.

[0074] In one embodiment, the ACP complex consists essentially of the phosphopeptide, calcium, phosphate and hydroxide ions, and water.

[0075] In one embodiment, the ACFP complex consists essentially of the phosphopeptide, calcium, phosphate, fluoride and hydroxide ions, and water.

[0076] In another aspect of the present invention, there is provided a method for remineralizing a dental lesion, the method comprising:

[0077] (i) contacting a dental lesion with a liquid composition comprising at least 40% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6; and

[0078] (ii) after (i), raising the pH of the liquid composition applied to the dental lesion to a pH equal to or greater than about 9, thereby forming a gel within and / or on the dental lesion,

[0079] Thereby remineralizing the dental lesions.

[0080] In another aspect of the present invention, there is provided a method of remineralizing a dental lesion, the method comprising:

[0081] contacting a hypomineralized tooth surface or subsurface with a liquid composition comprising at least 40% w / w phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9,

[0082] Thereby remineralizing the dental lesions.

[0083] Preferably, the pH of the liquid composition is greater than or equal to pH 6 but less than or equal to pH 8, such as greater than or equal to pH 7 but less than or equal to pH 8.

[0084] Preferably, the dental lesion is in the enamel or dentin. Typically, the dental lesion is selected from the group consisting of one or more of: a white spot lesion; a fluorosis lesion; a carious lesion; or a lesion caused by dental erosion.

[0085] In any aspect or embodiment of the invention described herein, the liquid compositions and other compositions or mixed compositions as described herein are applied to the mouth, teeth or lesions of a subject in need of treatment by a dental health professional.

[0086] In one embodiment, the tooth surface is in need of such treatment. Thus, in addition to the steps of any of the methods described herein, the present invention also includes the step of identifying a subject suffering from fluorosis, dental caries, dentin hypersensitivity, or dental calculus, white spot lesions, fluorosis, carious lesions, or lesions caused by dental erosion. Specifically, the tooth surface in need of forming a protective layer may be exposed dentin, which typically results in dentin hypersensitivity in the subject. Exposed dentin may have exposed dentinal tubules.

[0087] Typically, the tooth surface may be one that has been determined to benefit from a surface layer due to, for example, an increased likelihood of demineralization.

[0088] The method of the present invention for forming a protective layer on a tooth surface is particularly useful for sealing exposed dentin, particularly exposed dentin having exposed dentinal tubules.

[0089] The tooth surface may also be a cavity, whereby the methods of the invention described herein are applied to the cavity, thereby allowing a gel or protective layer to form. A dental restorative such as a composite material or glass ionomer cement may then be added to the gel or protective layer.

[0090] In another aspect, the present invention provides a method of repairing a tooth cavity, the method comprising:

[0091] (i) forming a gel or protective layer in the cavity by any of the methods described herein; and

[0092] (ii) applying dental restoratives,

[0093] Thus repairing tooth cavities.

[0094] The present invention provides a liquid composition for mineralizing a tooth surface or subsurface and an additional composition at an alkaline pH, wherein the liquid composition comprises at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, and the additional composition being applied to the tooth surface or subsurface after the first composition, thereby forming a gel.

[0095] The present invention provides a liquid composition comprising at least 40% w / w of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9, the liquid composition being used for:

[0096] Forming layers on the tooth surface,

[0097] Gel formation in and / or on surface or subsurface lesions of teeth,

[0098] Treat or prevent dentin hypersensitivity,

[0099] Reduce the visibility of hypomineralized tooth surfaces or subsurfaces, or

[0100] Remineralize tooth surfaces or subsurfaces.

[0101] Preferably, the pH of the liquid composition is greater than or equal to pH 6 but less than or equal to pH 8, such as greater than or equal to pH 7 but less than or equal to pH 8.

[0102] In another aspect, there is provided a method for treating or preventing one or more of dental caries, tooth decay, dental erosion, fluorosis and white spot lesions, comprising the steps of:

[0103] (i) contacting a tooth surface or subsurface with a liquid composition comprising at least 40 wt% of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6;

[0104] (ii) after (i), raising the pH of the liquid composition applied to the dental lesion to a pH equal to or greater than about 9, thereby causing the first composition to form a gel.

[0105] Topical administration of the composition is preferred. The method preferably comprises administering the complex in a formulation as described above.

[0106] In another aspect, there is provided a use of a phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or a phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP) for the manufacture of a product comprising a liquid composition for reducing the visibility of a hypomineralized tooth surface or subsurface, the liquid composition comprising at least 40% w / v of a phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or a phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6,

[0107] wherein a liquid composition is applied to a hypomineralized tooth surface or subsurface, and then the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface is raised to equal to or greater than about 9, thereby forming a gel in and / or on the hypomineralized tooth surface or subsurface.

[0108] In another aspect, there is provided a use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP) for the manufacture of a product comprising a liquid composition for forming a gel in a tooth surface or subsurface lesion, the liquid composition comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6,

[0109] wherein a liquid composition is applied to a tooth surface or subsurface lesion, and then the pH of the liquid composition applied to the tooth surface or subsurface lesion is raised to equal to or greater than about 9, thereby forming a gel in / on the tooth surface or subsurface lesion.

[0110] In another aspect, there is provided the use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising or consisting of a liquid composition for:

[0111] Forming layers on the tooth surface,

[0112] Gel formation in and / or on surface or subsurface lesions of teeth,

[0113] Treat or prevent dentin hypersensitivity,

[0114] Reduce the visibility of hypomineralized tooth surfaces or subsurfaces, or

[0115] Remineralize tooth surfaces or subsurfaces.

[0116] The liquid composition comprises at least 40% w / w phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), and the pH of the liquid composition is greater than or equal to pH 5 but less than or equal to pH 9. In one embodiment, the product is a cosmetic product.

[0117] In another aspect, there is provided the use of a base in the preparation of a composition for reducing the visibility of a hypomineralized tooth surface or subsurface in a subject to which a liquid composition has been applied, the liquid composition comprising at least 40% w / v phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6.

[0118] In another aspect, there is provided the use of a base in the preparation of a composition for forming a gel in and / or on a surface or subsurface lesion of a tooth of a subject to which a liquid composition has been applied, the liquid composition comprising at least 40% w / v phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6.

[0119] In another aspect, there is provided a use of a phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or a phosphopeptide (PP) stabilized amorphous calcium phosphate (ACFP) and a base in the manufacture of a product comprising a liquid composition and a further composition, the liquid composition comprising at least 40% w / v of a phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or a phosphopeptide (PP) stabilized amorphous calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, the further composition further comprising the base, the liquid composition and the further composition being used to reduce the visibility of a hypomineralized tooth surface or subsurface, wherein after the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface is raised to equal to or greater than about 9, the further composition is applied to the hypomineralized tooth surface or subsurface, thereby forming a gel in and / or on the hypomineralized tooth surface, thereby reducing the visibility of the hypomineralized tooth surface or subsurface. In one embodiment, the product is a cosmetic product.

[0120] In another aspect, there is provided a use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP) and a base in the manufacture of a product comprising a liquid composition and another composition, the liquid composition comprising at least 40% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, the other composition comprising the base, and the liquid composition and the other composition being used to form a gel in and / or on a tooth surface or subsurface lesion, wherein after the pH of the liquid composition applied to the tooth surface or subsurface lesion is raised to equal to or greater than about 9, the other composition is applied to the tooth surface or subsurface lesion, thereby forming a gel in and / or on the tooth surface or subsurface lesion.

[0121] In a preferred embodiment, the phosphopeptide-stabilized amorphous calcium phosphate (ACP) or phosphopeptide-stabilized amorphous calcium fluoride phosphate (ACFP) complex in the composition has tightly bound and loosely bound calcium, wherein the bound calcium in the complex is 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 primarily in alkaline form.

[0122] In another preferred embodiment, the calcium ion content of the stable ACP or stable ACFP complex in the composition 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.

[0123] In any embodiment, the ACP and / or ACFP in the composition may be in the form of a casein phosphopeptide-stabilized ACP and / or ACFP complex.

[0124] On the other hand, the present invention provides a liquid composition comprising at least 40% w / v of stable ACP and / or stable ACFP. Preferably, the liquid composition comprises a stable ACP and / or stable ACFP greater than or equal to 45% w / v, a stable ACP and / or stable ACFP greater than or equal to 50% w / v, a stable ACP and / or stable ACFP greater than or equal to 55% w / v, a stable ACP and / or stable ACFP greater than or equal to 60% w / v, a stable ACP and / or stable ACFP greater than or equal to 65% w / v, a stable ACP and / or stable ACFP greater than or equal to 70% w / v, a stable ACP and / or stable ACFP greater than or equal to 75% w / v. In one embodiment, the liquid composition comprises 63% w / v of stable ACP and / or stable ACFP. Preferably, the liquid composition also comprises fluoride ions as described herein.

[0125] In this aspect of the invention, the liquid composition may comprise greater than 40% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 45% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 50% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, greater than 55% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, or stable ACFP, more than 60% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, more than 65% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, more than 70% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP, more than 75% w / v of stable ACP and / or stable ACFP but less than 80% w / v of stable ACP and / or stable ACFP. Preferably, the liquid composition further comprises fluoride ions as described herein.

[0126] In any aspect, the liquid composition is degassed. Degassing can be performed by any method that creates a negative pressure above the liquid composition. Exemplary methods include vacuum pumps or systems, such as a venturi vacuum water system.

[0127] Any composition described herein can be used in any of the methods described herein. The composition is a physiologically acceptable composition as described herein.

[0128] In any method or use of the invention described herein, the method or use is for the cosmetic purpose of disguising or concealing the visual appearance of a hypomineralised surface or sub-surface.

[0129] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or ACFP, the method or process comprising or consisting of the following steps:

[0130] mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP, and

[0131] The pH is maintained at a value below 7. Preferably, the pH is maintained at a value at or below 6, preferably at a pH of 5.5 or below.

[0132] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or PP-stabilized ACFP, the method or process comprising or consisting of the following steps:

[0133] mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP, and

[0134] The pH is lowered to below 7. Preferably, the pH is lowered to 6 or below 6, preferably 5.5. Typically, the pH is maintained below 7, more preferably the pH is maintained at 6 or below 6, even more preferably 5.5.

[0135] In any aspect, the step of mixing a solvent with a powder comprising or consisting of PP-stabilized ACP and / or ACFP comprises adding the solvent to the powder. Alternatively, the step comprises adding the powder to the solvent.

[0136] In any method or process for preparing a liquid composition as described herein, the method or process further comprises the step of degassing the liquid composition. Degassing can be performed by any method of creating a negative pressure above the liquid composition, including the methods described herein.

[0137] In any method or process for preparing a liquid composition as described herein, the method or process further comprises the step of mixing the liquid composition with a solution comprising fluoride ions.

[0138] In another aspect, the present invention provides a method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or PP-stabilized ACFP, the method or process comprising or consisting of the steps described in Example 1 herein.

[0139] In any aspect, the present invention provides a method or process further comprising the following steps to prepare a powder comprising or consisting of PP-stabilized ACP and / or ACFP:

[0140] mixing one or more solutions comprising phosphopeptides, calcium ions, phosphate ions, hydroxide ions, and optionally fluoride ions while maintaining the pH above about 7.0, preferably about 9, to form a solution comprising stabilized ACP and / or stabilized ACFP, and

[0141] drying a solution comprising PP-stabilized ACP and / or PP-stabilized ACFP,

[0142] Thereby a powder comprising or consisting of PP-stabilised ACP and / or ACFP is formed. Preferably, the drying is spray drying or freeze drying.

[0143] In one embodiment, the method or process further comprises the following steps:

[0144] The solution comprising the PP-stabilized ACP and / or the PP-stabilized ACFP is filtered prior to drying to form a retentate, wherein the retentate is subsequently dried to form a powder comprising or consisting of the PP-stabilized ACP and / or the PP-stabilized ACFP.

[0145] 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 PP-stabilized ACFP, the method or process comprising or consisting of the following steps:

[0146] mixing a solvent and a powder comprising or consisting of PP-stabilized ACP and / or PP-stabilized ACFP, and

[0147] The pH is lowered to below 8. Preferably, the solvent comprises a fluoride.

[0148] In this aspect, the method further comprises the step of stirring the liquid composition after lowering the pH.Preferably, stirring is performed for at least 5, 10, 15, 20, 25 or 30 minutes.

[0149] In this aspect, the liquid composition is degassed to remove trapped air bubbles, preferably by placing the solution under vacuum, most preferably for 24 hours.

[0150] 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 PP-stabilized ACP and / or PP-stabilized ACFP, the method or process comprising or consisting of the steps as described in Example 11 herein.

[0151] In any of the methods or processes for preparing a liquid composition comprising greater than 40% w / v or 40% w / w of stabilized ACP and / or stabilized ACFP, the solvent is water.

[0152] In any of the methods or processes for preparing a liquid composition comprising greater than 40% w / v or 40% w / w of stabilized ACP and / or stabilized ACFP, 1-10 M HCl or 11 M HCl is used to reduce or maintain the pH.

[0153] In any aspect, the method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or PP-stabilized ACFP can be used to prepare a liquid composition comprising equal to or greater than 45%, 50%, 55%, 60%, 65%, 70% or 75% w / v (or any other % w / v described herein) of stabilized ACP and / or ACFP.

[0154] In any method or process for preparing a liquid composition comprising at least 40% w / v of PP-stabilized ACP and / or PP-stabilized ACFP, the PP-stabilized ACP or PP-stabilized ACFP is a CPP-ACP or CPP-ACFP as described herein.

[0155] In any method or process for preparing a liquid composition comprising at least 40% w / v PP-stabilized ACP and / or PP-stabilized ACFP, the liquid composition is used in any dental treatment method, preferably those described herein (e.g., reducing the visibility of hypomineralized tooth surfaces or subsurfaces).

[0156] In another aspect, the present invention provides a liquid composition prepared by a method or process as described herein, comprising at least 40% w / v of PP-stabilized ACP and / or PP-stabilized ACFP.

[0157] The present invention also relates to a kit for treating or preventing one or more of caries, fluorosis, dental erosion and white spot lesions, comprising (a) a liquid composition comprising at least 40 wt% of amorphous calcium phosphate (ACP) stabilized by phosphopeptide (PP) and / or amorphous calcium phosphate fluoride (ACFP) stabilized by phosphopeptide (PP), the pH of the liquid composition being less than or equal to pH 6; and (b) another composition having an alkaline pH, preferably as described herein, wherein the other composition is applied to the tooth surface or subsurface after the liquid composition, and wherein the other (or second) composition causes the liquid (or first) composition to gel. Ideally, the kit also includes instructions for use for patients who need such tooth surface mineralization treatment, preferably for use in any of the methods described herein. The instructions for use may describe the use of the kit, which is used to treat or prevent one or more of caries, tooth decay, dental erosion, fluorosis and white spot lesions. In one embodiment, the liquid (or first) composition and the other (or second) composition are present in an amount suitable for treating the patient. Preferably, the phosphopeptide (defined below) is a casein phosphopeptide. Preferably, the PP-stabilized ACP or PP-stabilized ACFP is in the form of a casein phosphopeptide-stabilized ACP or ACFP complex.

[0158] In one embodiment, the present invention provides any kit as described herein for use in or when used in a cosmetic method to disguise or conceal the visual appearance of a hypomineralized surface or subsurface. Typically, the kit may also include written instructions for use in a cosmetic method as described herein.

[0159] The kit of the present invention may also include a free fluoride ion source. Examples of free fluoride ion sources include, but are not limited to, the following: sodium fluoride, stannous fluoride, silver fluoride, amine fluoride, or any metal ion fluoride salt. These fluoride ion sources can be provided in the form of a solution (usually an aqueous solution) or a suspension.

[0160] In one aspect, the present invention also provides a kit comprising or consisting of:

[0161] (a) a first composition comprising a powder of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP;

[0162] (b) a second composition comprising a fluoride solution having a pH less than or equal to pH 6; and

[0163] (c) A third composition having an alkaline pH.

[0164] Preferably, the alkaline pH of the third composition may be about 9, 10, 11, 12, 13 or 14. Preferably, the third composition has an alkaline pH of about 9 or greater. Preferably, the first composition comprises an amount of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP that, when mixed with the second composition, forms a liquid composition comprising at least 40% w / v of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP.

[0165] In one embodiment, the kit comprises or consists of:

[0166] (a) 5 g CPP-ACP and / or CPP-ACFP,

[0167] (b) 5 ml of 0.73 M NaF dissolved in 1.146 M HCl, and

[0168] (c) 1.5 M sodium hydroxide.

[0169] Preferably, the set further comprises two micro brushes.

[0170] Preferably, the kit further comprises written instructions for using the kit in any of the methods described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0171] Figure 1 : 45% w / v CPP-ACP (pH 5.5) solution followed by 1 M NaOH solution was used to mask the white spot lesions. The left side shows the treated lesions and the right side shows the control.

[0172] Figure 2 : A solution of 63% w / v CPP-ACP and 8,200 ppm F (as NaF) (pH 5.5, undegassed) followed by 1 M NaOH solution was used to mask white spot lesions. Left: treated lesions, right: control.

[0173] Figure 3: Using the dental kit described in Example 3, part (a) was added to part (b) and mixed thoroughly. The mixture was then applied to the white spot lesion on the left with a first microbrush (see image of the lesion before application in (A)). After a few seconds / minutes at 37°C, solution (c) was then applied to the white spot lesion on the left with a second microbrush. Within 15 minutes, a reaction occurred in the white spot to form a gel and hide the white spot (see image in (B)).

[0174] Figure 4 : A solution of 75% w / v CPP-ACP and 9,880 ppm F (as NaF) at pH 5.5, followed by 2M NaOH solution, was used to mask white spot lesions. Left: treated lesions, right: control.

[0175] Figure 5 : A mixed composition formed by a liquid composition of 63% w / v CPP-ACP and 8,000 ppm F (pH 5.5, degassed) and a 1.5 M NaOH solution was used to mask white spot lesions. The left side shows the treated lesion and the right side shows the control.

[0176] Figure 6 : Treatment of demineralized dentin with a mixed composition of a liquid composition of 63% w / v CPP-ACP and 8,000 ppm F (pH 5.5) and 1.5 M NaOH solution. Left: treated lesion, right: control.

[0177] FIG7 : (A and B) Representative transverse microradiographic images (TMR) showing the formation of a protective layer on demineralized dentin using a mixed composition formed of a liquid composition of 63% w / v CPP-ACP and 8,000 ppm F (pH 5.5) and a 1.5 M NaOH solution. This is a TMR image taken approximately 20 minutes after application of the mixed composition.

[0178] FIG8 : Representative scanning electron microscope (SEM) images showing the formation of a protective layer on demineralized dentin with exposed dentinal tubules using (A) a mixed composition of a liquid composition of 63% w / v CPP-ACP and 8,000 ppm F (pH 5.5) and 1.5 M NaOH solution, and (B) control dentin.

[0179] FIG. 9 : Representative scanning electron microscope (SEM) images showing (A) spontaneous cracking of the protective layer in FIG. 8 after dehydration, and (B) SEM-EDS elemental analysis of the protective layer.

[0180] Fig.10 : Microradiographic images showing the protective layer formed on root dentin treated with premixed CPP-ACFP and 2M NaOH (incubated at 37°C for 48 hours).

[0181] Fig.11 : Microradiographic images showing the protective layer formed on the etched enamel surface treated with premixed CPP-ACFP and 2M NaOH (incubated at 37°C for 48 hours).

[0182] Figure 12: Masking of white spot lesions using the composition of 60% w / w CPP-ACP and 7,800 ppm F (pH 7.8) (75% w / v CPP-ACP with 10,000 mg / LF at pH 7.8) described in Example 11. (a) Before treatment; (b) after 40 seconds of curing; (c) after 15 minutes of curing; and (d) after 23 hours of curing. DETAILED DESCRIPTION

[0183] Other aspects of the present invention and other embodiments of the aspects described in the preceding paragraphs will become apparent from the following description given by way of example and with reference to the accompanying drawings. It should be understood that the invention disclosed and defined in this specification can be extended to all alternative combinations of two or more individual features mentioned or apparent in the text or drawings. All these different combinations constitute various alternative aspects of the present invention.

[0184] Reference will now be made in detail to certain embodiments of the present invention. Although the present invention will be described in conjunction with the embodiments, it should be understood that it is not intended to limit the present invention to those embodiments. On the contrary, the present invention is intended to cover all substitutions, modifications and equivalents that may be included within the scope of the present invention defined by the claims.

[0185] One skilled in the art will recognize a variety of 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.

[0186] All patents and publications mentioned herein are incorporated by reference in their entirety.

[0187] For the purpose of interpreting this specification, terms used in the singular form also include the plural form and vice versa.

[0188] As used herein, unless the context requires otherwise, the term "includes" and variations of the term, such as "comprising", are not intended to exclude further additives, components, integers or steps. As used herein, unless the context requires otherwise, "includes" and "comprising" can be used interchangeably.

[0189] One aspect of the present invention is based on several surprising discoveries, the first of which is that compositions comprising high concentrations of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP) can be maintained in a liquid state (i.e., without forming a gel). Prior to the present invention, it was believed that high concentrations of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP would cause the composition to form a gel or paste, and all liquid compositions described to date have relatively low concentrations of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP. A second surprising discovery is that increasing the pH of an acidic liquid composition changes the form of the composition to a gel, which may occur within and / or on a tooth surface, subsurface, or lesion. Without being bound by any theory or mode of action, it is believed that increasing the pH destabilizes the PP-ACP and / or PP-ACFP complexes and forms an amorphous embryonic hydroxyapatite or fluorohydroxyapatite gel. A third surprising discovery is that upon raising the pH of the composition (e.g., by applying another composition having an alkaline pH), the formation of this gel occurs rapidly. A fourth surprising discovery is that the gel formed changes the refractive index and reduces the visibility of hypomineralized surfaces, subsurfaces, or lesions by restoring translucency to the hypomineralized surfaces, subsurfaces, or lesions. An advantage of the present invention is that it can mask or hide visible hypomineralized lesions within a few minutes of a single patient visit and dental procedure, resulting in a significant improvement in appearance and reduced visibility, i.e., restoration of translucency to the surface. In addition, it does the same for calcium and phosphate (with or without fluoride). This represents a significant improvement in current dental procedures for treating lesions such as white spots. While providing cosmetic benefits, the gel also provides a reservoir of highly concentrated calcium, phosphate, and optionally fluoride to remineralize the surface, subsurface, or lesion.

[0190] The pH of the liquid composition containing a high concentration of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP is generally 6 or less and can be combined or mixed with other liquid compositions of alkaline pH before being applied to the tooth surface. Another surprising discovery is that the formed combined or mixed composition (which contains a high concentration of phosphopeptide-stabilized ACP and / or phosphopeptide-stabilized ACFP, pH equal to or greater than about 9) can be maintained in a form that allows it to be applied to the tooth surface. In other words, the combined or mixed composition can exist in liquid form for a period of time, such as 1 to 2 minutes, so that it can be applied to the tooth surface.

[0191] Another surprising discovery is that compositions comprising high concentrations of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP) can remain liquid (i.e., do not form a gel) and remain liquid at near-neutral pH values. The compositions do not need to be prepared at the physical location where the individual is treated during a dental professional procedure. The liquid compositions can also be used to reduce the visibility of white spot lesions, reduce dentin hypersensitivity, and other uses as described herein.

[0192] The tooth subsurface is typically a hypomineralized lesion such that the first and second compositions, or the mixed composition, in contact with the tooth surface migrate through any surface layer, i.e., pellicle and / or plaque, through the porous tooth surface to the area requiring mineralization. Preferably, the PP-stabilized ACP or PP-stabilized ACFP is in the form of a casein phosphopeptide-stabilized ACP or ACFP complex. The tooth surface is preferably tooth enamel. The tooth surface may be a lesion in tooth enamel, such as a lesion caused by caries, tooth erosion, or fluorosis.

[0193] The reduction in visibility of a hypomineralized surface, subsurface, or lesion can be determined simply by visual inspection by the human eye. The reduction in visibility can be any degree of reduction such that the hypomineralized surface, subsurface, or lesion is no longer apparent. The reduction in visibility can cause the hypomineralized surface, subsurface, or lesion to appear translucent, thereby having little or no difference from the surrounding normally mineralized tooth surface as determined by the human eye.

[0194] The visibility of a surface, subsurface or lesion can also be determined as follows. A colorimeter (Minolta ChromaMeter CR241, Minolta, Japan) can be used to record surface reflectance. Surface reflectance measurement was established by the Commission de L'Eclairage in 1978 in the L*a*b* color gamut, measuring human color perception involving three color dimensions (Commission Internationale de L'Eclaige (1978). Recommendations on uniform colour spaces, colour difference equations and psychometric colour terms. Paris: Bureau Centrale de la DIE Suppl. 2:15.). The L* value represents the color gradient from white to black, the a* value represents the color gradient from green to red, and the b* value represents the color gradient from blue to yellow (Commission Internationale de L'Eclaige, 1978). Only L* value measurements can be used for white colors with higher readings and darker colors with lower readings. To ensure that the position of the sample in the colorimeter is repeatable, a wax mold can be prepared and stored for each sample. All samples were allowed to air dry for 60 seconds using a dental triple syringe before each measurement. A single sample was repositioned ten times before and after treatment and the color reflectance L* value was recorded.

[0195] Dentin hypersensitivity results when the protective enamel or cementum covering the dentin is lost. Cementum is generally more susceptible to breakdown than enamel because it is thinner and more easily attacked by acid. However, breakdown of cementum does not occur until the gums recede and the root surfaces are exposed to the oral environment. Patients with cementum breakdown and dentin hypersensitivity often experience pain when the exposed areas of the teeth are exposed to cold air, hot or cold liquids, sweet or acidic foods, or when they come into contact with metal objects. Patients with dental hypersensitivity have more open dentinal tubules and / or tubules with larger diameters than normal.

[0196] An advantage of one aspect of the present invention is the formation of a protective layer. This layer generally has the same or similar composition as hydroxyapatite or fluorapatite. It can be formed on enamel or dentin and can be used to seal or occlude dentinal tubules, thereby reducing dentin sensitivity.

[0197] Such a layer may be characterized as having a calcium:phosphate ratio equal to that of normal apatite, preferably a ratio of about 1.5 to 2: 1. The layer ideally contains about 20 wt% calcium.

[0198] Preferably, the layer comprises carbon, oxygen, phosphate and calcium, and optionally fluoride.

[0199] The method of sealing exposed dentin of the present invention reduces tooth sensitivity and reduces the risk of caries, such as root surface caries. In addition, since dental restoratives shrink and form micro gaps with the tooth surface, the present invention is particularly applicable before the application of restorative materials (such as compositions including glass ionomer cements). At this time, the gel or protective layer will act as a cavity sealant and reduce the formation of micro gaps.

[0200] The term "treat" or "treatment" refers to therapeutic treatment, the purpose of which is to slow down (mitigate) an undesirable physiological change or disorder. For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, relief of symptoms, relief of the extent of the condition, a stable (i.e., non-worsening) state of the condition, a delay or slowing of the progression of the condition, an improvement or alleviation of the disease / condition state, and relief (whether partial or complete), whether detectable or undetectable. Treatment may not necessarily result in the complete absence of detectable symptoms of the condition, but may reduce or minimize complications and side effects of the condition. The success of the treatment can be monitored by physical examination of the individual or by response to any thermal, tactile or chemical treatment as described herein. When the method of the present invention is used to treat dentin hypersensitivity or allergy in a subject, preferably, the subject experiences a decrease in the severity of pain or a decrease in the incidence of pain over time. Methods for identifying subjects with varying degrees of dentin sensitivity and for measuring treatment or prevention success are described herein and also include those outlined in Med Oral Patol Oral Cir Bucal.; 2008 Mar 1;13(3):E201-6. Treatment of a subject can be determined by comparing the level of pain experienced when exposed to any of the stimuli described herein before and after treatment, wherein a reduction in pain after treatment indicates a reduction in sensitivity.

[0201] The terms "prevent" and "prevention" generally refer to preventive or prophylactic measures used to protect or prevent individuals who do not have a condition or symptom (such as sensitivity) from developing a condition or symptom (such as sensitivity). Individuals who may need prevention are those who are undergoing dental procedures, particularly those that expose dentin.

[0202] In any aspect of the present invention, in (i), low mineralized tooth surface or subsurface or lesion is contacted with liquid composition for a period of time to allow liquid composition to penetrate tooth surface, subsurface or lesion. Typically, liquid composition is applied for a period of time to allow liquid composition to penetrate the pores of low mineralized tooth surface or subsurface or lesion. This provides liquid composition in those pores so that when the pH of the liquid composition in the pores increases to form gel, the visibility of low mineralized tooth surface or subsurface or lesion can be reduced. Therefore, gel can be formed in subsurface or lesion, or formed within and on subsurface or lesion. In one embodiment, before the pH of the liquid composition applied to low mineralized tooth surface or subsurface is increased to equal to or greater than about 9, low mineralized tooth surface or subsurface or lesion is contacted with liquid composition for up to 20 minutes in (i). In one embodiment, low mineralized tooth surface or subsurface or lesion is contacted with liquid composition for at least about a few seconds to about 5 minutes, preferably at least about 5 minutes to about 20 minutes in (i).

[0203] Typically, the gel may form any time up to about 5 to 20 minutes after the other composition is applied. Thus, in one embodiment, after the other composition is applied to raise the pH of the first composition, a period of about 5 to about 20 minutes is allowed to elapse before any other composition is applied or the step is applied to the tooth surface, subsurface or lesion.

[0204] As used herein, % w / v may be considered to be equivalent to g / 100 ml.

[0205] In any aspect, the tooth surface is in need of such treatment. Thus, in another aspect, in addition to the steps of any of the methods described herein, the invention comprises the step of identifying a subject suffering from fluorosis, caries, dentin hypersensitivity or calculus, white spot lesions, fluorosis lesions, carious lesions, or lesions caused by dental erosion.

[0206] Other compositions of alkaline pH are compositions with a pH greater than 7, which contain alkali or compounds capable of producing alkali. Alkali is defined as a compound that can accept hydrogen cations (protons) or (more generally) provide a pair of valence electrons. Compositions may include compounds that are not necessarily considered to be alkalis, such as polypeptides with many acidic and basic residues but still have the ability to increase the pH of the composition to greater than 7 (preferably to about pH 9 or more). Non-limiting examples of alkalis suitable for the present invention include hydroxides, borates, phosphates (including hydrogen phosphates), amines and any salt forms thereof (including alkali metal salt forms). More specifically, non-limiting examples of suitable pharmaceutically acceptable alkalis include ammonium hydroxide, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, ferrous hydroxide, zinc hydroxide, sodium hypochlorite, copper hydroxide, aluminum hydroxide, ferric hydroxide, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, lysine, arginine, histidine or urea. In one embodiment, the further composition comprises sodium hydroxide, preferably at a concentration greater than or equal to 1 or 2M.

[0207] Any pharmaceutically acceptable compound described as a base is suitable for the present invention. Generally, the base is suitable for oral use. Preferably, the compound behaves as a base, that is, only releases hydroxide ions or provides electrons in the presence of an acid. The base can be in the form of a free base or a pharmaceutically acceptable salt. Non-limiting examples of bases suitable for the present invention include hydroxides, borates, phosphates (including hydrogen phosphates and dihydrogen phosphates), citrates, carbonates, bicarbonates, hypochlorites, amines and any salt forms thereof (including alkali metal salt forms). More specifically, non-limiting examples of suitable pharmaceutically acceptable bases include ammonium hydroxide, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, ferrous hydroxide, zinc hydroxide, copper hydroxide, aluminum hydroxide, ferric hydroxide, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, lysine, arginine, histidine. Hypofluorite, which can act as a base as described herein, can also be used as a reagent to increase or maintain pH in the present invention. Suitable hypofluorites will react in situ to produce fluoride ions and hydroxide (or other base) ions. It will be appreciated by those skilled in the art that fluoride ions can replace hydroxide in the apatite crystal structure to form fluorapatite.

[0208] Any heat source can be used to heat or cure the tooth surface or subsurface in the method or use of the present invention. Heat sources that emit light or radiation and are suitable for dental applications are known in the art. Specific examples include dental curing lamps, such as 10W high-power blue light LEDs, such as X-Cure from Guilin Woodpecker Medical Instrument Co., Ltd. In any method or use of the present invention, after the liquid composition contacts the tooth surface or subsurface, there may be an additional step of heating or curing the liquid composition. Heating or curing can last for equal to or at least 30 seconds, equal to or at least 40 seconds, equal to or at least 50 seconds, equal to or at least 60 seconds, equal to or at least 2 minutes, or equal to or at least 5 minutes. Heating or curing may suddenly raise the temperature to 45-50°C (with patient comfort). Heating or curing can be carried out for any time or at any temperature as described herein (including embodiments).

[0209] Any composition described herein can be applied to a tooth surface, subsurface, or lesion using any technique known in the art or described herein. An exemplary application technique is using a microbrush.

[0210] Additionally, any resin barrier, such as a rubber dam, may be used to protect the soft tissue in the oral cavity from the application of any of the compositions described herein.

[0211] As used herein, "stable ACP or stable ACFP" and "stable ACP or stable ACFP complex" are used interchangeably.

[0212] The stable ACP or stable ACFP complexes described in the current specification may be "closed" complexes, such as those described in Cross et al. (2007). Figure 2 as shown in .

[0213] A stabilized ACP or stabilized ACFP complex as referred to herein includes a stabilized ACP or stabilized ACFP complex as described in WO2006 / 056013 (PCT / AU2005 / 001781), the contents of which are incorporated by reference.

[0214] In a preferred embodiment, the phosphopeptide-stabilized amorphous calcium phosphate (ACP) or phosphopeptide-stabilized amorphous calcium fluoride phosphate (ACFP) complex has tightly bound and loosely bound calcium, wherein the bound calcium in the complex is lower than the tightly bound calcium in the ACP or ACFP complex formed at pH 7.0. Optionally, the ACP or ACFP is primarily in alkaline form.

[0215] The stable ACP or stable ACFP complex referred to herein includes a stable ACP or stable ACFP complex formed at a pH value below 7.0. Preferably, the complex is formed in a pH range of about 5.0 to below 7.0. More preferably, the complex is formed in a pH range of about 5.0 to about 6.0. In a preferred embodiment, the complex is formed at a pH value of about 5.0 or about 5.5. Preferably, the ACP or ACFP in the complex is mainly in an alkaline form.

[0216] Stable ACP can be produced by a method comprising the following steps:

[0217] (i) obtaining a solution comprising at least one phosphopeptide, and

[0218] (ii) mixing a solution comprising calcium ions, phosphate ions and hydroxide ions while maintaining a pH value of about 5.5 to 9.

[0219] In one embodiment, the pH is maintained below 7.0.

[0220] Stable ACFP can be produced by a method comprising the following steps:

[0221] (i) obtaining a solution comprising at least one phosphopeptide, and

[0222] (ii) mixing a solution comprising calcium ions, phosphate ions, hydroxide ions and fluoride ions while maintaining a pH value of about 5.5 to 9.

[0223] In one embodiment, the pH is maintained below 7.0.

[0224] Hydroxyl ions can be titrated into the solution to maintain the phosphopeptide solution at a substantially constant pH. Calcium ions and phosphate ions can be titrated into the phosphopeptide solution with constant stirring and at a rate that avoids the formation of calcium phosphate precipitates in the phosphopeptide solution.

[0225] The phosphopeptide-stabilized amorphous calcium phosphate (ACP) or phosphopeptide-stabilized amorphous calcium fluoride phosphate (ACFP) complex may also include a complex in which the ACP in the complex has tightly bound calcium and loosely bound calcium, wherein the tightly bound calcium in the complex is less than the tightly bound calcium in the ACP or ACFP complex formed at a pH of 7.0, and the ACP or ACFP is predominantly in an alkaline form, obtainable by a method comprising the steps of:

[0226] a) mixing a first solution comprising calcium ions, a second solution comprising phosphate ions, and optionally a third solution comprising fluoride ions into a solution comprising the phosphopeptide and a solvent having a pH of about 5 to below 7; and

[0227] b) The pH of the solution is maintained at about 5.0 to below 7.0 by adding hydroxide ions during mixing.

[0228] "Tightly" and "loosely" bound calcium and phosphate can be determined using ultrafiltration analysis. Briefly, a solution of mixed phosphopeptides, calcium, phosphate, and optional fluoride while maintaining the pH at about 7.0 or below can first be filtered through a 0.1 micron filter to remove free calcium and phosphate that are not bound to the complex. This free calcium and phosphate are present in the filtrate and are discarded. Any free calcium or phosphate that is not bound to the complex in any way is not bioavailable, i.e., delivered to the teeth by the phosphopeptides. The retentate from the 0.1 micron filtration can be further analyzed by centrifugation through a 3000 mw cutoff filter at a speed of 1,000 g for 15 minutes. The resulting filtrate contains calcium and phosphate that are loosely bound or associated with the complex. Under this centrifugal force, calcium and phosphate that are not tightly bound to the complex are released and move into the filtrate. Ca and Pi tightly bound to the complex 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 of the 0.1 micron filtration.

[0229] The stabilized ACP or stabilized ACFP complex referred to herein includes the stabilized ACP or stabilized ACFP complex as described in WO2006 / 135982 (PCT / AU2006 / 000885), the contents of which are incorporated by reference.

[0230] Amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex stabilized by a "superloaded" phosphopeptide or phosphoprotein (PP). The complex can be formed at any pH value (e.g., 3-10). Preferably, the phosphopeptide comprises the sequence -ABC-, wherein A is a phosphoamino acid, preferably phosphoserine, B is any amino acid, including a phosphoamino acid, and C is glutamic acid, aspartic acid, or a phosphoamino acid. The phosphoamino acid can be phosphoserine. PP is superloaded with calcium ions and phosphate ions. The calcium ion can be 30-1000 moles of Ca / mole of PP, or 30-100 or 30-50 moles of Ca / mole of PP. In another embodiment, the moles of Ca / mole of PP are at least 25, 30, 35, 40, 45, or 50.

[0231] The present invention comprises amorphous calcium phosphate or amorphous fluorinated calcium phosphate complexes stabilized with phosphopeptides or phosphoproteins (PP) having 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.

[0232] The present invention also provides a phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex, which is produced by a method comprising the following steps:

[0233] (i) obtaining a solution comprising calcium, inorganic phosphate and, optionally, fluoride; and

[0234] (ii) mixing (i) with a solution comprising PP-ACP.

[0235] In a preferred embodiment, the PP is casein phosphopeptide (CPP).

[0236] In another aspect, the present invention also includes the use of a formulation of a PP-stabilized ACP and / or a PP-stabilized ACFP complex together with at least an equal weight of calcium phosphate. Preferably, the calcium phosphate is CaHPO4 or calcium lactate or any other soluble calcium phosphate compound. Preferably, the calcium phosphate (e.g., CaHPO4) is dry-mixed with the PP-stabilized ACP and / or the PP-stabilized ACFP complex. In a preferred embodiment, the ratio of PP-ACP and / or PP-ACFP complex: calcium phosphate is about 1:1-50, more preferably about 1:1-25, more preferably about 1:5-15. In one embodiment, the ratio of PP-ACP and / or PP-ACFP complex: calcium phosphate is about 1:10.

[0237] An oral care formulation comprising a phosphopeptide- or phosphoprotein (PP)-stabilized amorphous calcium phosphate (ACP) and / or a phosphopeptide- or phosphoprotein-stabilized amorphous calcium phosphate (ACFP) complex has a calcium ion content greater than about 30 moles of calcium per mole of PP when used in the oral cavity and can be made by a process comprising the steps of:

[0238] (i) obtaining a powder comprising a PP-ACP and / or PP-ACFP complex;

[0239] (ii) dry mixed with an effective amount of calcium phosphate; and

[0240] (iii) formulating the dry-blended PP-ACP and / or PP-ACFP and calcium phosphate mixture into an oral care formulation.

[0241] Preferably, the form of calcium phosphate used for dry mixing is any soluble calcium phosphate, including but not limited to CaHPO4, Ca2HPO4 and calcium lactate.

[0242] Compositions as described herein may also contain free fluoride ions. Fluoride ions may come from any suitable source. Fluoride ion sources may include free fluoride ions or fluoride salts. Examples of fluoride ion sources include, but are not limited to, the following: sodium fluoride, sodium monofluorophosphate, stannous fluoride, sodium fluorosilicate, and amine fluorides. These may be provided in the form of a solution (usually an aqueous solution) or a suspension.

[0243] 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, it is preferably greater than 10 ppm. In the typical embodiments described below, the amount can be hundreds or thousands of ppm. The fluoride content is typically measured in ppm in the oral composition in a manner commonly used in the art. If the fluoride is provided from a source containing stable ACP, the ppm refers to the concentration of fluoride in the source, typically a solution or suspension of bioavailable fluoride.

[0244] The stannous associated ACP or ACFP complexes referred to herein include any of those described in PCT / AU2014 / 050447, the entire contents of which are incorporated by reference in their entirety.

[0245] The composition as described herein for use in the method of use of the present invention may include a stannous associated ACP or ACFP complex. The composition may include 2% CPP-ACP and 290 ppm fluoride, of which 220 ppm is as stannous fluoride and 70 ppm is as sodium fluoride.

[0246] In any aspect or embodiment described herein, the stabilized ACP and / or stabilized ACFP is stabilized by a phosphopeptide (PP). 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.

[0247] "Phosphopeptide" in the present specification refers to an amino acid sequence in which at least one amino acid is phosphorylated. Preferably, the phosphopeptide includes one or more amino acid sequences -ABC-, wherein A is a phosphoamino residue, B is any aminoacyl residue including a phosphoamino residue, and C is selected from a glutamyl residue, an aspartyl residue or a phosphoamino residue. Any phosphoamino residue can be independently a phosphoseryl residue. B is ideally a residue whose side chain is neither relatively large nor hydrophobic. It can be Gly, Ala, Val, Met, Leu, Ile, Ser, Thr, Cys, Asp, Glu, Asn, Gln or Lys. Preferably, at least two phosphoamino acids in the sequence are preferably continuous. Preferably, the phosphopeptide includes the sequence ABCDE, wherein A, B, C, D and E are independently phosphoserine, phosphothreonine, phosphotyrosine, phosphothistidine, 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 residue is phosphoserine, most preferably three consecutive phosphoserine residues. It is also preferred that D and E are independently glutamic acid or aspartic acid.

[0248] In one embodiment, the ACP or ACFP is stabilized by a casein phosphopeptide (CPP), which is in the form of intact casein or a casein fragment, and the complex formed preferably has the formula [CPP (ACP) 8] n or [(CPP)(ACFP)8] n , wherein n is equal to or greater than 1, such as 6. The complex formed may be a colloidal complex, wherein the core particles aggregate to form large (eg 100 nm) colloidal particles suspended in water. Thus, the PP may be casein or a phosphopeptide.

[0249] PP may be from any source; it may be present in a larger polypeptide, including a full-length casein polypeptide, or it may be isolated by trypsin or other enzymatic or chemical digestion of casein or other phosphoamino acid-rich proteins such as phosphitin, or by chemical or recombinant synthesis, provided that it contains the sequence -ABC- or ABCDE as described above. The sequences flanking this core sequence may be any sequence. However, α s1 (59-79), β(1-25), α s2 (46-70) and α s2 Those flanking sequences in (1-21) are preferred. The flanking sequences may optionally be altered by deletion, addition or conservative substitution of one or more residues. The amino acid composition and sequence of the flanking regions are not critical.

[0250] The phosphopeptide may be selected from any of those described in WO2006 / 056013, WO2006 / 135982 or US Pat. No. 5,015,628.

[0251] Examples of conservative substitutions are shown in Table 1 below.

[0252] Table 1

[0253] Original residue Exemplary conservative substitutions Conservative substitutions are preferred Ala Val,Leu,Ile Val Asn Gln Lys His Phe Gln Gln Asn Asn Gly Pro Pro Ile Leu,Val,Met,Ala,Phe Leu Leu Ile,Val,Met,Ala,Phe Ile Lys Arg,Gln,Asn Arg Phe Leu,Val,Ile,Ala Leu Pro Gly Gly Ser Thr Thr Val Ile,Leu,Met,Phe,Ala Leu Asp Glu Glu Thr Ser Ser Trp Tyr Tyr Tyr Trp Phe Thr Ser Phe

[0254] Flanking sequences may also include non-naturally occurring amino acid residues. Common amino acids not encoded by the genetic code include:

[0255] 2-aminoadipic acid (Aad) for Glu and Asp;

[0256] 2-aminopimelic acid (Apm) for Glu and Asp;

[0257] 2-aminobutyric acid (Abu) for Met, Leu, and other aliphatic amino acids;

[0258] 2-aminoheptanoic acid (Ahe) for Met, Leu, and other aliphatic amino acids;

[0259] 2-aminoisobutyric acid (Aib) for Gly;

[0260] Cyclohexylalanine (Cha) for Val, Leu, and Ile;

[0261] homoarginine (Har) for Arg and Lys;

[0262] 2,3-diaminopropionic acid (Dpr) for Lys, Arg, and His;

[0263] N-ethylglycine (EtGly) for Gly, Pro, and Ala;

[0264] N-ethylasparagine (EtAsn) for Asn and Gln;

[0265] Hydroxylysine (Hyl), for Lys;

[0266] allohydroxyllysine (AHyl), for Lys;

[0267] 3-(and 4) hydroxyproline (3Hyp, 4Hyp) for Pro, Ser, and Thr;

[0268] Alloisoleucine (Alle), for Ile, Leu, and Val;

[0269] ρ-amidinophenylalanine for Ala;

[0270] N-methylglycine (MeGly, sarcosine), for Gly, Pro, Ala.

[0271] N-methylisoleucine (MeIle) for Ile;

[0272] Norvaline (Nva), for Met and other aliphatic amino acids;

[0273] norleucine (Nle), for Met and other aliphatic amino acids;

[0274] Ornithine (Orn) for Lys, Arg, and His;

[0275] citrulline (Cit) and methionine sulfoxide (MSO) for Thr, Asn, and Gln;

[0276] N-methylphenylalanine (MePhe), trimethylphenylalanine, halogen (F, Cl, Br and I) phenylalanine, trifluorophenylalanine, for Phe.

[0277] In one embodiment, PP is one or more selected from α s1 (59-79)[1], β(1-25)[2], α s2 (46-70)[3] and α s2 (1-21)[4] phosphopeptide:

[0278] [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-Lys79(SEQ ID NO:1)α s1 (59-79)

[0279] [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)

[0280] [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)

[0281] [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).

[0282] In certain preferred forms of the invention, the liquid composition may be a mouthwash, rinse or spray. In such formulations, the carrier is typically a water-alcohol mixture, desirably including a wetting agent. Typically, the weight ratio of water to alcohol is in the range of about 1:1 to about 20:1. In this type of formulation, the total amount of the water-alcohol mixture is typically in the range of about 70 to about 99.9% by weight of the formulation. The alcohol is typically ethanol or isopropanol. Ethanol is preferred.

[0283] It will be appreciated that, generally speaking, oral preparations are usually sold or otherwise dispensed in appropriately labeled packaging. Thus, a bottle of mouthwash will have a label that essentially describes it as a mouth rinse or mouthwash, and instructions for use.

[0284] Before adding the liquid composition, a preparatory composition can be used to prepare (e.g., clean) the tooth surface, subsurface, or lesion. Such a composition may include the following components. An organic surfactant may be used in the composition to achieve an enhanced preventive effect, help achieve thorough and complete dispersion of the active agent throughout the oral cavity, and make the present composition more cosmetically acceptable. The nature of the organic surfactant is preferably anionic, nonionic, or amphoteric, and preferably does not interact with the active agent. Detergent materials that impart detergency and foaming properties to the composition are preferably used as surfactants. Suitable examples of anionic surfactants are water-soluble salts of higher fatty acid monoglycerides: monosulfates, such as sodium salts of monosulfated monoglycerides of hydrogenated coconut oil fatty acids; higher alkyl sulfates, such as sodium dodecyl sulfate; alkyl aryl sulfonates, such as sodium dodecylbenzene sulfonic acid; higher alkyl sulfoacetates; higher fatty acid esters of 1,2-dihydroxypropane sulfonic acid, and substantially saturated higher fatty acyl amides of lower aliphatic aminocarboxylic acid compounds, such as those having 12 to 16 carbon atoms in fatty acids, alkyl or acyl radicals, etc. Examples of the last mentioned amides are N-lauroylsarcosine, and the sodium, potassium and ethanolamine salts of N-lauroylsarcosine, N-myristoylsarcosine or N-palmitoylsarcosine, which should be substantially free of soaps or similar higher fatty acid materials. The use of these sarconite compounds in the oral compositions of the present invention is particularly advantageous because, in addition to reducing the solubility of tooth enamel in acidic solutions, these materials exhibit a significant and prolonged effect in inhibiting acid formation in the oral cavity caused by carbohydrate decomposition. Examples of suitable water-soluble nonionic surfactants are the condensation products of ethylene oxide with various reactive hydrogen-containing compounds having long hydrophobic chains (e.g., fatty chains of about 12 to 20 carbon atoms), the condensation products ("ethoxamers") containing hydrophilic polyoxyethylene moieties, such as the condensation products of poly(ethylene oxide) with fatty acids, fatty alcohols, fatty amides, polyols (e.g., sorbitan monostearate) and polypropylene oxide (e.g., Pluronic materials).

[0285] Various other materials may be added to 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. When present, these adjuvants are incorporated into the preparations in amounts that do not substantially adversely affect the desired properties and characteristics.

[0286] Any suitable flavoring or sweetening material may also be used. Examples of suitable flavoring ingredients are flavoring oils, for example, oils of spearmint, mint, holly, sassafras, cloves, sage, eucalyptus, marjoram, cinnamon, lemon and orange, and methyl salicylate. Suitable sweeteners include sucrose, lactose, maltose, sorbitol, xylitol, sodium cyclamate, perilla, AMP (aspartame), saccharin, etc. Suitably, flavoring agents and sweeteners may constitute about 0.1% to more than 5% of the preparation individually or together.

[0287] Other compositions at alkaline pH may further comprise other components to enhance gel formation, for example, the addition of stannous, zinc, magnesium or other metal ions or other chemicals that aid in cross-linking the phosphopeptide-stabilized ACP or phosphopeptide-stabilized ACFP to enhance gelation.

[0288] It should be clearly understood that although the present description specifically relates to applications in humans, the present invention can also be used for veterinary purposes. Thus, in all aspects, the present invention can be used for domestic animals, such as cattle, sheep, horses and poultry; for companion animals, such as cats and dogs; and for zoo animals.

[0289] The invention will now be further described with reference to the following non-limiting examples.

[0290] Example 1

[0291] Preparation of high concentration liquid CPP-ACFP and CPP-ACP solutions

[0292] 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 digest of casein until just before precipitation or gelation (usually yielding approximately 78 mM to 124 mM CaCl2). 2+and a final concentration of 48mM to 76mM inorganic phosphate). The solution is added slowly (i.e., less than about 1% volume per minute) with good stirring. First, add one portion of phosphate solution, then one portion of calcium solution. Use 1 to 10M NaOH to mix well and maintain the pH of the bulk solution at 9.0. Sodium hydroxide solution is added automatically by pH stat, and hydroxide ions are usually added after each addition of calcium ions. After the addition of calcium ions, phosphate ions, and hydroxide ions is completed, the solution is filtered through a 0.1 micron filter to concentrate 1-2 times. The retentate is then washed with 1-2 volumes of water to remove salts and inactive (and bitter) peptides. The prepared CPP-ACP solution is then spray dried or freeze dried to produce a white powder. The dry powder was then added to water to form a 45% w / v CPP-ACP solution at pH 5.5 by adding 1-10 M HCl or a 63% w / v CPP-ACP solution at pH 5.5 by adding 1-10 M HCl (with NaF added to produce 8,200 ppm F).

[0293] The 75% w / v solution was prepared by adding 75 g of CPP-ACP powder to 20 ml of water, adding small amounts of powder each time (0.5 g / min) while maintaining the pH at 5.5 by adding 10 M HCl. The solution was thoroughly mixed after each addition to ensure dispersion. Concentrated NaF (0.95 M) solution was added with 10 M HCl to ensure that 52 mmol of F was finally added. CPP-ACP powder, NaF and HCl were added with water and fixed to 100 ml over 2-3 hours. This produced a liquid composition of 75% w / v CPP-ACP, 9,880 ppm F, pH 5.5.

[0294] Example 2

[0295] Masking of white spot lesions using CPP-ACP and alkaline solutions

[0296] A 45% w / v CPP-ACP, pH 5.5 liquid solution was applied (within a few seconds) to the surface of the enamel blocks with white spot lesions using a microbrush. The blocks were then incubated at 37°C for 20 minutes. A 1 M NaOH solution (approximately pH 14) was then applied using a microbrush (within a few seconds) and the blocks were then incubated at 37°C for another 20 minutes.

[0297] Figure 1 A comparison of a treated white spot lesion (T) and a control lesion (C) is shown, which are from the same lesion (i.e., an enamel block cut in half). Compared to the control sample, in which the white spot lesion is still clearly visible, the white spot lesion in the treated sample is substantially masked by the treatment by restoring translucency to the lesion.

[0298] Example 3

[0299] Masking of leukoplakia lesions using 63% w / v CPP-ACP, free fluoride, and alkaline solutions

[0300] A liquid solution of 63% w / v CPP-ACP and 8,200 ppm F (as NaF), pH 5.5, was applied (within seconds) to the surface of the enamel blocks with white spot lesions using a microbrush. The blocks were then incubated at 37° C. for 20 minutes. A 1 M NaOH solution was then applied using a microbrush (within seconds) and the blocks were then incubated at 37° C. for another 20 minutes.

[0301] Figure 2 A comparison of a treated white spot lesion (T) on the left and a control lesion (C) on the right, from the same lesion (i.e., an enamel block cut in half), is shown. The white spot lesion in the treated sample is substantially masked by the treatment by restoring translucency to the lesion, compared to the control sample, in which the white spot lesion is still clearly visible.

[0302] Example 4

[0303] In the dental office, patients who need to reduce the visibility of hypomineralized tooth surfaces or subsurfaces of enamel can be treated using the following steps:

[0304] 1. Use a rubber dam and etch the leukoplakia [this step is optional and not necessary for very porous (active) lesions]. This may involve a standard liquid rubber dam (a resin barrier to protect the soft tissue) and an acid etching technique (e.g., 30% phosphoric acid or 15% HCl).

[0305] 2. Use a microbrush to apply a high concentration (>40% w / v) of stable CPP-ACP or stable CPP-ACP / F acid solution (e.g., below pH 6) to the white spots and then keep it for 5-20 minutes. Optionally, a high-intensity LED curing light (e.g., 10W high-power blue light) can be used to heat to suddenly increase the temperature to 45-50°C (within the patient's comfort).

[0306] 3. Apply high concentration alkali (e.g. 4% w / v NaOH) using a micro brush and hold for 5-20 minutes. Optionally, heat can be applied using a high intensity LED curing light (e.g. 10W high power blue light) to suddenly increase the temperature to 45-50°C (within patient comfort).

[0307] Example 5

[0308] Exemplary Dental Kits

[0309] The dental kit includes or consists of the following three parts:

[0310] (a) 5 g CPP-ACP powder, preferably prepared as described herein,

[0311] (b) 5 ml 0.73 M NaF dissolved in 1.146 M HCl, and

[0312] (c) 1.5 M NaOH, including two microbrushes.

[0313] Add part (a) to part (b) and stir thoroughly. Then apply this mixture to the white spot lesion on the left with the first micro brush (see Figure 3 (A) for an image of the lesion before application). After a few seconds / minutes at 37°C, solution (c) is then applied to the white spot lesion on the left with the second micro brush. Within 15 minutes, a reaction occurs in the white spot and a gel is formed and hides the white spot (see the image in Figure 3 (B)).

[0314] Example 6

[0315] Masking of leukoplakia lesions using 75% w / v CPP-ACP, free fluoride, and alkaline solutions

[0316] A liquid of 75% w / v CPP-ACP, 9,880 ppm F as NaF, pH 5.5 was applied with a microbrush (within seconds) to the surface of the enamel blocks with white spot lesions. The blocks were then incubated at 37°C for 10 minutes. A 2M NaOH solution was then applied with a microbrush (within seconds) and the blocks were then incubated at 37°C for another 20 minutes.

[0317] Figure 4 A comparison of a treated white spot lesion on the left and a control lesion on the right, from the same lesion (i.e., an enamel block cut in half), is shown. The white spot lesions in the treated samples were substantially masked by the treatment by restoring translucency to the lesions, compared to the control samples, where the white spot lesions were still clearly visible.

[0318] Example 7

[0319] Masking of white spot lesions by forming a mixed composition prior to application to the tooth surface

[0320] Prior to application to the tooth surface, a degassed 63% w / v CPP-ACP / 8,000 ppm F liquid composition at pH 5.5 was mixed with a 1.5 M NaOH solution. The mixed composition was then applied to the white spot lesions. A dental curing light at setting 2 was then applied to the surface for 40 seconds. Significant enamel white spot coverage ( Figure 5 ).

[0321] Example 8

[0322] Forms a protective layer on dentin

[0323] Another application of the present invention is to seal or occlude exposed root (dentin) surfaces (the aging population has exposed root surfaces and they are more susceptible to caries / erosion). To simulate these exposed root surfaces, the root dentin was treated with 15% EDTA for 2 minutes to remove the smear layer to expose the dentinal tubules ( Figure 6 , right block). The mixed composition as described in Example 7 was applied to the dentin to form a gel on the surface ( Figure 6 , left block).

[0324] Furthermore, when the mixed composition is applied to a dentin surface (whether sound or previously demineralized with an acidic buffer), the solution not only gels, but also begins to form a fluorapatite (FA) layer on the surface to seal the dentin (as shown in Figures 7, 8 and 9). This will have a significant impact on the reduction of tooth sensitivity and the risk of root caries. Interestingly, the fluorapatite layer is white, thus hiding the yellow dentin, thereby not only providing a seal and protective layer to reduce sensitivity and caries / erosion risk, but also improving the aesthetics of the tooth.

[0325] The protective layer formed was intentionally dehydrated (as shown in FIG. 9 ) to demonstrate its uniformity on the surface completely sealing the dentinal tubules (as shown in FIG. 8 on the right).

[0326] Elemental analysis showed that the formed protective layer had a composition similar to that of fluorohydroxyapatite ( Fig. 9B ).

[0327] Example 9

[0328] Forms a protective layer on dentin

[0329] method:

[0330] All dentin blocks were polished

[0331] Dentin blocks were treated with 15% EDTA for 2 minutes

[0332] 15 seconds etching (37% phosphoric acid)

[0333] Mix degassed 63% (w / v) CPP-ACP / 8100ppm F (pH 5.5) with an equal volume of 2M NaOH

[0334] Use a microbrush to topically apply a premixed solution of 63% (w / v) CPP-ACP / 8100 ppm F (pH 5.5) and 2 M NaOH to the dentin surface.

[0335] ·40 seconds light curing

[0336] Keep at 37℃ for 48 hours

[0337] · Section, grind and microradiograph.

[0338] like Fig.10 As shown, a mineral layer is formed on the surface of dentin.

[0339] Example 10

[0340] Forms a protective layer on tooth enamel

[0341] method:

[0342] All enamel blocks are polished

[0343] 15 seconds etching (37% phosphoric acid)

[0344] Mix 63% (w / v) CPP-ACP / 8100ppm F (pH 5.5) with an equal volume of 2M NaOH

[0345] Topically apply premixed 63% (w / v) CPP-ACP / 8100 ppm F (pH 5.5) and 2M NaOH to the enamel surface using a microbrush

[0346] ·40 seconds light curing

[0347] Keep at 37℃ for 48 hours

[0348] · Section, grind and microradiograph.

[0349] like Fig.11 As shown, a mineral layer forms on the surface of the enamel.

[0350] Embodiment 11

[0351] Masking of white spot lesions by forming a mixed composition prior to application to the tooth surface

[0352] 30 g of CPP-ACP powder (commercially available from Recaldent) was added to 19.5 g of a 20,000 ppm F (NaF) solution, to which was then added 0.5 g of a 11 M HCl solution to obtain a final weight of 50 g (the final solution was therefore a 60% w / w CPP-ACP, pH 7.8, solution containing 7,800 ppm F, or a pH 7.8, 10,000 mg / LF, 75% w / v CPP-ACP solution).

[0353] With thorough stirring (about 30 minutes), a homogeneous, very viscous but stable solution with a pH of 7.8 was prepared. The solution was then degassed by placing it under vacuum for 24 hours to remove trapped air bubbles.

[0354] The solution was applied to the white spot lesions using a microbrush and then light cured for 40 seconds using a dental curing light (as described above). Images before and after curing are shown (Figure 12(a) before; Figure 12(b) after curing for 40 seconds; Figure 12(c) after curing for 15 minutes; Figure 12(d) after curing for 24 hours). The cured stable high concentration solution was very effective in hiding the white spots, and the effect could be maintained for at least 24 hours at 37°C.

[0355] The viscous, stable and safe (neutral pH) solution is easy to apply in the dental procedure and the solution is more concentrated, thus producing better results for a longer period of time.

[0356] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more individual features mentioned or evident from the text or drawings. All these different combinations constitute various alternative aspects of the invention. Sequence Listing <110> The University of Melbourne <120> Hiddenness of hypomineralized lesions <130> 53065312AZD <150> AU2019900833 <151> 2019-03-13 <150> AU2019903860 <151> 2019-10-14 <160> 4 <170> PatentIn Version 3.5 <210> 1 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> alphas1(59-79) phosphopeptide <220> <221> MOD_RES <222> (6) <223> Phosphorylation <220> <221> MOD_RES <222> (8)..(10) <223> Phosphorylation <220> <221> MOD_RES <222> (17) <223> Phosphorylation <400> 1 Gln Met Glu Ala Glu Ser Ile Ser Ser Ser Glu Glu Ile Val Pro Asn 1 5 10 15 Ser Val Glu Gln Lys 20 <210> 2 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> beta(1-25) phosphopeptide <220> <221> MOD_RES <222> (15) <223> Phosphorylation <220> <221> MOD_RES <222> (17)..(19) <223> Phosphorylation <400> 2 Arg Glu Leu Glu Glu Leu Asn Val Pro Gly Glu Ile Val Glu Ser Leu 1 5 10 15 Ser Ser Ser Glu Glu Ser Ile Thr Arg 20 25 <210> 3 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> alphas2(46-70) phosphopeptide <220> <221> MOD_RES <222> (11)..(13) <223> Phosphorylation <220> <221> MOD_RES <222> (16) <223> Phosphorylation <400> 3 Asn Ala Asn Glu Glu Glu Tyr Ser Ile Gly Ser Ser Ser Glu Glu Ser 1 5 10 15 Ala Glu Val Ala Thr Glu Glu Val Lys 20 25 <210> 4 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Alphas2(1-21) phosphopeptide <220> <221> MOD_RES <222> (8)..(10) <223> Phosphorylation <220> <221> MOD_RES <222> (16) <223> Phosphorylation <400> 4 Lys Asn Thr Met Glu His Val Ser Ser Ser Glu Glu Ser Ile Ile Ser 1 5 10 15 Gln Glu Thr Tyr Lys 20

Claims

1. Use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for reducing the visibility of a hypomineralized tooth surface or subsurface, the liquid composition comprising at least 40% w / w and less than 80% w / w of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9.

2. Use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for treating or preventing dentin hypersensitivity in a subject in need thereof, the liquid composition comprising at least 40% w / w and less than 80% w / w of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being greater than or equal to pH 5 but less than or equal to pH 9.

3. The use according to claim 1, wherein the product is a cosmetic product.

4. The use according to claim 1, wherein The pH of the liquid composition is greater than or equal to 6, but less than or equal to 8.

5. The use according to claim 1, wherein The pH of the liquid composition is greater than or equal to 7, but less than or equal to 8.

6. The use according to claim 1, wherein The liquid composition also contains free fluoride ions.

7. The use according to claim 6, wherein The free fluoride ions are present in the liquid composition at a concentration of 200 ppm to 10000 ppm.

8. The use according to claim 6, wherein The free fluoride ions are present in the liquid composition at a concentration of 2600 ppm to 8500 ppm.

9. The use according to claim 6, wherein The free fluoride ions were present in the liquid composition at a concentration of 7800 ppm.

10. The use according to claim 1, wherein The use also includes heating the tooth surface or sub-surface or lesion.

11. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than 37°C.

12. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than or equal to 40°C.

13. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than or equal to 45°C.

14. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than or equal to 50°C.

15. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than or equal to 55°C.

16. The use according to claim 10, wherein The tooth surface or subsurface is heated to a temperature greater than or equal to 60°C.

17. The use according to claim 10, wherein: The tooth surface or subsurface is heated to a temperature greater than or equal to 65°C.

18. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 45% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

19. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 50% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

20. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 55% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

21. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 60% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

22. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 65% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

23. The use according to any one of claims 1 to 17, wherein The liquid composition comprises greater than or equal to 70% w / w and less than 80% w / w of stabilized ACP and / or stabilized ACFP.

24. Use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for reducing the visibility of a hypomineralized tooth surface or subsurface, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, wherein the liquid composition is applied to a hypomineralized tooth surface or subsurface, and the pH of the liquid composition applied to the hypomineralized tooth surface or subsurface is subsequently raised to equal to or greater than 9, thereby forming a gel in and / or on the hypomineralized tooth surface or subsurface.

25. Use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for forming a gel in and / or on a tooth surface or subsurface lesion, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, wherein the liquid composition is applied to a tooth surface or subsurface lesion, and then the pH of the liquid composition applied to the tooth surface or subsurface lesion is raised to equal to or greater than 9, thereby forming a gel in and / or on the tooth surface or subsurface lesion.

26. Use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for treating or preventing dentin hypersensitivity in a subject in need thereof, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, wherein the liquid composition is applied to a tooth surface or subsurface lesion, and the pH of the liquid composition applied to the tooth surface or subsurface lesion is subsequently raised to equal to or greater than 9, thereby forming a gel in and / or on the exposed dentinal tubules.

27. Use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP) in the manufacture of a product, the product comprising a liquid composition for forming a protective layer on a tooth surface, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, wherein the liquid composition is applied to a tooth surface or subsurface lesion, and then the pH value of the liquid composition applied to the tooth surface or subsurface lesion is raised to equal to or greater than 9.

28. Use according to any one of claims 24 to 27, wherein the product is a cosmetic product.

29. The use according to any one of claims 24 to 27, wherein The pH of the liquid composition applied to the tooth surface or subsurface lesion is raised to a pH of 10 or greater.

30. The use according to any one of claims 24 to 27, wherein The pH of the liquid composition applied to the tooth surface or subsurface is raised by contacting the liquid composition applied to the tooth surface or subsurface with another composition having an alkaline pH.

31. The use according to any one of claims 24 to 27, wherein The tooth surface is tooth enamel.

32. The use according to any one of claims 24 to 27, wherein The tooth surface is a lesion caused in the tooth enamel by caries, tooth erosion or fluorosis.

33. The use according to claim 32, wherein The lesions are leukoplakia lesions.

34. Use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for reducing the visibility of a hypomineralized tooth surface or subsurface, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, The liquid composition is mixed with another composition having an alkaline pH to form a mixed composition having a pH equal to or greater than 9, and the mixed composition is subsequently applied to a hypomineralized tooth surface or subsurface to form a gel in and / or on the hypomineralized tooth surface or subsurface.

35. Use of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP) in the manufacture of a product comprising a liquid composition for treating or preventing dentin hypersensitivity in a subject in need thereof, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP) stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP) stabilized amorphous fluorinated calcium phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, The liquid composition is mixed with another composition having an alkaline pH to form a mixed composition having a pH equal to or greater than 9, and the mixed composition is then applied to the exposed dentinal tubules to form a gel in and / or on the exposed dentinal tubules.

36. Use of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP) in the manufacture of a product, the product comprising a liquid composition for forming a protective layer on a tooth surface, the liquid composition comprising at least 40% w / v and less than 80% w / v of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), the pH of the liquid composition being less than or equal to pH 6, The liquid composition is mixed with another composition having an alkaline pH to form a mixed composition having a pH equal to or greater than 9, and then the mixed composition is applied to the tooth surface to form a protective layer on the tooth surface.

37. Use according to any one of claims 34 to 36, wherein the product is a cosmetic product.

38. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH have a pH of 8.

39. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH have a pH of 9.

40. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH have a pH of 10.

41. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH had a pH of 11.

42. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH have a pH of 12.

43. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH had a pH of 13.

44. The use according to any one of claims 34 to 36, wherein The other compositions having an alkaline pH had a pH of 14.

45. The use according to any one of claims 24 to 27 and 34 to 36, wherein The liquid composition also contains free fluoride ions.

46. ​​The use according to claim 45, wherein The free fluoride ions are present in the liquid composition at a concentration of 200 ppm to 10000 ppm.

47. The use according to claim 45, wherein The free fluoride ions are present in the liquid composition at a concentration of 2600 ppm to 8500 ppm.

48. The use according to claim 45, wherein The free fluoride ions were present in the liquid composition at a concentration of 8200 ppm.

49. The use according to any one of claims 24 to 27, wherein When the liquid composition is applied to a tooth surface or subsurface or lesion, the tooth surface or subsurface or lesion is heated.

50. The use according to any one of claims 24 to 27, wherein After the liquid composition is applied to a tooth surface or subsurface or lesion, the tooth surface or subsurface or lesion is heated.

51. The use according to any one of claims 24 to 27 and 34 to 36, wherein When the pH of the liquid composition applied to a tooth surface or subsurface or lesion is subsequently raised to a pH equal to or greater than 9, the tooth surface or subsurface or lesion is heated.

52. The use according to any one of claims 24 to 27 and 34 to 36, wherein After the pH of the liquid composition applied to the tooth surface or subsurface or lesion is subsequently raised to a pH equal to or greater than 9, the tooth surface or subsurface or lesion is heated.

53. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than 37°C.

54. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 40°C.

55. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 45°C.

56. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 50°C.

57. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 55°C.

58. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 60°C.

59. The use according to claim 52, wherein The tooth surface or subsurface or lesion is heated to a temperature greater than or equal to 65°C.

60. The use according to claim 52, wherein The tooth surface or sub-surface or lesion is heated to a temperature of less than 65°C.

61. The use according to any one of claims 24 to 27 and 34 to 36, wherein The liquid composition comprises more than 45% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

62. The use according to claim 61, wherein The liquid composition comprises more than 50% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

63. The use according to claim 61, wherein The liquid composition comprises more than 55% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

64. The use according to claim 61, wherein The liquid composition comprises more than 60% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

65. The use according to claim 61, wherein The liquid composition comprises 65% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

66. The use according to claim 61, wherein The liquid composition comprises 70% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

67. The use according to claim 61, wherein The liquid composition comprises 75% w / v and less than 80% w / v of stabilized ACP and / or stabilized ACFP.

68. The use according to any one of claims 24 to 27, wherein Prior to raising the pH of the liquid composition applied to the hypomineralized dental surface or subsurface or lesion to a pH equal to or greater than 9, the hypomineralized dental surface or subsurface or lesion is contacted with the liquid composition for up to 20 minutes.

69. The use according to claim 68, wherein In (i) the hypomineralized tooth surface or subsurface or lesion is contacted with the liquid composition for at least a few seconds to 5 minutes.

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

71. The use according to any one of claims 24 to 27 and 34 to 36, wherein Prior to contacting with the liquid composition, the hypomineralized tooth surface, subsurface or lesion has been acid etched.

72. The use according to any one of claims 1 to 17, 24 to 27 and 34 to 36, wherein The phosphopeptide is casein phosphopeptide.

73. The use according to any one of claims 1 to 17, 24 to 27 and 34 to 36, wherein The liquid composition, the other composition and / or the mixed composition is applied to a tooth surface, sub-surface or lesion by a dental health professional.

74. The use according to claim 73, wherein The liquid composition, the other composition and / or the mixed composition is applied to the tooth surface, sub-surface or lesion using a microbrush.

75. The use according to any one of claims 1 to 17, 24 to 27 and 34 to 36, wherein The subject of the identification has white spot lesions, fluorosis lesions, caries lesions or lesions caused by tooth erosion.

76. A kit for reducing the visibility of a hypomineralized tooth surface or subsurface, the kit comprising or consisting of: (a) a liquid composition comprising at least 40 wt % and less than 80 wt % of phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) and / or phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP), wherein the pH of the liquid composition is less than or equal to pH 6, and (b) Other compositions having an alkaline pH.

77. The kit according to claim 76, wherein: The kit further comprises instructions for use according to any one of claims 1 to 17, 24 to 27 and 34 to 36.

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